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 FUN Poster Session and Social Events and Abstract book Monday, November 17, 2014 6:45-­‐8:45 PM The Renaissance Washington, DC Grand Ballroom Welcome to the 2014 FUN Poster Session and Social – an event that embodies the mission of FUN to facilitate and celebrate undergraduate neuroscience research and education. This year is a record-­‐breaker, with over 200 posters being presented! We are tremendously thankful for the support of the Society for Neuroscience. In addition, we thank those institutions, organizations, and companies that have sponsored student travel awards (see below). Their commitment to undergraduate neuroscience research and education is greatly appreciated! Agenda for the Fun Poster Session and Social 6:45-­‐8:00 pm Poster presentations 8:00-­‐8:45 pm Awards ceremony Opening remarks Announcement of new officers Travel Award winners recognized Brain Awareness award winner recognized JUNE Editor Awards FUN Faculty Awards Mentor Award Educator of the Year Award Service Award Career Achievement Award Lifetime Achievement Award Recognition of the Out-­‐Going President Closing Remarks FUN Student Travel Awards Committee: Lisa Gabel (Lafayette College), Chair, President-­‐Elect Reviewers: Amanda Clinton, Dorothy Kozlowski, Elizabeth Meyer-­‐Bernstein, Katherine Mickley-­‐Steinmetz, Tracie Paine, Charles Weaver, Amy Beadles-­‐Bohling, Lora Becker, Greg Butcher, Aaron Cecala, Lee Coates, Sarita Lagalwar, Jacqueline Morris, Mary Morrison, James Roberts, Josef Trapani Travel Award Sponsors: Grass Foundation Med Associates Nu Rho Psi MBF Bioscience ADInstruments Carnegie Mellon University Leica Microsystems Faculty for Undergraduate Neuroscience Sinauer Associates Data Sciences International Noldus Information Technology San Diego Instruments Wellesley College Coulbourn Instruments, Panlab, & Harvard Apparatus Lafayette Instruments and Campden Instruments FUN Faculty Awards Committee: Noah Sandstrom (Williams College), Chair, Past-­‐President Jeffrey Smith (Saginaw Valley State University), President Elizabeth Meyer-­‐Bernstein (College of Charleston) Adrienne Betz (Quinnipiac University) 2 1 TRAVEL AWARD sponsored by Carnegie Mellon University Craig Russo Merritt Taylor Characterization of candidate genes that regulate stem cell differentiation in the developing chick embryo C.A RUSSO, K.M. MCKAY, K.A. MCGUIRE, M.K. TAYLOR Grand Valley State University The characterization of candidate regulators of stem cell differentiation can be performed in the chick embryo using quantitative PCR and anatomical techniques. Here we focus on one gene of interest, Barx2, on neural progenitor differentiation in the developing chick neural tube. Our work focuses on characterizing the localization of the endogenous protein in the developing neural tube and determining the effect of Barx2 overexpression. Email: [email protected] 2 TRAVEL AWARD sponsored by Leica Microsystems Daniel VanDerhoef Michelle Mynlieff Colocalization of interneuron markers with proteins involved in GABAB receptor mediated calcium current enhancement in neonatal hippocampus D.P. VANDERHOEF, M. MYNLIEFF Marquette University Previously, we demonstrated that activation of the G-­‐protein coupled GABAB receptor (GABABR) modulates currents through voltage-­‐gated calcium channels with an attenuation of N-­‐Type calcium current mediated by a Gαi/o G protein. Our laboratory has also described enhancement of L-­‐type calcium current mediated by a Gαq G protein and the α isoform of protein kinase C (PKCα). This enhancement peaks at postnatal day 7 and is expressed in only 10-­‐20% of cells in the superior region of the hippocampus. Since a large percentage of the cells in the CA1 region are excitatory pyramidal cells, we hypothesized that enhancement of current is confined to one or more inhibitory interneuron subtypes and determining the subtype may provide insight into a physiological role for the L-­‐type current enhancement observed. Studies have reported up to 21 different inhibitory interneuron subtypes. Sloviter et al. (Neuropharm., 38(11), 1707-­‐21, 1999) has demonstrated that in the CA1 region, interneurons expressing the neurochemical markers cholecystokinin (CCK), calbindin, neuropeptide Y, and somatostatin all express GABABR; making them possible candidates for the neuron(s) in which GABABR mediated calcium current enhancement occurs. Interneurons that encompass these parameters are the CCK+ basket cells, Schaffer collateral associated cells, and quadrilaminar cells. In the present study, fluorescent confocal microscopy in the superior hippocampal region from 6-­‐8 day old rats is utilized to determine colocalization of neurochemical markers used to identify interneuron subtypes with different components of the signaling pathway mediating calcium current enhancement. We hypothesized that the neuron of interest is a CCK+ basket cell, due to the morphology of the cell and similar localization as seen in preliminary data. The components of the signaling pathway of interest include GABABR, Gαq, PKCα and the two L-­‐type calcium channel isoforms expressed in the brain, CaV1.2 and CaV1.3. The specific isoform of L-­‐type channel enhanced by GABABR activation has not been determined but the developmental expression and colocalization with GABAB receptors suggest that CaV1.2 is a component in the pathway. Preliminary data demonstrate a much higher colocalization of calbindin with CaV1.2 than CCK with CaV1.2. However, only 1.17% of the total cells counted in the CA1 region express both calbindin and CaV1.2 and thus, it is likely that enhancement of the calcium current by GABAB receptor activation occurs in multiple interneuron subtypes. 3 3 TRAVEL AWARD sponsored by San Diego Instruments Jacob Stafford Ryan Bachtell Effects of adolescent caffeine consumption on anxiety behaviors, plasma corticosterone, and neural activity J. STAFFORD, C.E. O’NEILL, R.J. NEWSOM, S.C. LEVIS, T. SCOTT, R.K. BACHTELL University of Colorado at Boulder Caffeine is the most commonly used psychoactive substance worldwide, and consumption by children and adolescents has risen dramatically in recent years. Previous studies have found that energy drink use is associated with anxiety young adult males, and can induce panic attacks in patients diagnosed with panic disorder. These experiments examine the effects of adolescent caffeine consumption on anxiety related behaviors, basal and stress-­‐induced plasma corticosterone (CORT), and stress-­‐induced c-­‐fos mRNA expression. Beginning on post-­‐natal day 28 (P28), Sprague-­‐Dawley rats consumed caffeine (0.3 g/L) for 28 days (P28-­‐55). Age-­‐matched control rats consumed water. Caffeine and water consumption were monitored throughout the procedure, with no significant differences between groups on fluid consumption or body weight gain. Following 28 days of caffeine consumption, the caffeine solution was replaced with water for the remainder of the experiment. Behavioral testing occurred at three different time points: during caffeine exposure (P52-­‐55), short-­‐term withdrawal (24 hrs) from caffeine (P56), and long-­‐term withdrawal (1 week) from caffeine (P62-­‐66). Animals exposed to caffeine during adolescence exhibited increased anxiety in the open field at both withdrawal time points, 24 hrs and 1 week post caffeine, but not during the caffeine exposure. Chronic adolescent caffeine consumption also decreased social interaction with age-­‐matched conspecifics at both short-­‐ (24 hr) and long-­‐term withdrawal (1 week), but not during caffeine exposure. Finally, caffeine-­‐exposed rats demonstrated increased anxiety on an elevated plus maze at all three time points. Because systemic caffeine administration has been shown to increase plasma CORT levels, we examined the effect of adolescent caffeine consumption on basal and stress-­‐induced CORT levels during caffeine withdrawal. Animals exposed to caffeine had increased plasma CORT at the circadian trough, however no differences were observed at the circadian peak. Stress-­‐induced CORT was measured 30 min after animals were placed on a small pedestal elevated 2 feet off the ground for 5 minutes. Stress-­‐induced CORT was significantly lower in animals exposed to caffeine during adolescence. Together these findings suggest that adolescent caffeine consumption alters emotional reactivity using behavioral and neuroendocrine measures. We are currently processing the tissue to examine stress-­‐induced c-­‐fos mRNA expression in several stress-­‐responsive brain areas including the frontal cortex, the extended amygdala, lateral septum, and hypothalamic regions. Email: [email protected] 4 TRAVEL AWARD sponsored by the Grass Foundation Alexandria White Kyle J. Frantz Arc and BDNF expression after cocaine self-­‐administration or cue-­‐induced reinstatement of cocaine-­‐seeking in adolescent and adult male rats A.C. WHITE, C. LI, B.F. WILLIAMS, J. MCGINTY, K.J. FRANTZ Georgia State University In our laboratory, male rats that self-­‐administer cocaine as adolescents exhibit lower levels of cue-­‐induced reinstatement of cocaine-­‐seeking, as compared with adults. Activity-­‐regulated cytoskeletal-­‐associated gene (arc) and brain-­‐derived neurotrophic factor (bdnf) are two important neuroplasticity-­‐related genes that influence drug-­‐seeking behavior, change over development, and may play a role in age differences we observe. For the present study, we predicted that levels of cocaine-­‐seeking correlate negatively with differential expression of Arc and BDNF in reward and reinforcement-­‐related brain regions. Adolescent and adult rats were allowed to acquire lever-­‐pressing maintained by i.v. infusions of cocaine in daily two-­‐
4 hour sessions over 13 days. A subset of rats in both age groups received only saline infusions. At three experimental time points (immediately after the last self-­‐administration session, after extinction and reinstatement at 1 day abstinence from cocaine, and after extinction and reinstatement at 60 days abstinence), rats were sacrificed and brain tissue was collected. Arc and BDNF mRNA levels were analyzed by in situ hybridization and densitometry in the prelimbic and infralimbic cortex, nucleus accumbens core and shell, claustrum, caudate putamen, and motor cortex. Although Arc expression varied by drug treatment and time in region-­‐dependent ways, Arc expression was similar across age groups in almost all cases. In contrast, BDNF expression was higher in adolescent compared to adult rats. Additionally, BDNF expression was higher in cocaine-­‐experienced rats and rats sacrificed at the first and last experimental time points. These data generally support the hypothesis that higher levels of BDNF mRNA in reward and reinforcement-­‐related brain regions during adolescence could attenuate some long-­‐term effects of cocaine. Future studies should entail mechanistic analysis of BDNF mRNA and protein, as well as the role of BDNF receptors in age-­‐dependent cocaine-­‐related behaviors. Email: [email protected] 5 TRAVEL AWARD sponsored by Wellesley College Emily Powers Jean Hardwick Expression and actions of AT1, AT2 and Mas receptors in the guinea pig intrinsic cardiac plexus E.N. POWERS, K. A. LUCKETT, S.A. ROSEN, E.M. SOUTHERLAND, J.L. ARDELL, J.C. HARDWICK Ithaca College, East Tennessee State University The intrinsic cardiac (IC) nervous system integrates inputs from multiple sources to coordinate cardiac function. These inputs include parasympathetic efferent, sympathetic efferent and sensory afferent fibers in addition to local hormonal factors. A major local factor is angiotensin II (Ang II) which is found in the blood and is also produced by proteases within the cardiac interstitium. Ang II acts via AT1 or AT2 receptors. In addition, Ang II can be cleaved by ACE2 to formed Ang(1-­‐7), a peptide that has been found to have significant effects on neurons via activation of Mas receptors. In this study, we looked at the expression of the different angiotensin peptide receptors and their actions on neurons of the guinea pig intrinsic cardiac plexus. All three receptors, AT1R, AT2R, and MasR, were detected in homogenates of the isolated cardiac ganglion by Western blot analysis. Previous studies showed that AT2R can increase the neuronal responses to adrenergic and muscarinic signals. Using a whole mount in vitro preparation of the cardiac plexus, intracellular voltage recordings were made from individual IC neurons. We found that Ang(1-­‐7) synergistically increases neuronal excitability responses to NE and muscarinic agonists. We also examined the effects of Ang II on synaptic function in the ganglion by stimulating intraganglionic fibers leading to IC neurons of interest. Application of Ang II reduced the synaptic efficacy, as indicated by a reduced number of postsynaptic action potentials generated with increasing frequency suprathreshold stimulation. Addition of the AT1R antagonist losartan inhibited this effect. To examine potential changes in AngII receptor function with chronic heart disease, myocardial infarction (MI) was surgically induced in animals, and the cardiac ganglion examined 8 weeks later. Western blot analysis showed a decrease in AT1R expression in ganglia from MI animals, with no significant change in either AT2R or MasR expression levels. However, prior functional studies of the effects of Ang II on IC neurons demonstrated a MI-­‐induced loss in sensitivity to exogenous Ang II application. Combined, these results indicate that MI induces a down regulation in presynaptic AT1 receptors, as well as a decrease in signaling via AT2R. Thus, increases in Ang II production, such as occurs with chronic heart disease, can result in significant alterations in function within this ganglion. Grant/Other Support: : NIH HL98589, NIH HL71830. Email: [email protected] 5 6 TRAVEL AWARD sponsored by Sinauer Associates and FUN Nick Upright Julio J. Ramirez Characterization of the heterosynaptic interaction of the septal and crossed entorhinal projections to the dentate gyrus after unilateral entorhinal cortex lesion in rats: A time course study N.A. UPRIGHT, E.L. KRAUSE, G.R. SMITH, M.K. MOSES-­‐HAMPTON, P.G. LAKHMANI, J.J. RAMIREZ Davidson College Axonal sprouting has been investigated following various forms of damage to the CNS, including Alzheimer's disease, stroke, and traumatic brain injury. Alzheimer's disease is known to involve a marked degeneration of the entorhinal cortex (EC). A significant number of entorhinal projections to the hippocampus terminate in the ipsilateral dentate gyrus (DG); however, a small number of fibers project to the contralateral DG forming the crossed temporodentate (CTD) pathway. Following a unilateral EC lesion in rats, the glutamatergic CTD and the acetylcholinesterase-­‐containing, cholinergic septal input to the DG, termed the septodentate (SD) pathway, have been shown to undergo axonal sprouting. Lesion-­‐induced, CTD sprouting results in greater synaptic efficacy as early as 6 days postlesion relative to the normal CTD. The present study explored whether, relative to CTD stimulation alone, paired stimulation of the SD pathway followed by CTD stimulation would affect the response of the DG at 6, 8, 12, or 90 days postlesion. Male, Sprague-­‐Dawley rats were given either unilateral entorhinal lesions or sham operations, which consisted of a craniotomy over the entorhinal area. Stimulating electrodes were placed in the medial septum and in the intact, contralateral EC 6, 8, 12, or 90 days after a unilateral lesion or sham operation. Evoked, field excitatory postsynaptic potentials (fEPSPs) were recorded in the DG ipsilateral to the lesioned EC. The paired-­‐pulse paradigm involves stimulation of an initial input, termed the "conditioning pulse," followed by a second input, termed the "test pulse." In the heterosynaptic paired-­‐
pulse stimulation portion of this study, SD stimulation preceded CTD stimulation at a range of interpulse intervals (IPIs; from 30 to 500 ms). Relative to control cases, unilateral EC lesions significantly increased the fEPSPs produced by CTD stimulation alone at all the time points examined. In contrast, compared to CTD stimulation alone, pairing SD with CTD stimulation significantly depressed the amplitude of the fEPSPs across IPIs at 8, 12, and 90 days postlesion. Stimulation of the CTD alone rarely produced granule cell discharge (i.e., population spikes) at the time points we explored; however, paired stimulation of the SD and CTD produced granule cell discharge in a significant number of cases at the 90-­‐day time point with a response profile similar to that observed after paired stimulation of the SD and the perforant path in intact cases. Thus, heterosynaptic stimulation in which the septal input and crossed entorhinal input are paired following entorhinal lesions significantly affects DG responsivity. This research was made possible through support from the National Science Foundation and the National Institutes of Health. Email: [email protected] 7 TRAVEL AWARD sponsored by FUN Mackenzie Thibault Alfred J Robison Region-­‐specific induction of FosB isoforms in mouse brain after stress or chronic fluoxetine exposure M.A. THIBAULT, A.L. EAGLE, S. KASKA, E.J. NESTLER, M.S. MAZEI-­‐ROBISON, V. VIALOU, AND A.J. ROBISON Michigan State University An estimated 1 in 10 U.S. adults report depression, but current pharmacological therapies are effective in only about 50% of patients. In order to uncover novel gene targets that may allow for therapeutic treatment of currently refractive individuals, we examine the gene targets of transcriptional changes that occur with chronic exposure to antidepressants, including the selective serotonin reuptake inhibitor 6 fluoxetine. We have previously shown that induction of the transcription factor ΔFosB in the nucleus accumbens (NAc) of mice promotes resilience to the social defeat model of depression and is required for fluoxetine-­‐mediated reversal of the social defeat phenotype (Vialou et al., 2010). Here, we show by immunohistochemistry that FosB isoforms are induced in more than 25 different brain regions by chronic fluoxetine, including many regions classically associated with depression and mood, i.e. hippocampus (HPC) and prefrontal cortex (PFC). Further, we demonstrate by Western blot that FosB isoforms are differentially expressed by brain region, both in the basal state and after fluoxetine-­‐mediated induction. While HPC and PFC have low basal levels of FosB isoforms and show high levels of ΔFosB after fluoxetine, NAc has a higher basal expression of FosB, ΔFosB, and Δ2ΔFosB, and shows a proportionally greater induction of full-­‐length FosB after fluoxetine. We also observe differential isoform expression in these brain regions after chronic social defeat stress, indicating that FosB isoform expression patterns may underlie susceptibility and resilience to stress. Because each FosB isoform has different gene targets that may very further by brain region, we hypothesize that determining the brain-­‐region-­‐specific gene expression changes mediated by FosB isoforms may uncover novel targets for therapeutic intervention in mood disorders that could improve treatment of currently refractive individuals. In line with this hypothesis, we are currently conducting experiments to determine whether ΔFosB expression in hippocampus controls mood or mediates the antidepressant effects of fluoxetine and are exploring ΔFosB gene targets in this and other brain regions. This work was supported by grants from NIDA, NIMH, and the Whitehall Foundation. Email: [email protected] 8 TRAVEL AWARD sponsored by Lafayette Instruments and Campden Instruments Megan Kechner Michelle Mazei-­‐Robison Physical and psychological stress increase voluntary morphine consumption M. KECHNER, S. KASKA, S. COOPER, M.S. MAZEI-­‐ROBISON Michigan State University Stress exposure is known to influence drug craving and relapse in human subjects and these effects can be modeled in preclinical rodent models. Social defeat stress (SDS), an ethologically valid model that utilizes the physical and psychological stress imposed by social subordination, has been demonstrated to exhibit excellent face and pharmacological validity for stress-­‐related disorders like depression. The social defeat model has been used to assess drug behavior, with results generally supporting increased drug reward and self-­‐administration following SDS. However, SDS, as well as most other rodent models of mood disorders, utilize some form of physical trauma, complicating the study of pain-­‐relieving opiate drugs. To overcome this, we utilized the recently developed model of emotional stress. In this model, mice witness, but are not physically exposed to, chronic SDS (CSDS). Given the co-­‐morbidity of stress-­‐related disorders and opiate dependence, we sought to examine whether physical or emotional defeat stress increased morphine reward and consumption. For physical CSDS, we utilized the previously validated paradigm and emotional stress was completed as described by Warren et al. (2013). To assess voluntary morphine consumption, we used a two-­‐bottle choice paradigm that takes advantage of the genetic background of C57BL6 mice, which have been shown to exhibit a propensity for morphine drinking. After establishing a morphine concentration that resulted in an ~75% preference for morphine in control mice, we assessed morphine consumption following physical and emotional stress. We found that both physical and emotional stress significantly increased morphine preference in the two-­‐bottle choice assay. We found that there was a significant negative correlation between SI ratio and morphine preference, where decreasing SI ratio correlates with an increase in morphine reward. Surprisingly, morphine preference did not differ between the physical and emotional stress groups, even though the emotional stress mice exhibited modest social avoidance at this time-­‐point. We are currently evaluating whether this increase in morphine preference persists at longer time-­‐points. These data suggest that both physical and emotional social defeat stress increase morphine reward and these studies, along with others to examine the molecular mechanisms underlying susceptibility to emotional stress, offer promise of an increased understanding of the neurobiological mechanisms that contribute to opiate use. 7 Email: [email protected] 9 TRAVEL AWARD sponsored by FUN Renee Rotolo Adrienne Betz Chronic oral riluzole or caloric restriction ameliorate symptoms of experimental autoimmune encephalomyelitis R. ROTOLO, J. DEMURO, G. DRUMMOND, J. WOOD, E. LAZAROFF, S. LUPINSKI, C. LITTLE, A. WOLF, G. VANN, L. TELISKA, D. RILEY, J. BAHGAT, J. VIDAL, M. ALBALAWI, L. JOHNS, A. BETZ Quinnipiac University Experimental Autoimmune Encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS). We characterized the impairments that represent EAE in female C57BL/6 mice. Mice were immunized subcutaneously with 100 μg of myelin oligodendrocyte glycoprotein emulsified in incomplete Freund's adjuvant supplemented with 500ug mycobacterium tuberculosis H37RA and 200 ng of intraperitoneal pertussis toxin on days 0 and 2. Tail paralysis was observed daily. In Experiment 1, we found that a battery of behavioral tasks delayed the onset and severity of EAE but did not affect nociception. In Experiment 2, we found that caloric restriction (CR) and chronic oral administration of riluzole, a glutamate antagonist, delayed the onset and severity of EAE. Additionally, CR and riluzole both reduced nociceptive behavior throughout disease progression. Glutamate neurotoxicity has been proposed as major determining factor that accompanies the demyelination and axonal degeneration observed during the course of MS. Further, CR plus riluzole decreased symptomology more so than CR alone. Lastly, we found altered levels in proteins important for normal immune reactions such as TNFα, Treg, IL-­‐6, BDNF, pSTAT3, and leptin. Altered immunological function was also indicated by reduced demyelination in the spinal cords of mice treated with riluzole. These findings indicate a compelling need to delineate the roles of glutamate, the immune response, and CR in EAE. Email: [email protected] 10 TRAVEL AWARD sponsored by Nu Rho Psi Lucas Huffman Gary L. Dunbar Bone marrow-­‐derived mesenchymal stem cells in the suppression of highly proliferative glioblastoma multiforme L.D. HUFFMAN, D.J. DUES, A. CRANE, K.R. IDYLE, E.S. DWENGER, K.D. FINK, J. ROSSIGNOL, G.L. DUNBAR Central Michigan University Glioblastoma multiforme (GBM) represents the most aggressive and infiltrative primary tumor formation of the central nervous system (CNS). Current treatments have minimal therapeutic efficacy, establishing a substantial need for novel therapeutic strategies to improve patient outcome. Bone marrow-­‐derived mesenchymal stem cells (MSCs) offer an innovative approach in the treatment of GBM. Previous experimentation in our lab has demonstrated that MSCs exhibit a significant suppressive effect on glioma tumor cell growth in vitro. However, given the insufficiency of information regarding the effectiveness of treating GBM with MSCs, the goal of this study was to assess the effects and outcome of transplanted MSCs on tumor proliferation in vivo using an aggressive F98 glioma cell model of GBM. Three groups of 2 month-­‐old Sprague Dawley rats (n=8) were used. SHAM controls received only vehicle treatments (Hank’s Buffered Salt Solution) rather than either the F98 cells that produce GBM or the MSCs used to treat the GBM. Another group received only F98 cells (producing GBM) and the other received F98 cells, followed by injections of MSCs as a potential treatment. The rats activity levels were monitored in an open-­‐field test and their general health and length of survival recorded throughout the study. In addition to statistical 8 comparisons of activity levels, general health measures, and survival, immunohistological comparisons of tumor size and immune response were assessed. Our results suggest that MSC treatments may increase survivability and improve behavioral outcomes by suppressing tumor proliferation and decreasing overall tumor volume. Email: [email protected] 11 TRAVEL AWARD sponsored by Nu Rho Psi Sarah Cottrell-­‐Cumber Alev Erisir Multiple sclerosis therapeutic glatiramer acetate enhances oligodendrocyte precursor cell differentiation and remyelination. S. COTTRELL-­‐CUMBER, A. ROSEN, A. FERNANDEZ-­‐CASTANEDA, A. GAULTIER University of Virginia Multiple Sclerosis (MS) is a progressive neurological illness characterized by destruction of the myelin sheath surrounding neurons in the central nervous system. The destruction of the myelin sheath is mediated by autoreactive T cells. Current pharmacological therapies seek to decrease the severity of the inflammatory response on the damaged axons by suppressing various aspects of the immune system. One such current MS treatment, Glatiramer Acetate (GA), is believed to act as an immunomodulatory peptide that shifts the immune response by simulating myelin basic protein. However, evidence suggests that GA also has the ability to directly modulate oligodendrocyte progenitor cells function (OPCs) to enhance differentiation and remyelination in MS lesions. Chronic demyelination is a major cause of neurodegeneration in MS patients. The CNS contains OPCs that have the potential to differentiate into mature oligodendrocytes and remyelinate denuded axons. However, myelin debris lingering in the MS plaques inhibits the process of axon remyelination. Here we demonstrate an immune independent action of GA and OPCs, through the enhancement of OPC differentiation and remyelination. Our data shows that GA increases markers of myelination in CG4 cells, an OPC cell line. We have observed increased myelin basic protein (MBP) RNA and protein expression in CG4 cells treated with GA. Since myelin is known to inhibit the differentiation of OPCs, we will challenge OPCs with myelin, and then measure production of MBP in the presence or absence of GA. We expect to see increased MBP expression when GA is applied; this would indicate the ability of GA to overcome myelin inhibition and augment remyelination. We believe GA promotes OPC differentiation in MS, which could explain its beneficial effects as a therapeutic; further identification of a downstream signaling pathway could lead to the development of a novel remyelination therapeutic. Research supported by The University of Virginia Center for Undergraduate Excellence Harrison Research Award and The University of Virginia College Council Minerva Award Email: [email protected] 12 TRAVEL AWARD sponsored by Sinauer Associates Katherine Harmon Patsy S. Dickinson Mechanisms and effects of stretch feedback in the heart of the American lobster, Homarus americanus K. HARMON, M. CHIN-­‐PURCELL, E.S. DICKINSON, T.M. HARTLEY, O. ELLERS, A.S. JOHNSON, P.S. DICKINSON Bowdoin College Although central pattern generators (CPGs) can produce rhythmic outputs in isolation, their outputs in vivo are often altered by both sensory feedback and neuromodulators. The interactions of these components are not well understood. We examined the neurogenic heart of the lobster, Homarus americanus, which is controlled by a small CPG, the cardiac ganglion (CG). Previous research suggests that 9 crustacean heart muscle provides feedback to the CG about the degree of cardiac filling, mediated by stretch sensitive dendrites emanating from the CG neurons. We thus investigated the effects of stretch on CG output and the mechanisms underlying stretch sensitivity. Dendrites of CG motor neurons were identified using intracellular dye fills. To determine the role of stretch-­‐sensitive dendrites, isolated hearts were stretched while heart contractions and CG motor output were recorded. Tonic stretches of the heart significantly increased contraction frequency in most preparations; the proportion that increased depended on direction of stretch. Frequency in the remaining preparations decreased or did not change. Removing stretch feedback either by severing the branches of the CG that are thought to contain the dendrites or by cutting the CG motor nerves decreased or eliminated the response to stretch, but increased the variability of the heartbeat. Additionally, both frequency and amplitude of heart contractions decreased slightly when the CG motor neuron dendrites were severed; frequency likewise decreased when the motor nerves were severed. Our data suggest that feedback is mediated largely by dendrites branching from the main CG trunk, and is triggered primarily by active muscle contractions. This feedback appears to stabilize the heartbeat and simultaneously enhance the activity of the CPG. To determine the cellular mechanism that underlies stretch sensitivity, we recorded intracellularly from CG motor neurons while manually stretching small bundles of muscle fibers. Stretch of a single muscle bundle did not alter cycle frequency, and we saw no changes in the membrane potential between bursts. Surprisingly, the amplitude of the driver potentials that underlie bursting in the CG neurons decreased when the attached muscle was stretched. Both the changes in heartbeat frequency in response to removal of the dendrites and the responses of the membrane potential to stretch differ considerably from those previously recorded in other species, suggesting a different mechanism underlying stretch sensitivity in this crustacean species. Experiments examining the effects of neuromodulators on this CPG-­‐effector system are underway. Support: NSF IOS-­‐1121973, NIH (INBRE) 5P20-­‐RR-­‐016463, 8P20-­‐GM-­‐103423 13 TRAVEL AWARD sponsored by Coulbourn Instruments David Swygart Matthew Kreitzer HCO3-­‐-­‐dependent, K+-­‐induced increase in proton flux at the endfoot of isolated muller cells of the tiger salamander D.I. SWYGART, R. KAUFMAN, B.K. TCHERNOOKOVA, B. WILLIAMS, M. OSBORN, B. SKINNER, E. NAYLOR, R.P. MALCHOW, M.A. KREITZER Indiana Wesleyan University Within the retina and the broader CNS, synaptic transmission is extremely sensitive to minute changes in pH. A growing number of studies suggest that regulation of extracellular pH plays an important role in shaping neuronal communication.. Tightly regulated levels of HCO3-­‐ are an understated contributor to extracellular pH. Levels of this extracellular pH buffer are impacted by blood flow, CO2 levels, as well as the expression and activity of HCO3-­‐ transporters and the enzyme carbonic anhydrase. Previous work (Newman, 1996) detected the presence of HCO3-­‐ transporters and carbonic anhydrase on radial glia (Müller cells) that span much of the overall thickness of the retina. The Müller cell plays a primary role in regulating many aspects of the retinal environment, such as ion, H+, and neurotransmitter levels, and an active role in the release of gliotransmitters. Newman’s observations suggested a HCO3-­‐-­‐dependent mechanism by which high extracellular K+ acidified the extracellular environment at their endfoot. This mechanism could be important for H+ clearance to the vitreal surface of the retina as well as in regulating neuronal communication during times of increased neuronal activity. Our findings, using a novel ultrasensitive H+-­‐selective self-­‐referencing system in combination with a newly develop CO2 chamber, corroborate these previous studies. The self-­‐referencing system utilizes a H+-­‐selective microelectrode that records measurements from a near and a far point from a cell in order to obtain a differential pH value 1000 times more sensitive than a stationary pH-­‐selective electrode. Our work suggests that Müller cells, isolated from tiger salamander retina, respond to increased K+o with an extracellular acidification at the endfoot. This acidification can be abolished when extracellular Na+ or HCO3-­‐ is removed or in the presence of the HCO3-­‐ transport antagonist, DIDS. This DIDS-­‐sensitive pH regulatory mechanism could also evoke 10 large extracellular alkalinizations and acidifications when the bathing media was changed between a low Na+ and normal Na+ environment, respectively. These findings extend previous work strongly implicating an important role for HCO3-­‐ in shaping extracellular pH by Müller cells in the retina. They warrant future studies to characterize whether these bicarbonate-­‐mediated alterations in pH contribution in a significant wayto the processing of visual signals in the retina. Email: [email protected] 14 TRAVEL AWARD sponsored by FUN Jessica Phan Kevin Sinchak Progesterone receptor-­‐B and Src kinase complex in the plasma membrane of the arcuate nucleus of the hypothalamus of female rats J. PHAN, M. MAHAVONGTRAKUL, K. SINCHAK California State University Long Beach Ovariectomized (OVX) rats treated with a 2μg dose of estradiol benzoate (EB) and subsequently given 500μg of progesterone 26 hours later are maximally sexually receptive four hours later. EB priming upregulates progesterone receptor-­‐B (PR-­‐B) necessary for lordosis. PR-­‐B is a classical nuclear transcription factor found in both the nucleus and cytoplasm. However, PR-­‐B has been localized to the plasma membrane by in vitro expression studies (Pedram et al., 2007; Welter et al., 2003) and in vivo in the arcuate nucleus of the hypothalamus (ARH; Mahavongtrakul et al., SFN, 2013). Progesterone has been shown to facilitate lordosis within 30 minutes, indicating rapid extranuclear PR signaling mechanisms are involved. This rapid progesterone facilitation of lordosis may be mediated through PR interactions with Src family kinase (Src). We have shown that Src activation in the ARH rapidly facilitates lordosis, and our behavioral data indicate that progesterone and Src signaling converge with dopamine receptor-­‐type 1 (D1) signaling to inhibit the output of ARH β-­‐endorphin neurons to facilitate lordosis. Although we have shown that PR and D1 do not complex, it is unclear whether a direct PR-­‐Src interaction occurs to mediate progesterone facilitation of lordosis. To determine whether Src complexes with PR-­‐B on the plasma membrane, OVX rats were treated with either oil or 2μg EB and 30 hours later plasma membrane fractions were extracted from ARH block dissections. Western blot analysis showed that PR-­‐B and Src are both present on the plasma membrane and co-­‐immunoprecipitation results indicate that PR-­‐B, but not PR-­‐A, is complexed with Src on the plasma membrane. This demonstrates the potential for direct PR-­‐B signaling through Src that is initiated at the plasma membrane level in the ARH to facilitate lordosis. We have also observed that Src levels do not change in ARH whole tissue preparations in both oil-­‐ and EB-­‐treated rats. However, it is unclear whether estradiol or progesterone treatment affects levels of PR, Src, and PR-­‐Src complexes on the plasma membrane. Thus, our results indicate that the rapid effects of progesterone may be mediated directly through PR-­‐Src signaling initiated at that the level of the plasma membrane to facilitate lordosis. Grant funding -­‐ R01HD058638NICHD NIH 15 TRAVEL AWARD sponsored by the Grass Foundation Ariel Lopez Carolyn Pytte Mold exposure alters adult new neuron migration in the mouse hippocampus A. LOPEZ, N. ABREU, K. PAGE, E. NORMAND, B. SHAUKAT, N. ADAMS, K. LIN, L. BICKERTON, C.F. HARDING, C.L. PYTTE CUNY Queens College Exposure to environmental mold is a growing concern for individuals living or working in water-­‐damaged buildings. In addition to a suite of somatic effects, mold exposure is also associated with cognitive impairments. Alarmingly, neurologists cannot differentiate between cognitive deficits in patients exposed 11 to mold and those with mild traumatic brain injury. Prior research in our lab has found that mold exposure leads to a decrease in hippocampal-­‐dependent memory in a contextual fear task. We have also demonstrated that mold exposure leads to a significant decrease in hippocampal new neuron survival assessed in mature (~35-­‐day old) neurons. Moreover, numbers of mature new neurons were negatively correlated with numbers of cells expressing the inflammatory interleukin-­‐1 beta (IL-­‐1β). Here we further characterize the effects of mold on hippocampal neurogenesis. To determine whether mold inhalation specifically targets the survival of mature neurons, we quantified numbers of younger hippocampal neurons (<30 days old) expressing doublecortin (DCX) in mice treated with mold. New neurons are primarily incorporated into the granular layer of the dentate gyrus; however, in damaged brains, ectopic neuron incorporation is seen, particularly in the hilar region. Therefore, we also assessed the relative numbers of mature new neurons seen in the granular layer and hilus. C57BL/6 mice were given intranasal instillations of 1) intact Stachybotrys spores (IN), 2) extracted Stachybotrys spores that had toxins removed and proteins denatured leaving skeletal elements (EX), or 3) saline vehicle (VEH). Mice were treated 3x per week for 6 weeks. We injected mice with bromodeoxyuridine (BrdU) 31-­‐37 days and 3 hours before sacrifice. Immunohistochemistry was used to visualize cells expressing BrdU and the neuronal marker Hu. We also labeled cells expressing doublecortin and the inflammatory marker, IL-­‐1β. We found that numbers of mature new neurons were negatively correlated with IL-­‐1β, whereas numbers of young neurons were positively correlated with IL-­‐1β. This suggests that inflammatory effects of mold exposure decrease new neuron survival specifically in older neurons (by age 31-­‐37). Increased numbers of young DCX-­‐expressing neurons may reflect compensation for cell death of older cohorts. In addition, mice exposed to mold spores had fewer new neurons in the granular layer and greater numbers of new neurons in the hilus than vehicle controls. These findings suggest that impaired hippocampal-­‐dependent learning may also be associated with aberrant new neuron migration in addition to decreased new neuron numbers. Email: [email protected] 16 TRAVEL AWARD sponsored by Noldus Information Technology Bobbie Stubbeman Mark E. Bardgett Early-­‐life risperidone administration enhances locomotor responses to amphetamine during adulthood B.L. STUBBEMAN, C.J. BROWN, M.E. BARDGETT Northern Kentucky University Antipsychotic drug prescriptions for pediatric populations have increased tremendously over the past 20 years, particularly the use of atypical antipsychotic drugs such as risperidone. In rats, forebrain dopamine receptor densities are elevated upon cessation of early-­‐life risperidone administration. This finding suggests that adult rats administered risperidone early in life should display enhanced behavioral sensitivity to drugs that elevate dopamine neurotransmission. This hypothesis was tested by measuring locomotor responses to amphetamine -­‐ a drug that releases forebrain dopamine – in adult rats administered risperidone early in life. Thirty-­‐five Long-­‐Evans rats received one of four doses of risperidone (vehicle, 0.3, 1.0, or 3.0 mg/kg) daily from postnatal day 14 through 42. Beginning on postnatal day 75, locomotor activity was recorded for 30 minutes once a week for four weeks. After 30 minutes, each rat received a subcutaneous injection of one of four doses of amphetamine (saline, 0.3, 1.0, or 3.0 mg/kg) in a counter-­‐balanced order across the four weeks. Locomotor activity was measured for 27 hours after amphetamine administration. Activity levels did not differ between the vehicle and risperidone groups for six hours after saline injection. Rats administered risperidone early in life displayed significantly greater locomotor activity for six hours after amphetamine injection. This effect was most prominent in the first two hours after injection of the 0.3 and 1.0 mg/kg amphetamine doses, and was seen in all groups administered risperidone early in life, but was most marked in the risperidone 3.0 mg/kg group. The results suggest that the development of forebrain dopamine systems is permanently altered by early-­‐life antipsychotic drug administration. The data raise concerns about possible increases 12 in sensitivity to recreational and therapeutic drugs that target dopamine in adults treated with antipsychotic drugs during childhood. Email: [email protected] 17 TRAVEL AWARD sponsored by Med Associates Inc. Kurt Fraser Shelly B. Flagel Stimulation of dopamine D3 receptors attenuates the expression of pavlovian conditioned approach responses and motivation for incentive cue presentation K.M. FRASER, J.L. HAIGHT, S.B. FLAGEL University of Michigan-­‐Ann Arbor Cues in the environment can guide behavior in adaptive ways, bringing one in close proximity to valuable resources (e.g. food, water, sex). However, some cues, specifically those that attain incentive salience, can gain inordinate control over behavior and direct actions in a maladaptive manner, as is evident in addiction and other disorders of impulse control. Reward cues acquire incentive salience via Pavlovian learning processes. Although a role for dopamine has been identified in these processes, it is still unclear as to which receptors are involved. Here we investigated the role of the dopamine D3 receptor in the attribution of incentive salience to discrete reward cues. To do this, we utilized an animal model that allows us to parse the neurobiological processes underlying the attribution of incentive vs. predictive value to a discrete cue paired with food-­‐delivery. Outbred Sprague-­‐Dawley rats were first characterized as sign-­‐ or goal-­‐trackers based on 7 days of Pavlovian conditioning wherein brief presentation of a lever-­‐cue was paired with delivery of a food reward. Sign-­‐trackers attribute incentive salience to the reward-­‐paired cue, as measured by interaction with the cue upon its presentation. In contrast, goal-­‐trackers use the reward-­‐paired cue merely as a predictor of reward delivery and upon its presentation orient behavior accordingly towards the food cup (i.e. the goal). Following acquisition of these respective conditioned responses, we used a within subject design to determine the role of the D3 receptor in the expression of sign-­‐ and goal-­‐tracking behaviors. Increasing doses (0.01-­‐0.32 mg/kg) of 7-­‐OH-­‐DPAT were administered and compared to the effects of vehicle injections on alternating days. We found that stimulation of the dopamine D3 receptor attenuates the expression of both of sign-­‐ and goal-­‐tracking behavior. There was a dose-­‐dependent effect on sign-­‐tracking behavior, with the largest effects in response to the two highest doses of drug. Interestingly, goal-­‐tracking was attenuated only at the lowest drug doses. To further examine the effects of D3 stimulation on the attribution of incentive salience to a discrete reward cue, we examined the effects of 7-­‐OH-­‐DPAT on the conditioned reinforcing properties of the lever-­‐cue. Administration of the D3 agonist significantly attenuated the motivation to work for presentation of the lever-­‐cue in sign-­‐trackers, but not goal-­‐trackers. This work highlights a role for the D3 receptor in the expression of Pavlovian conditioned approach responses and in the attribution of incentive salience to discrete reward cues. Email: [email protected] 18 TRAVEL AWARD sponsored by Lafayette Instruments and Campden Instruments Kerri Szolusha Michael Burman FAAH inhibitor OL-­‐135 disrupts contextual, but not auditory, fear conditioning in rats K. SZOLUSHA, R. BIND, K. KERNEY, D. BOGER, E. BILSKY, M. BURMAN University of New England Anxiety disorders are the most common psychological disorder, with an approximately 25% lifetime incidence. The endocannabinoid system has increasingly been the target of investigation for a potential role in fear and anxiety. Research on compounds such as CB1 agonists and antagonists have demonstrated 13 effects on fear and anxiety using both innate anxiety-­‐like behaviors and conditioned fear. However, direct CB1 agonists and antagonists often have undesirable cognitive side effects. Fatty acid amide hydrolase (FAAH) is a degradation enzyme targeting the endocannabiniods. Disrupting FAAH avoids many of the deleterious effects of direct CB1 manipulation. Interestingly, drugs that inhibit FAAH have two major effects: they enhance memory formation and they inhibit innate measures of anxiety. They don’t appear to have been assessed in classical fear conditioning, where these two effects would appear to compete. The current studies utilize classical fear conditioning, a particularly successful model of fear/anxiety, in which a previously neutral cue is associated with an aversive stimulus, such that the neutral cue comes to elicit a conditional fear response. In addition to fear for the explicitly conditioned cue (auditory fear), fear also develops to the conditioning apparatus and situation (contextual fear conditioning). These two tasks also require different neural substrates, in that fearing an auditory cue requires the amygdala and its efferent and afferent connections, whereas contextual fear conditioning recruits additional hippocampal and cortical circuitry. To examine the role that endocannabiniods play in fear conditioning, we injected the FAAH inhibitor OL-­‐135 at 5.6 mg/kg and 10 mg/kg both before and after fear conditioning. We then assessed both auditory and contextual fear. When the FAAH inhibitor was injected before conditioning there was a specific deficit in contextual fear. That auditory fear expression was intact and there was no difference in the shock reactivity of the animals suggests that these results are not due to analgesic or non-­‐
specific effects of the drug. When the animals were injected after training there was no effect, suggesting the endocannabinoid system is only required during acquisition of fear. These data are consistent with previous research showing that CB1 manipulation causes deficits of acquisition, but not consolidation, of fearful memories. This may further the promise of these compounds for the treatment of fear and anxiety. Email: [email protected] 19 TRAVEL AWARD sponsored by MBF Bioscience Johnny Huang Martin Wojtowicz Physical exercise prevents suppression of hippocampal neurogenesis and mitigates cognitive impairment in chemotherapy-­‐treated rats J. HUANG, G. WINOCUR, J.M. WOJTOWICZ University of Toronto Increasing evidence shows chemotherapy-­‐induced cognitive impairments in humans and rodents. These deficits include confusion, memory loss, decreased attention span, and inability to focus or concentrate. Physical exercise is known to enhance hippocampal neurogenesis and improve cognitive function. In this study we examined the effect of physical exercise on rats that were treated with chemotherapeutic agents. 3-­‐month-­‐old Long-­‐Evans rats (n=37), housed in either standard cages or cages that allowed unlimited access to a running wheel, received intra-­‐peritoneal injections of 5-­‐fluorouracil and methotrexate, or equal volumes of saline. They subsequently underwent a series of cognitive tasks -­‐ including spatial memory (SM), non-­‐matching-­‐to-­‐sample rule learning (NMTS), and delayed NMTS (DNMTS) tests. Expression levels of Ki67, doublecortin (DCX) were examined in the dentate gyrus. Analysis of variance (ANOVA) was used to test differences between groups on behavioral measures and cell counts. First, chemotherapy significantly reduced the number of DCX+ cells in the dentate gyrus by approximately 25% (P=0.02), while running markedly increased DCX+ cells (P<0.01). In addition, rats exposed to running showed evident increases in both Ki67+ (P<0.05) and BrdU+/DCX (P<0.05) cells. No significant difference in average running distance was observed between saline-­‐ and chemotherapy-­‐treated rats (P>0.25). Behaviorally, ANOVAs showed significant main effects of chemotherapy on performance in SM, NMTS and DNMTS. Running improved the performance on all these tasks. In summary, we report promising results where the adverse effects of chemotherapy on both hippocampal neurogenesis and behavioral performance were rescued by running, indicating physical exercise as a highly feasible and safe therapeutic intervention against chemotherapy-­‐induced cognitive deficits. Email: [email protected] 14 20 TRAVEL AWARD sponsored by Data Sciences International (DSI) Molly Barlow David C. Jewett Pramipexole decreases the discriminative stimulus effects produced by 22 hours food deprivation and food intake. M.A. BARLOW, K.A. TWAROSKI, H.M. BADZINSKI, K.J. OLSON, M.A. VANDEN AVOND, C.A. TODDES, B. GOMER, B. BERTI, A.R. JOHNSON, D.C. JEWETT University of Wisconsin -­‐ Eau Claire We have developed and refined a food-­‐deprivation discrimination paradigm that may serve as an animal model of ‘hunger’. We examined the ability of pramipexole, a D2/D3 agonist used clinically to treat Parkinson’s disease and restless leg syndrome, to reduce the effects of acute food deprivation in rats trained to discriminate between 2 and 22 hrs food deprivation in an operant choice paradigm. Generalization testing began after the acute food deprivation discrimination was acquired (~90 daily sessions). Prior to cumulative dose-­‐generalization tests, subjects were food deprived for 22 hrs. Injections of vehicle and pramipexole (0.001-­‐0.032 mg/kg, s.c.) occurred every 35 minutes (a 30 min pretreatment time and a 5 min response period) until a complete pramipexole dose-­‐effect function was generated. Food intake was measured for 1 hr after the generalization tests. Pramipexole (0.01 mg/kg) significantly reduced the discriminative stimulus effects of 22 hrs deprivation. Pramipexole (0.01-­‐0.032 mg/kg) also significantly reduced response rates and post-­‐session food consumption. These results are consistent with our previous research demonstrating amphetamine decreases the discriminative stimulus effects of 22 hrs food deprivation and support the hypothesis that dopamine mediates food consumption by mechanisms related to ‘hunger.’ University of Wisconsin -­‐ Eau Claire Faculty/Student Research Collaboration grant 21 TRAVEL AWARD sponsored by Nu Rho Psi Veronica Burnham Jan E. Thornton Blocking luteinizing hormone but not gonadotropin-­‐releasing hormone in the dorsal hippocampus rescues spatial memory deficits in ovariectomized female rats. V.L. BURNHAM, A GOLDBERG, J.E. THORNTON Oberlin College Recent work has implicated the activity of luteinizing hormone (LH) in post-­‐menopausal memory decline. In rodents, increasing physiological levels of LH has been shown to significantly decrease performance on spatial memory tasks. Previous research has also indicated that blockage of gonadotropin releasing hormone (GnRH) activity, which decreases LH levels, can cause amelioration of spatial memory deficits. However, it is unclear whether directly reducing activity of LH in the brain is able to rescue spatial memory deficits in individuals with high LH. To further investigate the role of hippocampal LH receptors in spatial memory deficits, female Sprague-­‐Dawley rats were ovariectomized (ovx) and implanted with either an estradiol (E) or blank (blk) capsule. Bilateral cannulae were implanted into the dorsal hippocampus (DH). Animals received infusions of 0.9% saline (vehicle), the LH homologue hCG (human chorionic gonadotropin), or deglycosylated-­‐hCG (dg-­‐hCG; an LH receptor antagonist), 3-­‐5 hours prior to behavioral testing via Object Location Test (OLT). Consistent with previous results, estradiol enhanced spatial memory (ovx + E compared to ovx) and ovx + E animals receiving hCG infusions into the dorsal hippocampus showed a significant decrease in spatial memory compared to vehicle infusions. Importantly, infusion of the LH receptor antagonist dg-­‐hCG into the dorsal hippocampus of ovx + blk animals caused a rescue of spatial memory deficits induced by ovariectomy. These data indicate that LH acts on the hippocampus to modulate spatial memory. Whether GnRH antagonism exerts its effects on spatial memory via its effects on LH or thru some more direct action on the dorsal hippocampus was also 15 assessed. In the present experiments infusion of Antide via bilateral cannulae into the DH of ovx + E and ovx + blk animals 4-­‐6 hours prior to administration of behavioral tests did not result in any significant alteration of OLT performance, when compared to vehicle infusion. These results support the hypothesis that the GnRH antagonist Antide affects spatial memory via regulation of LH levels. Together these data provide strong evidence that LH acts at the hippocampus as a key modulator of hippocampal-­‐dependent spatial memory. Email: [email protected] 22 TRAVEL AWARD sponsored by ADInstruments Eric Jang Carlos Aizenman A computational model of collision detection in the optic tectum in Xenopus tadpoles E.V. JANG, A.S. KHAKHALIN, C.M. CIARLEGLIO, C.D. AIZENMAN Brown University Neural circuits in the optic tectum of Xenopus tadpoles are selectively responsive to visual stimuli that represent objects approaching the animal at a collision trajectory (looming stimuli). This stimulus selectivity is known to underlie an adaptive collision avoidance behavior in this species. While we recently showed that the balance of excitation and inhibition has a crucial role in enabling stimulus selectivity in the tectum, it is still unclear how the balance between the recurrent network activity and the newly arriving sensory flow is achieved in this structure. More generally, it is still unknown how the looming stimuli are encoded and detected by the tectal circuits, and also, despite the clear indication for the presence of strong recurrent excitation in the tectum, the exact topology of these recurrent feedback circuits remains elusive. Recently we completed a comprehensive census of tectal cell electrophysiological properties, by measuring and analyzing 30+ synaptic and intrinsic excitability parameters in each of 200+ cells from 80+ experimental animals from different developmental stages and during homeostatic plasticity. When combined with over a decade’s worth of electrophysiological analysis from our and other laboratories, we have an incredibly rich data set describing the development of tectal cell physiology during key developmental time periods. In this work we use this data set to build a high fidelity spiking network model of the tectum with the goal of generating predictions about the topology of recurrent connections within the tectum, as well as the dynamics of this system. After tuning and calibrating this model on experimental data, we compare different patterns of recurrent network connectivity, and different levels of balance between recurrent activity and sensory flow, to identify the range of parameters in which the network exhibits selectivity for looming stimuli. This allows us to make predictions about the topology of recurrent connections in the biological optic tectum that can then be tested experimentally. We also show how intrinsic excitability of individual tectal cells affects the selectivity of the network as a whole, and describe how homeostatic modulation of intrinsic properties can change selectivity thresholds in this model, thus affecting the behavior of the animal. Email: [email protected] 23 TRAVEL AWARD sponsored by the Grass Foundation Maura Schlussel Lisa Gabel The need to be me: Influence on participant specific instructions on mu-­‐based BCI performance M. SCHLUSSEL, A. BATTISON, T. FULLER, V. CORBIT, Y.-­‐C. YU, L. GABEL Lafayette College Brain-­‐computer interface (BCI) technology is a growing field, becoming an increasingly viable aid for individuals who have lost normal neural motor output. The mu rhythm, a sensorimotor rhythm that is 16 suppressed when one imagines motor activity, has been particularly utilized in BCIs because of its potential for diverse applications. While many novel implementations of mu BCIs have been developed, little work has been conducted investigating how to improve the neural signal coming from participants. Previous research from our laboratory suggest that specific instructions for imagined movement and relaxation improves overall strength of mu rhythms. Improved mu power is hypothesized to improve performance on a mu-­‐based BCI device. However it is important that sustained control over mu power, rather than maximum strength of mu rhythm is attained in order to successfully operate a BCI device. The current study aimed to improve participants’ mu BCI performance by giving them participant specific instructions for imagine motor behavior for improved control over sustained mu power. Based on previous research from our lab participants were able to successfully produce my rhythms in reponse to imagined motor or relaxing behavior in a single trial. Building upon these methods participants were provided with instructions (non-­‐specific, specific, or participant specific instructions on how to imagine motor behavior) and the ability to control the strength of the mu rhythm was analyzed. The BCI system algorithm calculated a bilateral mu power value from two electrodes positioned over the left and right sensorimotor cortices and compared to a participant’s baseline value to determine if the power increased or decreased; this difference corresponded to feedback shown on a computer screen. Preliminary evidence suggested there is a significant difference in control over mu power based on the type of instructions provided to the participant. These data may suggest that the success of an individual using a mu-­‐based BCI device may depend on the type of instructions provided. Decreasing training time, increasing BCI literacy, and enhancing control over mu-­‐based BCI devices will make this type of BCI device more accessible to individuals with impaired motor behavior. Email: [email protected] 24 Cassie Lincoln Michele Lemons Characterizing the role of integrins in axon pathfinding in i C. elegans C.A LINCOLN, D.O OLIVER, M.M FRANCIS, M.L LEMONS Assumption College The ability of neurons to properly extend axons during development through a complex extracellular milieu and ultimately reach their appropriate targets is remarkable. The molecular mechanisms that drive this impressive navigational feat are not yet fully understood. Previous studies suggest that a family of transmembrane heterodimeric proteins, called integrins, play an important role in neuronal motility. We chose to more fully characterize the effects of integrins on axon patterning in the genetically-­‐powerful model organism, Caenorhabditis elegans. This model organism enables us to study axon patterning in vivo. C. elegans is an advantageous model to use for these studies due to its limited number of integrin subunits (two alpha subunits and one beta subunit) compared to vertebrates (18 alpha subunits and 8 beta subunits). We initially examined the effects of the alpha subunit ina-­‐1 upon axon guidance in a hypomorphic strain, ina-­‐1(gm144). This allele is thought to contain a mutation that is important for integrin activation. Integrin activation refers to a conformational change that induces a high ligand affinity state. Using a genetic approach, we found that decreased integrin function in ina-­‐1(gm144) animals produce: 1) strong defects in the patterning of commissural axons from GABAergic neurons, 2) less robust effects on commissural axons from cholinergic neurons, and 3) no obvious effects on longitudinal axons projecting from the tail to the head (e.g. from the interneuron DVA). Interestingly, errors in GABAergic and cholinergic axon patterning were most pronounced at distinct times during development. This evidence suggests that the role of ina-­‐1 is not equal in all neurons, nor is the contribution of integrins equal over time. Confocal analysis reveals that INA-­‐1 is expressed in GABAergic neurons. Future studies will determine if ina-­‐1acts cell autonomously by driving wildtype ina-­‐1 expression in GABAergic neurons in ina-­‐1(gm144) animals. We will also expand our studies to include an examination of the possible influence of the two remaining integrin subunits (pat-­‐2 and pat-­‐3) on axon guidance in C. elegans. These studies will advance our understanding of the role of integrins in axon pathfinding of GABAergic and cholinergic neurons. This work will help unravel the molecular mechanisms of axonal navigation during development and the findings from this work could be used to 17 enhance axonal regeneration of injured adult neurons. The latter could be beneficial in designing improved treatment for spinal cord injuries or stroke in vertebrates. This work was funded by a AREA R15 1R15NS070172-­‐01A1 awarded to M.L. Lemons. Email: [email protected] 25.1 Ashley Ealey Emily Hardy Arl13b deletions effect on postnatal cerebellar development A. EALEY, S. BAY, T. CASPARY Agnes Scott College Our lab studies Arl13b, a small GTPase that is found in the primary cilium of cells and regulates Sonic hedgehog (Shh) signaling. Shh signaling is dependent on primary cilia. The primary cilia and appropriate Shh signaling are important for development of the cerebellum because they direct proliferation of granule neuron precursor cells. Mutations in Arl13b are linked to human disorders such as Joubert Syndrome as well as medulloblastomas, tumors created by overactive Shh signaling during cerebellar development. At prenatal stages of development, Arl13b is essential for normal cerebellar morphology. Deletion of Arl13b early in development alters Shh signaling compromising cerebellar morphogenesis, resulting in a small or absent cerebellum. We hypothesized that Arl13b could also play a role in postnatal cerebellar development. We deleted Arl13b in four day old mice in the granule neuron precursor cells. We compared brain tissue from control and experimental animals and looked for expression of Arl13b in the external granular layer and gross morphology. Our findings allow us to conclude that deletion of Arl13b at P4 does not appear to have a gross impact on postnatal cerebellar development. 25.2 Alanah Grisham Emily Hardy Mutagenesis of the G-­‐Quadruplex in the BDNF 3' UTR reduces BDNF reporter expression A. GRISHAM, M. ALLEN, Y. FENG Spelman College Brain Derived Neurotrophic Factor (BDNF) is a secretory protein in the brain that plays multiple roles in normal brain function as well as in multiple neurological diseases. Alternative polyadenylation of the BDNF transcripts results in two types of BDNF mRNA: containing either a short or a long 3’ untranslated region (UTR). Sequence analysis identified a guanine rich element, which is located close to the proximal polyadenylation site in the BDNF transcript that can form a G-­‐Quadruplex structure. It is known from examples in cancer research, that G-­‐Quadruplex structures in DNA can affect transcription. We hypothesize that the G-­‐Quadruplex in the BDNF 3’UTR regulates BDNF expression. In this summer research program, we explored the function of the G-­‐Quadruplex using a luciferase reporter that contained the BDNF long 3’ UTR. We discovered the first evidence that the G-­‐Quadruplex plays an important role in BDNF expression. 26 Sharena Rice Kenneth Owen Long Localization of the ABCA4 flippase in developing photoreceptors S.P. RICE, K.O. LONG California Lutheran University 18 Introduction: Flippases are proteins that transport lipids across cell membranes and serve as key regulators of cell polarity. There are a number of different flippases in the retina. Flippases can be degraded with time by over-­‐oxidation, and mutations can cause flippases to be absent. A defect in the photoreceptor-­‐specific flippase ABCA4 is linked to Stargardt macular degeneration in humans. In the adult fish retina, there are developing photoreceptor cells at the periphery of the eye (close to the iris) along with fully differentiated photoreceptors throughout the rest of the retina. Fish were the model organisms because 1) their eye anatomy is similar to that of humans and 2) they have indeterminate growth so that I can compare an individual’s newer photoreceptors to its older ones. Using goldfish retinas allow the visualization of the developmental appearance of ABCA4 and related proteins in photoreceptors. Purpose: The purpose of this study was to determine the effectiveness of different commercial antibodies in labeling flippases (ABCA4 and ATP8a2) in the goldfish retina. These antibodies were not known to work in fish retina before. The ultimate goal is to analyze the developmental appearance of flippases in photoreceptors and whether they appear before or after opsin, the light-­‐
capturing protein of photoreceptors. Materials and methods: Commercial goldfish were used. SDS-­‐PAGE and Western blotting were used to determine the molecular weights of the proteins and to tell which antibodies bound proteins of the goldfish retina. The antibodies tested included Abcam's monoclonal anti-­‐ABCA4, GeneTex’s polyclonal anti-­‐ABCA4, Santa Cruz Biotechnology's anti-­‐ATP8a2 (a related flippase) and anti-­‐opsin. Light and dark-­‐adapted retinas were also fixed and processed for immunocytochemistry. Results: This study demonstrated that GeneTex’s polyclonal anti-­‐ABCA4 is effective in labeling ABCA4 in goldfish retinas. Santa Cruz Biotechnology’s monoclonal anti-­‐ABCA4 does not work as well, as it requires a great concentration of these antibodies to work. Further studies will analyze the localization of the ABCA4 in the developing retinas. Preliminary immunocytochemical results demonstrated ATP8a2 labeling of developing photoreceptors. The anti-­‐opsin also worked well in labeling. Conclusion: Since the polyclonal anti-­‐ABCA4 from GeneTex and the anti-­‐ATP8a2 work in the goldfish retina, they can be used in further studies to find more about these flippases and their developmental appearance in the retina. Email: [email protected] 27.1 Kayla Roberge Matthew L Beckman Studies of the role of hedgehog signaling in daphnid eye development K. ROBERGE, M. GRAFELMAN, C. GIDDINGS, M.L. BECKMAN Augsburg College, Minneapolis, Minnesota (MN) Daphnia magna are freshwater invertebrates that are used to study effects of toxins on development and reproduction. An important part of their development involves the apparent fusion of two separate eyespots into a single, central eye that occurs in early embryonic development. . The mechanics behind this fusion are not entirely understood nor are the genes involved, but preliminary data I suggests that the hedgehog signaling pathway is involved. In humans there are 3 hedgehog family genes including sonic hedgehog. Mutations in sonic hedgehog is associated with a variety of midline defects. A known inhibitor of the hedgehog pathway, U18666A, has been used in rats and found to be a cholesterol synthesis and hedgehog signaling inhibitor. The effects of U18666 on the development of Daphnia magna embryos was assessed. Data collected in this study of embryos growing in culture demonstrated that there were developmental delays as well as a delay in eye fusion when early stage embryos were treated with 100 µM drug. There were no significant results obtained from the in vivo treatment of pregnant females. These results signify a need to further evaluate the effect of Hedgehog signaling pathway modulators on Daphnia magna embryonic development. http://beckmanlab.org Email: [email protected] 19 27.2 Bobby McDonnell Matthew L Beckman The role of dopamine in the appendage movements associated with swimming and feeding in daphnids B. MCDONNELL, R. SNEED, M.L. BECKMAN Augsburg College, Minneapolis, Minnesota (MN) Daphnia magna, commonly referred to as the water flea, and D. pulex have been utilized extensively in studying toxicology. Both have also been studied with respect to their locomotor activity under unconstrained conditions, but limited research has been done on D. magna with a focus on the effect dopaminergic drugs have on force production. Animals were filmed with two orthogonal cameras focused on the displacement of a plastic fiber on which the specimen was mounted. The animal was filmed over a period of three hours divided into three segments, in which the animal’s initial force production abilities were observed, then a dopaminergic drug (A-­‐68930) was added to assess the drug’s effect on the animal, and lastly a washout period ensued to observe the returned force production potential as the drug is washed out of the animal. Force production was observed to be reduced with drug exposure. The maximum force produced after 1 hour in 80μM A-­‐68930 was statistically significantly smaller than in the pre-­‐drug state (baseline). A trend toward drug washout was observed but was not statistically significant. Future studies will focus on increasing the sample size and carrying out a thorough dose-­‐response for A68930 using separate groups of control and drug-­‐treated animals. These experiments demonstrate a role for dopamine signaling in modulating Daphnia magna swimming. http://beckmanlab.org Email: [email protected] 28 Joshua Leonard Nadja Spitzer Fructose impairs neuronal differentiation of adult neural stem cells in vitro J.A. LEONARD, A.L. RAMIREZ GARCIA, N. SPITZER Marshall University In recent decades the consumption of fructose, as high fructose corn syrup in processed foods, has increased dramatically and has now been recognized as a significant health concern. In hepatocytes and adipocytes, high levels of fructose generate uric acid, leading to generation of reactive oxygen species (ROS) that cause cellular damage. Fructose penetrates the blood brain barrier through the Glut5 transporter, but its effects on the mechanisms operating in neurons is not well understood. In animals exposed to a high-­‐fructose diet, cognitive deficits similar to those associated with aging are observed. Reduced neurogenesis is thought to contribute to the cognitive decline in aging, indeed, a high-­‐fructose diet also reduces adult neurogenesis, although the cellular mechanisms by which this occurs is unclear. In mammals, defined populations of adult neural stem cells (NSCs) are located in the hippocampus and the subventricular zone (SVZ). Here, they continue to proliferate, giving rise to progenitor cells that migrate, differentiate and integrate into existing circuitry where they are involved in plasticity with roles in memory formation, learning, behavioral responses, and reward systems. In addition, NSCs are thought to migrate to damaged brain tissues and contribute to repair. In vivo and in vitro, NSCs can differentiate into oligodendrocyte, astrocyte, or neuronal phenotypes in response to appropriate chemical signals. Adult NSCs from the SVZ can be maintained as progenitors in culture and plated in conditions that encourage differentiation, allowing investigation of the cellular mechanisms underlying this process at the level of individual cells. This accessible model system has been used to identify many of the exogenous signals that drive differentiation, and the intracellular mechanisms involved. We tested the effects of fructose exposure on the mechanisms underlying differentiation in NSCs cultured from the SVZ of young adult rats. Using immunocytochemistry, we found that the proportion of cells expressing β-­‐tubulin III, an early neuron marker, was reduced in cells cultured in media containing fructose. This indicates that fewer NSCs differentiate towards a neuronal lineage after fructose exposure. Time lapse microscopy revealed that the 20 neurites extended by NSCs cultured in the presence of fructose are less complex, with fewer branch points than those differentiating in control conditions. These results suggest that fructose inhibits the maturation of NSCs towards a neuronal fate. These cellular effects on differentiating NSCs could contribute to the cognitive decline observed in animals subjected to a high-­‐fructose diet. Email: [email protected] 29 Mir Shanaz Hossain Frances Northington Imaging the spatiotemporal progression of white matter injury after neonatal hypoxic-­‐
ischemia M.S. HOSSAIN, J. BURNSED, R. CHAVEZ-­‐VALDEZ, J. ZHANG, F. NORTHINGTON Johns Hopkins School of Medicine Background: Hypoxic-­‐ischemic encephalopathy (HIE) is a leading cause of death in children under age 5. Hypothermia therapy (HT) is the only treatment specific for HIE, however, the neuroprotection provided is incomplete. MRI is an important clinical tool in injury assessment and outcome prediction following HIE. Conventional MRI is used to characterize injury; now, white matter (WM) injury can be detected with diffusion tensor imaging (DTI). Using a newly established model of HT in the neonatal mouse model of HIE, the extent of WM injury following HI and HT was determined with MRI. Methods: HI was induced in postnatal day (P)10 mice using a modified Vannucci Model (unilateral carotid ligation & 45 minutes of hypoxia at FiO2= 0.08). Following HI, the mice recovered at 31°C (HT) or 36°C (Normothermia Therapy, NT) for 4 hours. T2-­‐weighted and DTI MRIs were obtained from injured and control mice at P30. Amira software was used to quantify the volume of major gray matter structures. The gray matter volumes were compared to the volume and mean fractional anisotropy (FA) values of related white matter tracts, obtained using ROIEditor. Results: NT mice had reduced ipsilateral hippocampal volumes (5.845 vs. 12.07, p < 0.001) and lower ipsilateral fimbria FA values (0.4402 vs. 0.5868, p< 0.001). NT mice had smaller ipsilateral cerebral cortices (52.09 vs. 64.38, p < 0.01) and lower corpus callosum FA values (0.534 vs. 0.5952, p < 0.01). The fimbria volume/FA values and the hippocampal volumes were significantly correlated (R2 = 0.54488 and 0.74047, respectively; p < 0.01 for both) in the NT mice, however, no correlation was found between the corpus callosum volume/FA values and cortical volumes. Similarly, HT mice had reduced ipsilateral hippocampal volumes (7.128 vs. 12.07, p < 0.001) and lower ipsilateral fimbria FA values (0.4677 vs. 0.5868, p< 0.001). Ipsilateral cortex volume (54.63 vs. 64.38, p < 0.05) and corpus callosum FA values (0.5507 vs. 0.5952, p < 0.05) were lower in the HT mice. The fimbria volume/FA values and the hippocampal volumes were correlated (R2 = 0.4482 and 0.77795, respectively; p < 0.01 for both), and a correlation between the corpus callosum and cortical volumes was found (R2 = 0.23979) in the HT mice. Stratifying for gender revealed that significance in hippocampal volumes and fimbria FA values are attributable to male mice, with no differences in female mice treated with NT and HT. Discussion: The findings from this study show that important differences in WM volume and structure are detectable following neonatal HI, especially in male mice. The results from this study can be used to improve the quantitative evaluation of HI injury and therapeutic efficacy. Research Support: Nu Rho Psi Undergraduate Research Grant, Johns Hopkins University Second Decade Society Internship Grant Email: [email protected] 30 Elizabeth Hughes Skyla M. Herod Differential effects of maternal and fetal hyposerotonemia on placental physiology, biogenic amine function and the co-­‐expression of 5-­‐HT/NetrinG1 in the developing fetal brain. E. HUGHES, J. VALASQUEZ, C. CRONKITE, K. FISH, E. EMERY, A. BONNIN, S.M. HEROD Azusa Pacific University 21 Serotonin (5-­‐HT) is a monoamine neurotransmitter that is fundamental to brain development, and low circulating levels of 5-­‐HT are associated with clinical psychopathologies including anxiety and depression. Interestingly, the placenta, and not the maternal blood or fetal brain itself, is responsible for providing 5-­‐
HT to the fetus early on in development. It remains unclear whether hyposerotonemia in mothers with such pathologies might affect fetal brain development, placental levels of 5-­‐HT, and other biogenic amine signaling. B6.129(Cg)-­‐Slc6a4tm1Kpl/J serotonin transporter knockout (SERT-­‐KO) mice were bred heterozygous (HET) by HET to generate wildtype (WT), HET, and knockout (KO) experimental genotypes. Females of each genotype were bred with HET males, and fetal and placental tissue was harvested at four different timepoints during embryonic development (E12.5, E14.5, E16.5, E18.5) as part of ongoing studies. High performance liquid chromatography (HPLC) has been performed for analysis of concentrations of 5-­‐HT and 5-­‐HIAA, the main metabolite of 5-­‐HT, present in E14.5 and E18.5 fetal forebrains, hindbrains, and placentas across all maternal and fetal genotypic combinations. Significant differences appear in tissue samples from the same maternal genotype regardless of fetal genotype, suggesting that the mother may be the more influential factor in the determination of fetal serotonergic system functioning. Fetal brains and placentas were also collected for immunohistochemical analysis, and a costain was developed for 5-­‐HT and Netrin-­‐G1 (NetG1), an axon guidance cue instrumental in the directing of thalamocortical axons (TCAs) during fetal brain development. Upon fluorescent imaging to determine the cellular coexpression of these two essential molecules, initial analysis of WT and KO fetal brains at E14.5 from the same HET mother showed no differences in the 5-­‐HT neuronal expression, consistent with the HPLC data. However, the expression of the NetG1 in the KO was attenuated and less fasciculated in the midbrain region as compared to WT. These results suggest that while 5-­‐HT may not be affected by fetal genotype, production and/or expression of NetG1 may indeed be affected by fetal hyposerotonemia, possibly under the influence of a different mechanism. Sources of Support: Beta Beta Beta Foundation Grant, Azusa Pacific University Faculty Research Grant, University of Southern California Faculty Research Startup Funds Email: [email protected] 31 Kyle Fish Skyla M. Herod Interaction of SERT deletion, early life stress and maternal presence as indicators of individual differences in the corticosterone, biogenic amine signaling, and adulthood behavioral response to stress in B6.129(Cg)-­‐Slc6a4tm1Kpl/J (SERT-­‐KO) mice. K. FISH, E. EMERY, E. HUGHES, J. VELASQUEZ, K. LEWANDOWSKI, A. BARAJAS, A. BONNIN, S.M. HEROD Azusa Pacific University Circulating serotonin levels and exposure to stress during critical periods of development may play a role in determining an individual’s sensitivity to stress, and shaping the development of stress related disorders. The current study examined the interactive effects of early life stress exposure and chronic serotonin deficiency on stress-­‐sensitivity in mice. Potential protective effects of maternal presence during early life stress were also investigated. WT, HET, and KO genotypes of the B6.129(Cg)-­‐Slc6a4tm1Kpl/J (SERT-­‐KO) mouse strain were used as a genetic model of serotonin depletion. Mice were subject to a stress paradigm consisting of moderate restraint stress exposure during early adolescence, with or without maternal presence, followed by a stress test of similar nature upon maturity to adulthood. Initial analysis has shown a genotype by stress condition interaction in both the adolescent and adulthood corticosterone (CORT) response. In WT mice, but not HET or KO, stress exposure in adolescence led to an elevated CORT response compared to controls (F=11.94, p=0.00). Interestingly, mice exposed to stress during adolescence with maternal or novel female presence showed a heightened CORT response compared to animals stressed alone. In adulthood, a genotypic difference was observed among the animals originally exposed to stress with maternal presence, with WT animals showing the greatest CORT response compared to other genotypes within this treatment group (F=3.57, p=0.05), indicating that the effects of serotonin deficiency may depend on the specific environmental setting. In a light/dark box test, 22 KO mice spent less time in the light (F=2.97, p=0.05) and exhibited fewer line crossings (F=17.61, p=0.00) than WT or HET mice, indicating diminished interest in exploratory activity and increased anxiety-­‐like behavior, across all stress conditions. In additional behavior tests, KO mice across all stress conditions made fewer social approaches (F=9.53, p=0.00) and exhibited increased depressive-­‐like behavior (F=2.96, p=0.05). Group differences were also observed in multiple measures of monoamine (DA, 5-­‐HT, NE, E, their precursors and metabolites) concentration in various brain regions. The high level of individual variability in the CORT response in each of the 12 treatment groups across development, coupled with the lack of interactive effects in behavioral measures, suggests that these factors alone may not be the most powerful components that shape stress-­‐sensitivity in adulthood. Email: [email protected] 32 Joseph Wetherell Mark Jareb The role of α6β4 integrin receptor in laminin-­‐mediated axon outgrowth J. WETHERELL, A. BUONACCORSI, N. DOPPLER, M.I. JAREB Sacred Heart University Previous data suggest that a laminin receptor is localized in the axons of chick forebrain neurons mediating the axon-­‐growth promoting properties of laminin. Many heterodimers from the integrin family of proteins have previously been identified as laminin receptors, including the α6β4 heterodimer. We tested the role of β4, β1, and α6 integrins in laminin-­‐induced increases in axonal growth using function blocking antibodies. Axons of neurons grown on laminin treated with β4 integrin function blocking antibodies or α6 integrin function blocking antibodies were significantly shorter compared to untreated cultures or cultures treated with a β1 integrin function-­‐blocking antibody. These data are consistent with the hypothesis that the α6β4 heterodimer acts as the axonal laminin receptor in embryonic chick forebrain neurons and is important in axonal development and growth. To directly test whether α6 or β4 integrin was localized specifically to axons, we transfected cultured forebrain neurons from embryonic chick with DNA constructs encoding both α6 and β4 integrin genes. Preliminary results show α6 and β4 integrin expression in axons. Email: [email protected] 33 Spencer Duncan Jacob Cain SGEF-­‐mediated Rho GTPase signaling controls neural progenitor proliferation and migration in the developing cortex. S. DUNCAN, J. CAIN, S. KOH, D. TIMMS, J. WEIMER Sanford Research During the development of the cerebral cortex, neural progenitors proliferate and then migrate away from the ventricular zone (VZ) to take up residence in the cortical plate (CP). These immature cells take one of two paths upon exiting the VZ: 1) migrating directly to the CP, or 2) migrating to the subventricular zone (SVZ), becoming intermediate progenitors (IPs), dividing several more times, and then continuing to the CP. Although disruptions in these events are known to contribute to neurodevelopmental disorders, the factors regulating progenitor retention in the SVZ are relatively unknown. Small Rho GTPases, including cdc42 and RhoG, are known to regulate progenitor proliferation and fate determination within the developing cortex. However, the regulators of these Rho GTPases are not well understood. GEFs and GAPs are proteins that regulate Rho GTPase activity, and because GEFs act as activators for Rho GTPases, we hypothesize that GEFs are critical to Rho GTPase-­‐mediated regulation of progenitor proliferation and fate determination in the developing cerebral cortex. To explore the role of one GEF identified as a regulator of cdc42 and RhoG, SGEF, in the process of IP migration and release, IHC was performed on brains from an 23 SGEF -­‐/-­‐ mouse model across several time points to determine differences from wild type brains in the areas of proliferation, migration, and layer thickness. In addition, to explore how SGEF is genetically regulated, a promoter region was found and confirmed and potentially-­‐binding transcription factors were identified. In the future, these transcription factors will be confirmed through site-­‐directed mutagenesis of the DNA binding sites to further confirm the signaling pathway identified in these experiments. Email: [email protected] 34.1 Benjamin Foster Katie Wiens Neural innervation of zebrafish heart. B.P.FOSTER, E.N.COSTANZI Christopher Newport University Zebrafish have the ability to to regenerate various parts of their bodies including neural and cardiac tissue. This study seeks to understand the mechanism of zebrafish cardiac regeneration by determining the neural pathways that innervate the zebrafish heart. Using Tyrosine Hydroxylase (TH), Acetylcholinesterase (Ach), Substance P (P), neural Nitric oxide Synthase (nNOS), and Galanin stains, this study has found presence of various neurotransmitters in the adult zebrafish heart, thus indicating the presence of neural input within zebrafish hearts. The presence of the aforementioned neurotransmitters were verified by fluorescent microscopy. The presence of Tyrosine Hydroxylase indicates the presence of dopaminergic terminals, acetylcholinesterase of acetylcholine terminals, Substance P of Substance P receptors, nNOS of NO activity, and galanin of synaptic regulation. Email: [email protected] 34.2 Andrew Moyer Katie Wiens Zebrafish model to test the effect of copper on amyloid beta precursor protein. A. MOYER, K. DOAN, B. SOMERA, D. KNIGHT, K. WIENS Christopher Newport University The buildup of amyloid-­‐beta (Aβ) plaques is a hallmark of Alzheimer’s disease. Recent studies have investigated compounds that break down Aβ plaques and reverse the cognitive decline associated with Alzheimer’s disease. However, we have yet to discover a treatment that prevents the progression of this devastating disease. Zebrafish are an excellent model system for investigating treatments for Alzheimer’s disease; zebrafish are inexpensive, produce hundreds of embryos each day, and have homologous genes related to human Alzheimer’s disease. The research presented aims to develop a copper-­‐induced model of alzheimer's disease in the zebrafish. This model will allow us to study the effect of multiple compounds on Aβ plaque formation in an environmentally-­‐induced zebrafish model of Alzheimer’s disease. Email: [email protected] 35 Maribel Santos Adam Hall Isoflurane impacts the actin cytoskeletal by upregulating cofilin phosphorylation in neonatal murine cortical neurons M.O. SANTOS, J. BARGER, B. NHUNDU, A. HALL Smith College 24 For the past decade, general anesthetics have been under more intensive scrutiny as evidence for their potential detrimental effects in the brains of both the elderly and children have come to light. Recent work in animal models has linked commonly used anesthetics to inducing neurotoxicity and subsequent neuronal cell death. Indeed, we identified that isoflurane at clinical concentrations induces morphological changes in murine neurons in culture that could impact activity-­‐dependent survival of developing neurons. The aim of current study is to elucidate the cellular mechanism responsible for these morphological and potentially neurodegenerative effects of isoflurane. Using mice exposed to clinical concentrations of isoflurane (1.5% v/v for 0.5 hr), we hypothesized that the anesthetic would induce morphological alterations of the cytoskeleton (and subsequent neurodegeneration) through an up-­‐
regulation of RhoA (GTPase) activity that increased LIM Kinase-­‐1 activity with subsequent phosphorylation of cofilin, a key modulator of actin dynamics. To date, using immunoblotting techniques, we have identified a significant increase in phosphorylated cofilin and in phosphorylated LIM Kinase in treated murine brain samples relative to non-­‐treated. To confirm that the impact of isoflurane on cell morphology is via changes in RhoA activity, in future experiments we plan to inhibit the activity of LIM Kinase in cultured neurons and observe if anesthetic-­‐induced changes in cell morphology are arrested. This study will help expand our knowledge of anesthetic toxicity by elucidating a mechanism by which isoflurane induces cytoskeletal rearrangements that may lead to neurodegeneration, with the ultimate goal of minimizing damage to at-­‐risk populations during surgery. Email: [email protected] 36 Heather Huffman Joseph L. Cheatwood Rhox8 expression in rodent brains H.G. HUFFMAN, D. E. GRISLEY, J. A. MACLEAN, J. L. CHEATWOOD Southern Illinois University Homeobox genes encode transcription factors that govern many processes during development. This 60-­‐
amino acid DNA-­‐binding motif associates with promoters by either activating or suppressing the transcription of downstream target genes. Recently, the Rhox, -­‐(X-­‐linked reproductive Homeobox)-­‐, genes were discovered. The Rhox gene set is expressed during embryonic development in the testis, but a select few of these genes continue to show high expression after birth which makes them candidates for controlling postnatal and adult developmental events. At their peak expression in the testis (postnatal day 12), all Sertoli cells express Rhox5, Rhox8, and Sox9, but Rhox5 expression become more restrictive after postnatal day 30. RHOX8 protein is abundant in the testis, epididymis, ovary, and it is weakly detected in the placenta. Of all 33 mouse Rhox genes, Rhox8 is the only one to show expression in somatic cells in the embryonic testis. All others are expressed solely in germ cells. It is a common phenomenon that testis expressed genes exhibit brain-­‐specific splicing or transcripts from alternative promoters. Given Rhox8’s potentially unique transcriptional control in the testes, we wanted to determine if Rhox8 was similarly uniquely expressed in the brain. Thus, we examined whether the somatic transcription factors Rhox8, Sox9, and Rhox5 mRNA could be detected in rat mouse brain tissue. Rats were euthanized, and brains were removed and flash-­‐frozen. To study the basic expression of the Rhox8, Sox9, and Rhox5 gene in brain tissue, we extracted RNA from the rat cerebellums, and then converted the RNA to cDNA in order to run qPCR, quantitative real-­‐time polymerase chain reaction, on. Preliminarily Rhox8, Rhox5, and Sox9 were all found to be highly expressed in positive control adult mouse testis tissue. Of these only Rhox8 had moderate expression in mouse cortex. While RHOX8 stained positive in mouse neurons. As expected we found Rhox8 to have moderate expression in adult rat cerebellum, Sox9 had minimal expression, but Rhox5 had no detectable expression. These low levels may be due to the use of whole cerebellum in the rat, which contains many cell types which do not appear to express Rhox8 in addition to neurons, which stain positive for RHOX8 via fluorescent immunohistochemistry in mice or mouse cortex. Email: [email protected] 25 37.1 Biagio Niro Jacqueline Morris Inhibition of PAD 2 and its implications in neuronal development B. NIRO, G. RANCHOFF Baldwin Wallace University Peptidylarginine deiminase 2 (PAD 2) converts arginine residues in myelin basic protein (MBP) into citrulline. An increase in citrullinated MBP is present in multiple sclerosis patient nervous tissue and may have a role in the disease. In addition to an increase in disease, PAD 2 is a protein that is present early in development but the importance of this enzyme during development is unknown. Since PAD 2 is present early in development and then re-­‐emerges during disease states, the role of PAD 2 during this critical stage might provide insight into this increased expression. In order to determine the role of PAD 2 in early development, we inhibited PAD 2 in zebrafish embryos (2-­‐24 hpf) with 2-­‐chloroacetamidine (0.5 mM, 1.0 mM, 1.5 mM). The embryos (24 hpf) were fixed and stained for acetylated-­‐tubulin to mark the nervous system. It was determined that zebrafish treated with 2-­‐chloroacetamidine had fewer Rohon Beard cells, early sensory neurons, compared to controls. Rohon Beard (RB) cells are a transient population and generally undergo apoptosis at 5 days post fertilization (dpf). After RB cell death, dorsal root ganglia develop, therefore, the PAD 2 enzyme or citrullination may be involved in neuronal differentiation of sensory neurons. Email: [email protected] 37.2 Ashley Mahajan Jacqueline Morris The effects of ketamine on learning and spatial memory in juvenile rats A.P. MAHAJAN, C. SMITH, J.M. BROWN, B. NIRO, J. SMITH, J. MORRIS Baldwin Wallace University During early childhood, the brain is susceptible to environmental changes which may result in cell death. Ketamine is an anesthetic agent used in children (< 4 years), which has shown to cause increased apoptotic cell death in the hippocampus in neonatal rats (P7). Previous studies indicate that the Ketamine-­‐
induced apoptosis occurs with binding of Ketamine to N-­‐methyl-­‐D-­‐aspartate (NMDA) receptors which disrupts the neural uptake of calcium. The aim of this study was to determine if ketamine adversely effects spatial memory or reasoning in neonatal rats. A prospective trial was conducted utilizing 25 Sprague-­‐
Dawley rats divided into a control and experimental group. On P7 animals were administered four subcutaneous doses of ketamine (20 mg/kg) or an equivalent amount of sterile saline. At P25, both groups were tested using the Morris Water Maze to assess their spatial learning and memory. During training days, each rat had four trials to locate the escape platform before the final probe trial when the platform was removed from the pool. The average time spent in the platform-­‐containing quadrant once the platform was removed was 30.74 seconds for the control and 28.30 seconds for the experimental rats (P= 0.792). Data suggests that learning for both groups was variable. Early exposure to ketamine did not alter ability of the animals to learn using spatial memory in the Morris Water Maze. Therefore a loss of neurons early on may be compensated by new neurogenesis. Additionally other forms of learning might be compromised and will be studied with other learning tasks. Email: [email protected] 26 38 Melissa Smallwood Tyisha Williams Synaptic troubleshooting: Making the connection between autism and labor & delivery drugs M. SMALLWOOD, E. BAKER, A. SAREEN, R. HANNUSCH, T. WILLIAMS Trinity University Autism Spectrum Disorder (ASD) is a pervasive neurodevelopmental disorder characterized by social deficits, difficulties in communication, and repetitive behavior. The recent increase in the rate of autism diagnosis in the United States – 1 in 68 children as of 2014 – has sparked interest within the research community to investigate potential causes and risk factors involved in the development of autism. Genetic and environmental factors – and interactions between them – have been implicated in the development of the autistic phenotype. It is believed that environmental stressors or toxins could modify the expression of important genes and proteins in brain development. Previous research assessing environmental factors related to autism indicates that a relationship may exist between autism and the labor-­‐inducing drug, Pitocin, during childbirth. Therefore, the aim of this study was to determine if epidurals and/or labor-­‐
inducing drugs are potential risk factors for autism. Results from our population study show that children exposed to labor and delivery drugs during childbirth are 2.64 times more likely to develop an autistic phenotype (X2 = 3.88, p = 0.048). The data also show a relationship between duration of exposure and the chance of developing an autistic phenotype (X2 = 4.88, p = 0.027, OR = 2.64). In order to assess the possible molecular effects of labor and delivery drugs, the expression patterns of six autism candidate genes -­‐ UBE3A, NLGN3, NLGN4, SHANK, CNTNAP2, and GABRB3, were assessed at various mouse developmental stages (Postnatal day 0, 7, 14, and 21). Indeed, altered gene expression was observed for all six target genes, with the most drastic changes occurring at postnatal day 0 and 7. The data suggest that exposure to labor and delivery drugs during childbirth could potentially interfere with normal expression of critical neurodevelopmental genes, such as those involved in synapse formation. Future behavioral studies are required to evaluate the long-­‐term neurodevelopmental consequences of these drugs that could increase the risk for the development of autism. 39 Pompeyo Quesada Veronica G. Martinez Acosta Gene transcript analysis of potential Lumbriculid regenerative proteins through the development of a QPCR assay P.R. QUESADA, R.A. MIRANDA, J. ARJONA-­‐SOBERON, V.G. MARTINEZ-­‐ACOSTA University of the Incarnate Word The aquatic oligochaete Lumbriculus variegatus is an exceptional regenerating model system. To further understand the regenerative mechanisms utilized by Lumbriculus, we are investigating genes and proteins that are known to regulate regeneration in other species. Our lab is particularly interested in regeneration within the nervous system. Previous data demonstrates an up regulation of the protein β-­‐
catenin within the worm’s central nervous system during regeneration. β-­‐catenin is known to function as a transcriptional regulator in the canonical Wnt signaling pathway and also functions as an integral part of cadherin-­‐mediated cell adhesion at the plasma membrane. Thus, we would like to determine the role β-­‐
catenin may be playing during regeneration in Lumbriculus variegatus. In order to further investigate the role of β-­‐catenin and other genes during Lumbriculid regeneration, we are developing a QPCR assay to evaluate changes in transcript levels within worm tissue. Conserved regions of gene sequences encoding putative regenerative proteins in other closely related species were used to design degenerative PCR primers, which were then used to amplify Lumbriculid genes. Gel electrophoresis analysis indicate that we have PCR products of the predicted base pair size for reference genes including alpha tubulin, β-­‐actin and ribosomal protein L8. Primer sets for the genes encoding β-­‐catenin and other important regenerative proteins (eg. ZicA, FoxD, and Wnt proteins) will be developed and investigated. Collectively these findings will help us better understand the regenerative mechanisms in Lumbriculus and potentially in higher 27 order phyla. Grant Support: This work is supported in part by DoD Grant W911NF-­‐13-­‐1-­‐0164 (V.G.Acosta) and the University of the Incarnate Word Department of Biology (Dr. B.McCormick). Email: [email protected] 40.1 Bradley Gehring Sonsoles de Lacalle Neuropeptide distribution in the human parabrachial nucleus. B. GEHRING, S. DE LACALLE Ohio University The Parabrachial Nucleus (PB) is a pontine structure composed of several cell groups located around the superior cerebellar peduncle, with a crucial role in autonomic control. In the human brain, the PB is divided into a medial (MPB) and lateral (LPB) nuclei. The human PB is cell poor and does not seem to contain the many subnuclei described in the rat. Nonetheless, there are a number of chemically distinct subdivisions that as well as contributing to establish homologies, may be of value in pathological investigations. Following our earlier work using CGRP as a marker for ascending visceral pathways in the human brain (de Lacalle & Saper, 2000), here we describe the distribution of galanin (GAL-­‐), substance P (SP-­‐) and neurotensin (NT-­‐) immunoreactive elements. Our observations were made on horizontal 50μm thick sections through the brainstem from 3 neurologically normal individuals, obtained at routine autopsy. Tissue was processed for immunocytochemistry using commercially available antibodies. We found several areas of dense peptide immunoreactivity in fibers, as well as scattered stained cell bodies. The distribution of peptide-­‐stained fibers was strikingly conserved compared with that described in the rat. GAL-­‐ ir fibers were present in both subdivisions, but more abundant in the LPB than the MPB, with their distribution somewhat overlapping that of CGRP-­‐ir fibers. By contrast, NT-­‐ir fibers were more prominent in the MPB than in the LPB. SP-­‐ir fibers were found in both MPB and LPB, with a dense cluster of fibers in the rostral region, and scattered SP-­‐ir neuronal profiles throughout the area between the locus coeruleus and the MPB. Compared with the literature on the functional anatomy of the PB and its afferent and efferent projections, our results suggest that these neuropeptides found in distinct areas within the human PB may also provide chemical coding for the relay of specific visceral information. Email: [email protected] 40.2 Nicholas Lozier Sonsoles de Lacalle Interaction between myostatin and the GH/IGF-­‐1 axis in muscle N.R. LOZIER, E.O. LIST, D.E. BERRYMAN, J.J. KOPCHICK, S. DE LACALLE Ohio University Myostatin (mstn) has been identified as a negative regulator of muscle growth, leading to research into its potential as a therapeutic agent in individuals suffering from muscle wasting disorders, as occur in old age, cancer and AIDS, for example. Mstn acts locally as an autocrine/paracrine agent to inhibit muscle hypertrophy and hyperplasia, and it is widely believed that this pathway can be silenced by the endocrine action of IGF-­‐1 on skeletal muscle. Our present work seeks to understand the mechanisms by which mstn and the GH/IGF-­‐1 axis interact in muscle, using two animal models, the growth hormone receptor knockout (GHR-­‐/-­‐) and the bovine growth hormone transgenic (bGH). These mice have contrasting phenotypes. IGF-­‐1 action is significantly reduced in the GHR-­‐/-­‐ because of the lack of GH signaling, and results in obese and dwarf mice that are extremely insulin sensitive and live longer than their littermate controls. By contrast, the overexpression of GH and consequent enhancement of IGF-­‐1 action in bGH mice results in larger and leaner bodies, extreme insulin resistance, and shorter lifespan than littermate controls. We applied Western blot techniques on samples of gastrocnemius/soleus complex homogenate from each genotype, using a monoclonal antibody against the C-­‐terminus of human mstn protein, and a purified mstn peptide as control. We found no statistically significant difference in the levels of expression 28 of mstn in either the GHR-­‐/-­‐ or the bGH, compared to littermate controls, indicating that modifying the GH/IGF-­‐1 axis does not impact mstn levels, and also that changes in body composition in those animal models are not due directly to mstn. Muscle function in vivo was analyzed in the bGH mice vs. littermate controls with grip strength meter and rotarod behavioral assays. There was no significant difference in rotarod performance, indicating that the phenotypic changes associated with these genetic mutants does not affect balance, coordination, or endurance. However, grip strength assessment of rear limb pull force was significantly greater in littermate controls when normalized for body weight. Although changes in mstn were not evident in these models, the increased grip strength in controls contributes to existing knowledge (including models of mstn inhibition) that higher percentage of lean mass does not necessarily contribute to an increase in strength. Perhaps the difference can be attributed to the fact the littermate controls are only ~1/4 of their lifespan, while bGH mice are already ~halfway through their lifespan and have already begun to experience the effects of aging. Email: [email protected] 41 Marilyn Day Jennifer Round Slitrk1 is expressed in Rohon-­‐Beard sensory neurons of the developing zebrafish spinal cord. M. DAY, U. BAQAI, A. KANE, R. ARAS, J. ROUND Ursinus College Many of the cellular mechanisms that direct nervous system development are still unknown, and investigating them could be critical to treating nervous system disorders. The Slitrk protein family consists of six transmembrane proteins that are highly expressed in the central nervous system. This protein family has been associated with specific neuropsychiatric disorders such as OCD, anxiety, depression, Tourette syndrome, and schizophrenia. At the cellular level, Slitrks have been implicated in axon fasciculation, presynaptic differentiation, neuromuscular development, and neurite outgrowth. Slitrk1 mRNA is abundant in the developing spinal cord, but which cell types express Slitrk1 and what role it plays in spinal cord wiring are unknown. In this study, we employed immunohistochemistry to investigate Slitrk1 protein localization in the zebrafish spinal cord. We found that Sltirk1 is located in Rohon Beard neurons in early zebrafish development, and we observed Slitrk1 distribution patterns that suggest its presence in glial and/or neural crest cells. This work provides a foundation for future loss-­‐of-­‐
function studies to investigate a novel role for Slitrk1 in vertebrate spinal cord wiring. Our research will improve understanding of how the nervous system is assembled, and this information could potentially be used to help treat nervous system disorders. Email: [email protected] 42 Michael Shoats Ron Bayline The study of Notch-­‐Delta signaling pathway in Manduca sexta M. P. SHOATS, K. MCGOWEN, R. BAYLINE, Y. LAI Washington and Jefferson College During cell specification, cell-­‐cell interactions mediated by direct contact often regulate determination of cell fates. The Notch-­‐Delta signaling pathway involves transmembrane proteins that regulate cell fate determination in many cell types. For example, in Drosophila muscle development during embryogenesis, Notch-­‐Delta interactions regulate the determination of muscle founder cell fates. High levels of Notch expression leads to epidermal cell growth from the ectodermal precursor cells while low levels of Notch, and thus higher levels of Delta, result in differentiation of ectodermal cells into neural cells. During postembryonic development in Drosophila, myoblasts approach their targets and are maintained in a 29 semi-­‐differentiated state by continuous Notch activation. High levels of Notch during this time in development results in a lack of fused, multinucleated cells. The role of notch-­‐delta interactions during muscle developing in other insects has not been elucidated. In the moth Manduca sexta, muscles display patterns of development than that seen in Drosophila. Both nerve-­‐dependent and nerve-­‐independent muscle development can occur during adult development. Additionally, due to the size of the Manduca the Notch pathway may play a different role in postembryonic muscle development. This research investigates the role of Notch-­‐Delta signaling during Manduca sexta embryonic and postembryonic muscle development. In this project, we focus on the development of flight muscles and the tergosternal muscle, which display nerve-­‐dependent and nerve-­‐independent development, respectively. We hypothesize that Notch-­‐Delta interactions may exhibit different roles in specifying myoblast fates between these two types of muscles. We have identified a putative Notch gene from the Manduca genome database based on sequence homology with the Drosophila Notch gene. PCR primers sequences for the putative Notch gene were developed, and a PCR fragment was isolated from a Manduca embryonic cDNA library. The fragment was ligated into a plasmid vector and bacteria were transformed. The plasmids were then isolated, and the amplified fragment was sequenced to confirm its identity as a putative Notch homolog. Riboprobes developed from the PCR fragment were used for in situ hybridization to characterize expression in embryos. We predict that Notch-­‐Delta signaling will be detected during M. sexta muscle development in the embryo. The levels of expression during differing stages can be an important indicator of the role that Notch plays during different types of muscle development. Understanding the role of Notch-­‐Delta signaling during M. sexta muscle development will help elucidate the evolution of muscle patterning in insects. Email: [email protected] 43.1 Ranran Li Elizabeth Weaver On how parameter variations influence bursting activity R. LI, A. DOLOC-­‐MIHU, R. CALABRESE Agnes Scott College Recent experimental and computational studies suggest that linearly correlated sets of parameters (intrinsic and synaptic properties of neurons) allow central pattern-­‐generating (CPGs) networks to produce and maintain their rhythmic activity regardless of changing internal and external conditions. Within an existing database of the half-­‐center oscillator (HCO; Hill et al, 2001) model instances of the leech heartbeat CPG, a recent computational study suggests that there may be non-­‐linear relationships between parameters for HCO instances (Doloc-­‐Mihu et al., 2014). In order to further understand the non-­‐linear relationships between parameters for HCO instances, we picked specific subgroups of data from the HCOs. The first set is a subgroup from the realistic HCO group, and includes all the 8 members of the gBarh families of the realistic HCO group. The second set is a subgroup from the unbalanced HCO group, and includes those instances, which become tonically spiking isolated neurons when there is no synapse present between the two neurons. We applied the Principal Component Analysis (PCA) method to both sets. PCA did not identify any linear correlation between parameters in these two subgroups. However, visualizations of these two subgroups in a reduced space suggested that there might be non-­‐linear correlations between some parameters. In light of the sensitivity of period to the changes in gBarh current revealed by recent study, we visualized the relations of all the parameters with each characteristic (period, spiking frequency and duty cycle) in both subgroups. We found that for the first set, spiking frequency shows sensitivity to the changes in each gBarEleak, gBarP, and gBarCaS currents. Visualization also verified the sensitivity of period to the changes in gBarh current. 30 43.2 Arille Lewis Elizabeth Weaver Upregulation of Ataxin-­‐2 in BLOC-­‐1 deficient cells A. LEWIS, A. BURCH, S. ZLATIC, V. FAUNDEZ Emory University Biogenesis of Lysosome-­‐related Organelles Complex -­‐1 (BLOC-­‐1) is composed eight protein subunits involved in endocytosis. BLOC-­‐1 is required for targeting specific proteins to synaptic vesicles and lysosome-­‐related organelles from endosomes. One subunit, Dysbindin (dystrobrevin-­‐binding-­‐1), is encoded by the DTNBP1 gene, which is a gene associated with susceptibility to schizophrenia. The function of dysbindin and BLOC-­‐1 remains to be elucidated. The precise molecular pathways at the synapse downstream of BLOC-­‐1 and dysbindin responsible for these changes are the focus of my studies. I determined the cellular levels of these proteins in BLOC-­‐1 deficient Pallidin knockdown SH-­‐SY5Y cells. ATXN2, which causes Spinocerebellar Ataxia-­‐2 (SCA2) in humans, plays a role in mRNA translation and synaptic plasticity. My data show that VAMP7 abundance decreases and reciprocally ATXN2 abundance increases in BLOC-­‐1 deficient SH-­‐SY5Y cells. Data here suggest the possibility that ATXN2 functionally interact with BLOC-­‐1 subunit proteins and may affect RNA metabolism associated with endosomes, and synaptic vesicle function. 44 Caleb Glassman Nancy Kleckner AMPA-­‐like receptor subunits localized to glutamate-­‐responsive feeding motor neurons in the pond snail, Helisoma trivolvis C. GLASSMAN, J. QIN, N.W. KLECKNER Bates College Glutamate is a common neurotransmitter in both vertebrate and invertebrate nervous systems. In the pond snails Helisoma trivolvis and Lymnaea stagnalis, glutamate released onto buccal ganglion neurons patterns motor output to the buccal mass to promote the standard pattern of feeding. Glutamate excites motor neurons activating phase 2 of feeding and inhibits motor neurons in phases 1 and 3 of the tripartite feeding pattern, but the receptors mediating these effects have not been well characterized. Two receptors have been cloned from Helisoma central nervous system: one (Hel-­‐GluR1) has a high degree of sequence similarity with an AMPA-­‐like receptor from Lymnaea, Lym-­‐eGluR1; a second, partial receptor (Hel-­‐GluR7) has a high degree of sequence similarity with a KA-­‐like receptor from Aplysia californica, Apl-­‐GluR7. The purpose of this study was to determine the specific neural location of these excitatory glutamate receptors within the known Helisoma feeding circuitry. It was hypothesized that phase 2 motorneurons would contain the AMPA-­‐like receptor Hel-­‐GluR1, as those neurons are responsive to KA in a CNQX-­‐dependent manner, and that phase 3 motorneurons would not, as they respond to glutamate with hyperpolarization. To test this hypothesis, an antibody that recognizes rat GluR2 and 3 subunits was used to localize AMPA-­‐
like receptor subunits to neurons within the Helisoma buccal ganglia, and primers were designed to amplify portions of Hel-­‐GluR1 with single-­‐cell PCR. Sequences were amplified from cDNA reversed transcribed from mRNA isolates of individual motor neurons B27 and B19, corresponding to phases 2 and 3, respectively, of the Helisoma feeding central pattern generator. Surprisingly, with both techniques, AMPA-­‐like receptors were localized to both B27 and B19 neurons. Hel-­‐GluR1 was not identified in neuron B5, which innervates the esophagus. Additional experiments are underway to determine which buccal neurons contain the Hel-­‐GluR7 receptor subunit. It is anticipated that, since buccal A cluster neurons that modulate the feeding CPG are immunoreactive to an antibody made against a rat GluR5 sequence, these neurons might contain this KA receptor-­‐like subunit. Understanding the number and localization of glutamate receptor subunits in buccal neurons will improve our understanding of how glutamate patterns the feeding motor output. This project was supported by grants from the National Center for Research Resources (5P20RR016463-­‐12) and the National Institute of General Medical Sciences (8 P20 GM103423-­‐
12) from the National Institutes of Health. 31 Email: [email protected] 45 Thomas Voigt R.J. Howard Modularity of alcohol binding sites in pentameric ligand-­‐gated ion channels T.B. VOIGT, H.A. BRAUN, D.E. MANSON, K.R. ELLIS, S. HORANI, G.E. JOYCE, J.-­‐A. M. FACEY, A.E. PLANTE, E.P. STATER, J.R. TRUDELL, R.A. HARRIS, R.J. HOWARD Skidmore College Alcohol’s effects on the brain and body are known to be associated with a specific subset of pentameric ligand-­‐gated ion channels. For example, GABA(A) and glycine receptors are potentiated by alcohols as well as general anesthetics. Although the structures of GABA(A) and glycine receptors have yet to be determined at atomic resolution, model proteins may facilitate their structural analysis. The bacterial homolog GLIC is largely insensitive to ethanol; however, we previously showed that mutating the 14’ position in GLIC from phenylalanine to alanine resulted in potentiation by pharmacologically relevant concentrations of alcohol. Here, we quantified the effects of modulators on GLIC chimeras containing substitutions of various lengths from homologous domains of GABA(A) and glycine receptors. For example, a chimera containing the glycine receptor transmembrane domain recapitulated glycine receptor potentiation by ethanol under equivalent experimental conditions. We used more localized substitutions at alcohol-­‐binding residues identified in a recent co-­‐crystal structure of GLIC to identify necessary determinants of alcohol modulation. In particular, a single substitution at the 14’ position to the equivalent residue in either alpha or beta GABA(A) receptor subunits conferred GABA(A) receptor-­‐like alcohol effects. Our results support the existence of a conserved binding site for allosteric modulators, such as alcohols, across ligand-­‐gated ion channels from multiple species. This work was supported by the National Institutes of Health National Institute on Alcohol Abuse and Alcoholism and by the Skidmore College Summer Faculty/Student Research Program. Email: [email protected] 46.1 Asia Ayabe Jonathan King Impact of short-­‐term voluntary and forced exercise on long-­‐term potentiation in the rat hippocampus. A. AYABE, K. GUAN, J. KING Pomona College Forced and voluntary exercise regimens have been well established as separate means of promoting cognitive function. Prior studies that directly compare the effects of forced and voluntary exercise have shown that both forms of exercise increase levels of brain-­‐derived neurotrophic factor, but effects on spatial memory tasks and synaptic plasticity have been variable. The current investigation compares the effects of forced and voluntary exercise on hippocampal synaptic plasticity as determined by long-­‐term potentiation (LTP). Our exercise paradigm was designed to minimize the impact of stress and pain, and standardized equipment and housing environments. The voluntary exercise group had free access to freely rotating running wheels and the forced exercise group used a motorized running wheel. Additionally, the voluntary exercise group ran quickly, and for shorter periods of time, when compared to forced exercisers. Our results show a significant increase in hippocampal LTP for the forced, but not voluntary, rats in comparison to the control. These findings suggest that periods of low intensity, longer duration exercise may be more beneficial to synaptic plasticity than high intensity, short duration regimens. Overall, voluntary and forced running are two distinct models that produce different effects on LTP, and should be treated as such. Email: [email protected] 32 46.2 Yuni Kay Jonathan King Ginsenoside metabolite Compound K enhances hippocampal LTP but does not protect or repair Aβ-­‐induced deficits on LTP Y. KAY, A. TRIBBY, J. KING Pomona College Amyloid beta peptides are implicated in cognitive deficits associated with Alzheimer's disease. Buildup of Aβ deposits in the hippocampus is thought to contribute to cognitive decline. Research suggests that the active components of ginseng roots called ginsenosides, such as Rb1, may ameliorate the effects of some forms of neurodegeneration. Additionally, Rb1 is metabolized in the body and Compound K (ComK), its final metabolite, is present at the highest concentrations in the blood. Preliminary data in the lab has indicated ComK's potential ability to counteract the detriments caused by Aβ25-­‐35. This experiment further investigated ComK's role in LTP modulation and studied its potential mechanism by investigating variable temporal application. Our results show that compared to the control, ComK facilitates LTP and Aβ attenuates LTP. However, ComK does not protect against attenuation caused by Aβ. Furthermore, ComK attenuates LTP if applied after Aβ perfusion. Thus, while 5nM ComK increases LTP compared to the control, it is not an effective treatment against Aβ-­‐induced impairments in hippocampal synaptic plasticity. Email: [email protected] 47 Scott Newton Jeff Edwards Ventral tegmental area dopamine and GABA neurons: physiological properties and expression of mRNA for endocannabinoid biosynthetic enzymes and type I mGluRs C.B. MERRILL, L.N. FRIEND, S.T. NEWTON, Z.H. HOPKINS, J.G. EDWARDS Brigham Young University The ventral tegmental area (VTA) is known to control the processing of rewarding and addictive behaviors. The VTA contains dopamine (DA) cells, which release DA to downstream targets in response to rewarding stimuli, and GABA cells, which modulate DA cell activity. Therefore, both cell types are involved in associative reward learning. Synaptic plasticity plays an important role in adaptive reward signaling within the VTA. Endocannabinoids can mediate or modulate synaptic plasticity at several synapses within the reward circuit. However, the source of endocannabinoids within the VTA is not well understood. Therefore, our goal was to describe the distribution of endocannabinoid biosynthetic enzyme mRNA within VTA neurons. We extracted single VTA neurons via whole cell patch clamp and used single-­‐
cell real-­‐time quantitative PCR to identify DA and GABA neurons based on mRNA expression of cell-­‐type specific targets. DA neurons were identified by the presence of tyrosine hydroxylase and DA transporter mRNA, while GABA neurons expressed GAD65 and GAD67 mRNA. Additionally, electrophysiological properties such as action potential frequency and sag potential amplitude were examined between the two cell types. Concurrent with established observations, slower firing frequencies and larger Ih potentials were observed in DAergic neurons, however overlap was identified between these two cell types. VTA neurons were then probed for endocannabinoid/eicosanoid biosynthetic enzyme mRNA, such as N-­‐acyl-­‐phosphatidylethanolamine-­‐specific phospholipase D (NAPE-­‐PLD), diacylglycerol lipase α (DAGLα), and 12-­‐lipoxygenase. We also probed for type I metabotropic glutamate receptor (mGluR) mRNA, as endocannabinoid synthesis requires mGluR activation in many cases. Our data demonstrate that endocannabinoid biosynthetic enzyme mRNA is expressed in both DAergic and GABAergic cells with concurrent expression of type I mGluRs. Next, to ensure mRNA expression was representative of protein content, slices were stained using immunohistochemistry for GAD67, DAGLα, NAPE-­‐PLD and type I 33 mGluRs. Positive labeling for these targets was observed in VTA neurons, supporting our RT-­‐PCR results. Collectively, these data suggest DAergic and GABAergic cells of the VTA have the capability to produce endocannabinoids and potentially alter synaptic plasticity involved in reward and addiction. Email: [email protected] 48 Garrick Salois Charles L. Weaver Do enriched environments alter neuroplasticity in the rat dentate gyrus? A study of the effects of environmental complexity on the anatomy of the hippocampal neurogenic niche. G.J. SALOIS, J.S. SMITH The Brain Research Laboratory, Saginaw Valley State University, University Center, MI A typical laboratory housing environment for rats used in neurological studies involves no social interaction and limited sensory stimulation. In contrast, an enriched environment involves housing animals in groups, providing room for exercise and exploration, and the regular introduction of novel stimuli in the form of species-­‐appropriate toys. In order to accommodate the more complex sensory inputs and motor behaviors that result from enriched environments, the brains of research animals undergo anatomical changes broadly categorized as neuroplasticity. One of the effects of enriched environments on the physiology of the brain is the stimulation of neurogenesis in the dentate gyrus of the hippocampus. To date, limited research has been conducted to assess morphological changes in response to enriched environments amongst cells participating in the neurogenic niche. This study utilized a technique called SeeDB, which renders large sections of neural tissue transparent, obviating the need for serial sectioning when imaging large anatomical regions. SeeDB was performed on brain samples of animals reared in either enriched environments or standard laboratory housing. Another technique, immunohistochemistry, allows specific cellular structures to be labeled with fluorescent antibodies. By combining these techniques, 250 µm sections of rat dentate gyrus were imaged in three dimensions using a confocal microscope. Anti-­‐doublecortin was used to label the complete dendritic arbor and cell body of immature neurons in the dentate gyrus. Anti-­‐GFAP was used to assess changes in astrocytes, a glial cell critical in the maturation and support of newborn neurons. In addition, anti-­‐NeuN was used to assess the number of neurons in the hippocampus. It is hypothesized that animals housed in enriched environments will have a greater number of doublecortin-­‐positive neurons in the dentate gyrus than animals housed in standard environments. Furthermore, these neurons will have more complex dendrites in terms of length and number of branches. Animals housed in enriched environments are also predicted to have a greater number of astrocytes to accommodate the increased resource demands of stimulated neurogenesis. These environmentally-­‐mediated anatomical changes may represent a closer approximation of normal physiological functioning in the hippocampus compared to animals housed in standard laboratory housing, which may have implications in the effective use of rats as a research model. Email: [email protected] 49 Kathleen Luckett Bruce Johnson Remodeling of the guinea pig intrinsic cardiac nervous system with chronic heart disease K.A. LUCKETT, J.C. HARDWICK Cornell University Chronic heart disease, such as myocardial infarction (MI), produces remodeling of the autonomic nervous system and induces an increase in sympathetic output, as well as remodeling of the intrinsic cardiac nervous (ICN) system located within the heart. Sympathetic fibers innervate the parasympathetic 34 intracardiac neurons of the ICN and thus, increased sympathetic activity could lead to altered function within the cardiac plexus. To examine this possibility, we looked at the responses of parasympathetic cardiac neurons to the application of sympathetic neurotransmitters (norepinephrine (NE) and neuropeptide Y (NPY)). MI was surgically-­‐induced in guinea pigs by ligating the left anterior descending coronary artery on the surface of the left ventricle. After a four week recovery period, whole mounts of the intrinsic cardiac plexus were prepared for intracellular voltage recording. NE and NPY were applied by local pressure ejection. In addition, fiber bundles innervating the neurons of interest were stimulated with an extracellular focal electrode to monitor synaptic transmission. In control animals, NPY produced a small increase in neuronal excitability, while NE had no significant effects on excitability. Simultaneous application of both substances did not differ from NPY alone. In animals with MI, both NE and NPY produced a small increase in neuronal excitability, while simultaneous application significantly increased excitability. This combinatory effect of simultaneous NE and NPY application is being investigated further in both control and MI animals. In addition, we found that the combined application of NE and NPY in the MI animals also significantly increased the ability of presynaptic inputs to produce postsynaptic action potentials at high frequencies (20Hz). Combined, this data suggests that the MI-­‐induced remodeling of the sympathetic nervous system also increases the parasympathetic responses, perhaps as a mechanism to compensate for the excessive sympathetic activity. Email: [email protected] 50.1 Cristina Risi Darlene Mitrano Analysis of the localization of the µ-­‐opioid receptor in relation to cocaine and morphine withdrawal C. RISI, M. MABALOT, E. COMITA, D. MITRANO Christopher Newport University Drug and alcohol addictions are a continuing societal issue due to addicts attempting to avoid withdrawal symptoms and concurrent relapse. It is thought that relapse may be caused by a receptor’s specific response to a drug, which in turn causes a change in neuronal functioning. The µ-­‐opioid receptor, which can be found in the nucleus accumbens, plays an important role in the drug reward pathway. Recent studies have demonstrated that stimulants, such as cocaine, and depressants, such as morphine, may alter the endogenous opioid system. Morphine directly acts on the µ-­‐opioid receptor within the nucleus accumbens and throughout the brain, while other studies have shown that cocaine acts indirectly on the opioid system, for example, by altering opioid modulation of GABA. Additionally both drugs act on the brain’s reward system by enhancing dopamine levels. The goal of this study is to investigate not only the location of the µ-­‐opioid receptor, but to explore the role and alterations of this receptor after the withdrawal process in rodents has occurred. Fifteen rats were treated for seven days with morphine, cocaine, or saline. The rats were left untreated for seven days to mimic withdrawal. After the withdrawal period, the brain tissue was obtained by transcardial perfusion. Ensuing fixation, the brains were sliced into 60 µm sections using a vibrating microtome. Using immunocytochemistry the tissue containing the nucleus accumbens was stained for the µ-­‐opioid receptor and processed for both light and electron microscopy (EM). For observation at the EM, immunoperoxidase is used to ascertain which neural elements contain the receptor, while the immunogold technique is used to determine the relative location within the neuronal element that the receptor is found. We hypothesize that the µ-­‐opioid receptor will change localization in the morphine and cocaine treated rats and not in the saline treated rats, especially after examination using the immunogold technique. Currently immunoperoxidase data is under analysis. Overall, it is the hope that this study will provide a basis for understanding the µ-­‐opioid receptor’s role in cocaine and morphine addiction and for the development of new treatment options for withdrawal periods to avoid relapse. Email: [email protected] 35 50.2 Kelsey Jackson Darlene Mitrano Colocalization of D1-­‐dopamine receptors and α1b-­‐adrenergic receptors in the rat nucleus accumbens K. JACKSON, A. SEELEY, D. MITRANO Christopher Newport Unversity Areas of the brain collectively known as the reward system consist of the prefrontal cortex (PFC) and the nucleus accumbens (NAc). They are important for various processes, including attention, impulse control and feelings associated with pleasure and reward. Normal functioning of these brain areas is impaired in disorders like drug addiction and attention deficit/hyperactivity disorder (ADHD). Neurotransmitters such as dopamine and norepinephrine, and their receptors have shown to play a role in regulating these processes. The goal of this study is to further explore the location, role and interactions of norepinephrine and dopamine receptors in the NAc. Previous studies have shown a high degree of colocalization of these receptors within areas of the rodent PFC. In this study, tissue containingthe shell and core of the NAc was stained using immunocytochemical techniques and examined using electron microscopy (EM). Specifically, tissue was labeled with antibodies for the D1-­‐dopamine receptor (D1R), the alpha1b-­‐
adrenergic receptor (α1bAR) or both, using immunoperoxidase or immunogold to reveal receptor localization. Preliminary data of immunoperoxidase staining has shown a high density of D1Rs localized primarily in post-­‐synaptic elements, such as dendrites and spines. Tissue labeled for the α1bAR is still be analyzed, as well as single labeled tissue using the immunogold technique. Preliminary observations of double labeled tissue shows that about 10-­‐20% of D1R containing elements also have labeling for the α1bAR, which varies significantly from results in the PFC. Future studies also include looking at these receptors in the PFC and NAc following acute and chronic cocaine treatment. It is the hope that understanding receptor localization and dynamics after drug exposure will lay the foundation for understanding why these drugs are so addictive and to develop better treatments for ADHD and psychostimulant addiction. Email: [email protected] 51 Tae-­‐Hyun Sakong James L Roberts Analyzing the effect of Estradiol on astrocytic Apo E production T.H. SAKONG, J. ATONE, A. LEEK, J.L. ROBERTS Trinity University Apolipoprotein E (apoE) is a lipid transport protein that mediates cholesterol transport and clearance within the central nervous system (CNS). The dynamic exchange of apoE between neuronal and non-­‐
neuronal cells indicates the fundamental importance of apoE mediated cholesterol transport. Although the e4 allele of apoE is a major genetic risk factor for Alzhemier’s Disease (AD), the mechanism of apoE mediated cholesterol synthesis, transport, and/or secretion remains to be elucidated. Using primary cultures of midbrain astrocytes from 4, 12, 17, and 28 month old mice, prelimnary studies using indicate that in astrocytes, apoE expression changes as a function of age. RT-­‐qPCR analyses show that ApoE expression is low at younger (4M) and older ages (28M), but the highest at the middle ages (17M). Interestingly, the inductive effects of estrogen treatment on apoE expression were the greatest at older ages. To couple the gene expression analyses, cytoplasmic apoE production and secretion was quantified using an ELISA assay. Furthermore, our lab also investigated the effect of 2-­‐MeSADP to investigate the role of purinergic signaling in apoE production in astrocytes. In the CNS, 2-­‐MeSADP is known to bind to purinergic receptors and induce cell proliferation through the mitogen-­‐activated protein kinase (MAPK) pathway. Because estradiol mediates signaling cascades through the MAPK pathway, this study aims to see if 2-­‐MeSADP induces the similary effects on apoE expression, production, and or secretion. Furthermore, neurons grown in combinatorial conditions including with astrocytes, estradiol treatment, 36 and oxidative stress (H2O2) were stained for apoE and were imaged using Nikon confocal microscopy. Though the results are not yet conclusive, 2-­‐MeSADP has so far resulted in a slight increase in apoE. Email: [email protected] 52 Jennifer Rojas Kathryn Reissner The effects of riluzole on cue-­‐ and cocaine-­‐primed reinstatement of cocaine seeking J. ROJAS, A. AURIEMMA, K.L. HEALEY, K. REISSNER University of North Carolina-­‐Chapel Hill Chronic drug use has been shown to suppress the expression and activity of glutamate transporters and thereby disrupt glutamate homeostasis. Disruption of glutamate homeostasis in the nucleus accumbens has been established as an important cellular mechanism regulating drug-­‐seeking behavior; hence, interventions that restore the expression of glutamate transporters and glutamate homeostasis in this region have recently become considerations for therapeutic targets of addiction. Riluzole, an FDA-­‐
approved treatment for amyotrophic lateral sclerosis, has been previously demonstrated to be a positive regulator of glutamate transporter GLT-­‐1. Hence, we hypothesize that Riluzole, when administered chronically after cocaine self-­‐administration, will decrease cocaine-­‐seeking behavior in rats. To test this hypothesis, rats were trained to self-­‐administer cocaine for ten days via an intravenous catheter. During each cocaine infusion, a light-­‐tone compound stimulus was presented. Rats were then given nineteen days of either daily Riluzole or vehicle treatment i.p., thirty minutes prior to daily extinction sessions in which both drug and light-­‐tone cues were absent. Subsequently, all rats underwent two reinstatement tests. The first was a cue-­‐primed reinstatement test during which they were presented with the contingent light-­‐
tone stimulus. After 3 additional days of extinction and treatment, rats received a cocaine-­‐primed reinstatement test, before which they received Riluzole treatment followed by administration of cocaine (10 mg/kg, i.p.). Results showed that Riluzole (4 mg/kg) had a trend toward an effect on cue-­‐primed reinstatement, but was without effect on cocaine-­‐primed reinstatement at any dosage. In addition, Riluzole had no effect at any dose on cue-­‐primed reinstatement of sucrose seeking. Subsequent experiments are ongoing to determine whether the effect on cue-­‐primed reinstatement will prove significant with greater statistical power. Email: [email protected] 53 Paul Austin Thomas Heinbockel Interaction of two neurotransmitter systems in olfactory neuronal circuits. P.T. AUSTIN, B.T. OLIVER, T. HEINBOCKEL University of the District of Columbia (PTA), Howard University College of Medicine (BTO, TH) PURPOSE: Protecting neurons in the brain against excitotoxicity and neuronal death is critical during traumatic experiences such as stroke. Brain-­‐produced endogenous cannabinoids (eCBs) are prime candidates for a self-­‐defense mechanism because of the known neuroprotective effects of exogenously applied cannabinoids. Δ9-­‐tetrahydrocannabinol (THC), the bioactive ingredient of marijuana, activates cannabinoid receptors (CB1R) in the brain in the same manner as eCBs. Main olfactory bulb (MOB) neurons express high levels of CB1R. Glutamate acts on ionotropic as well as metabotropic glutamate receptors (mGluRs) in the brain. In the MOB, mGluRs (mGluR1) are expressed at high levels by mitral cells. We study how these two neuromodulator systems interact to regulate activity of mitral cells by potentially exerting neuroprotective or neurotoxic effects. METHODS: In mouse brain slices, we used whole-­‐cell patch-­‐clamp recordings to study how CB1R and mGluRs regulate mitral cell activity. RESULTS: Mitral cells respond with potent membrane potential depolarization and increased action potential firing in response to activation by group I mGluR agonists. mGluR1 antagonists block this effect and reduce the firing activity. CB1R agonists evoke increased action potential firing, while a CB1R antagonist reduces 37 firing. Novel effects are observed when receptor blockers for both CB1 and mGlu1 are combined. Rather than reducing mitral cell activity as observed individually, the combination of both antagonists results in excessive action potential firing leading to cell death. CONCLUSION: CB1Rs and mGluRs regulate mitral cell activity. Inhibition of both receptor systems leads to neuronal overstimulation of mitral cells, but could have neuroprotective effects when both systems are activated. SUPPORT: PTA is supported through The Advanced Research Training Corps: A Novel Initiative for URM Students, NIH-­‐NIGMS [GM101997]. BTO is supported through the Howard University College of Medicine Medical Student Summer Research Program, 2014. TH is supported by NIH [MD007597] and NSF [IOS-­‐1355034]. Email: [email protected] 54.1 McKenna Redwine William Blaker Nerve growth factor expression in the septum and the dentate gyrus during lesion-­‐
induced septodentate axonal sprouting M.R. REDWINE, W. BLAKER Furman University In this study, changes that occurred during septodentate axonal sprouting were examined in adult rat brains. To initiate this sprouting, a stereotaxic instrument was used to electrically lesion the entorhinal cortex on one side of each brain, leaving the other as a control side. The rat’s brains were dissected 15 days post-­‐lesion and hippocampal tissue sections were stained for acetylcholinesterase (AChE) to confirm if sprouting had indeed occurred. RNA was also isolated and preserved from both the septum and the dentate gyrus so that real-­‐time reverse transcriptase PCR could be used to determine the amount of nerve growth factor (NGF) expression in each brain region, respective to one another as well as to each corresponding control. At the conclusion of this study, axonal sprouting was confirmed in the hippocampus, while NGF expression was up-­‐regulated in the septum and unchanged in the dentate gyrus. Further studies should focus on why NGF is up regulated in the cell body region of the sprouting pathway, rather than in the terminal region where the sprouting actually occurs. Email: [email protected] 54.2 Manuel Hazim William Blaker CSF and neuropsychological correlates of visual hallucination in dementia with lewy bodies M.G. HAZIM, W.T. HU, C.M. HALES, J.J. LAH, A.I. LEVEY Furman University Objective: To identify clinical features of dementia with Lewy bodies (DLB) least likely associated with Alzheimer's disease pathology, and to determine whether it is associated with a unique neuropsychological profile. Methods: Clinical records of 98 patients given the diagnosis of DLB at a specialty cognitive and behavioral neurology clinic in a tertiary referral center were retrospectively reviewed for core, suggestive, and supportive features of DLB as well as demographic variables, cerebrospinal fluid (CSF) Alzheimer's biomarkers, and longitudinal neuropsychological analyses. Results: Core, suggestive, and supportive features were common in this cohort, with 69% and 39% of patients assigned the diagnosis of probable DLB and possible DLB fulfilling consensus criteria for probable DLB. 26 of 98 clinically diagnosed DLB patients had CSF Alzheimer's biomarker analysis, and visual hallucination was the only feature not associated with CSF suggestive of Alzheimer's disease. 42 of 98 patients had longitudinal neuropsychological analyses, and patients with visual hallucinations had worse baseline executive functions but slower longitudinal decline in executive functions than patients without visual hallucinations. Conclusion: Visual hallucination in clinically diagnosed DLB is associated with CSF 38 biomarkers consistent with a non-­‐AD disorder and a unique longitudinal neuropsychological profile. DLB patients with visual hallucinations can be considered a unique DLB endophenotype for future biomarker discovery and validation. Email: [email protected] 55 Dana Sautter Timothy E. Koeltzow Repeated pairing of cocaine with an anxiogenic environment impairs the acquisition of conditioned place preference, but promotes behavioral sensitization in chronically food-­‐
restricted rats. D.M. SAUTTER, G.J. BAKER, J.M. SMITH, L.A. THOMPSON, T.E. KOELTZOW Bradley University Stress is known to increase the response to cocaine in preclinical models of addiction, including behavioral sensitization, conditioned place preference (CPP), and operant cocaine self-­‐administration (Miczek et al., 2011; Piazza & LeMoal, 1998). In addition, stress is sufficient to reinstate extinguished cocaine-­‐seeking behavior, suggesting that stressors represent a significant risk factor for cocaine abuse and relapse (Back et al., 2010; Erb, 2009). In rodents, food-­‐restriction (FR) represents a chronic mild stressor that has been shown to enhance a rodent’s response to a subsequent cocaine challenge. The purpose of the present experiment was to assess the impact of repeatedly pairing cocaine (7.5 mg/kg/day) with an anxiogenic environment among rats undergoing chronic FR compared to free-­‐feeding (FF) rats. Our results indicate that FR promotes hyperactivity at baseline in a Light/Dark (Lt/Dk) activity chamber sufficient to counter the behaviorally-­‐inhibiting effects of the anxiogenic light side of the compartment (F (1,34) = 22.18, p < 0.001). Consequently, FR rats spent statistically significantly more time exploring the Lt side of the chamber (F (1,34) = 12.63, p < 0.001), perhaps an artifact of food foraging behavior or an enhanced dopamine response to the novel test environment. Following 8 days of conditioned place preference training, whereby all rats were administered cocaine on either the Lt or Dk compartment on alternating days (FF/Dk: n=9: FF/Lt: n=9; FR/Dr: n=9: FR/Lt: n=9), the aversive influence of the light side of the chamber remained the most powerful determinant of behavior, countering the ability of cocaine to establish robust CPP. Nevertheless, cocaine pairing clearly had a behavioral effect as it was found that, among rats in which cocaine was paired with the anxiogenic Lt side of the chamber, statistically significantly less locomotor activity was observed when rats were placed in an anxiogenic open field apparatus, an effect that was observed among both FF and FR rats (t (34) = 1.99, p<0.05). These findings suggest conditioning mechanisms can be sufficiently powerful so as to counter the conditioned locomotor-­‐activating effects of cocaine. Lastly, consistent with previous reports, it was found that FR resulted in a sensitized locomotor response to cocaine (15 mg/kg) when rats were challenged in a novel test environment (F (1,34) = 6.21, p < 0.05). Control group data collection remains in progress. Email: [email protected] 56 Kathryn-­‐Mary Wakim DJ Brasier Effect of adversity on addiction related circuitry K.M. WAKIM, J. FEDORKO, V. PARIYADATH, E.A. STEIN Carnegie Mellon University & National Institute on Drug Abuse Childhood exposure to adverse life events is a significant risk factor for the later development of substance abuse (Oswald et al, 2013). Previous studies disagree as to how precisely IC activation is affected by adversity, with some arguing that adversity increases IC activation by over stimulating prefrontal-­‐limbic-­‐striatal circuits (Seo et al, 2014) and others arguing that IC activation decreases during reward processing tasks as a function of the higher rates of depression often reported by maltreated 39 individuals. No previous studies have investigated the relationship between adversity and connectivity within addiction-­‐related circuits. The present study uses diffusor tensor imaging (DTI) to elucidate the effect of adversity on local connectivity within the internal capsule, a white matter tract implicated in addiction circuitry (Buckholtz et al, 2010). We found a significant effect of childhood trauma (as measured by the Childhood Trauma Questionnaire) on local internal internal capsule as well as midbrain regions near the SNR and VTA, both of which contain a high volume of afferent projections to the striatum. Our results agree with recent studies suggesting that the elevated rates of depression in maltreated individuals often co-­‐occur with decreased striatal activation during reward processing tasks (Dillon et al, 2009) and suggest that connectivity decreases as a function of adversity. Support: Summer Internship Program in Biomedical Research at NIH Email: [email protected] 57 Megan Menosky Julien Rossignol Use of induced-­‐pluripotent stem cells as a potential therapy for Parkinson’s disease. M. MENOSKY, S. DECKER, G. SHALL, R. WELCHKO, X. LEVEQUE, J. ROSSIGNOL, G. DUNBAR Central Michigan University Parkinson’s disease is a late, continuous and progressive neurodegenerative disorder caused by the loss of dopaminergic neurons in the substantia nigra. Currently, the treatments available are only palliative and cannot cure the disease. Induced pluripotent stem cells (iPSCs) have considerable potential as a treatment for PD because they can be derived from adult tissue and are able to differentiate into all cells, including DA neurons, which are depleted in PD. In this study, we utilized sonic hedgehog (Shh), fibroblast growth factor 8 (FGF8) and glial derived neurotrophic factor (GDNF), and compared the ability of these factors with that of purmorphamine (Pur), a cost effective Shh agonist, to induce the differentiation of iPSCs into DA neurons. We found that treatment with both Shh and Pur successfully generated DA neurons from iPSCs. These cells were then transplanted into rats given injections of the 6-­‐hydroxydopamine rat (6-­‐
OHDA) to test the efficacy of these differentiated iPSCs to counteract motor deficits in this model of PD. In the future we will investigate if the transplanted cells that were cultured with Shh or Pur are able to integrate in the host brain and help to counteract the motor deficits caused by the 6-­‐OHDA injections. These results support other studies which indicate that iPSCs hold the potential to be a cellular therapy for the treatment of PD. Email: [email protected] 58.1 Karen Monuszko Julio J. Ramirez The influence of septodentate stimulation on long-­‐term potentiation in the crossed temporodentate pathway 12 days after entorhinal cortex lesion K.A. MONUSZKO, J.S. ISAAC, N. UPRIGHT, M. MOSES-­‐HAMPTON, J.J. RAMIREZ Davidson College The hippocampus receives strong innervation from the entorhinal cortex, a structure that experiences severe neuronal loss during Alzheimer’s disease. A unilateral entorhinal cortex lesion damages the main projection from the entorhinal cortex to the hippocampus. To compensate, two other pathways, the crossed temporodentate (CTD) and septodentate (SD), sprout to elicit a response from the granule cells in the dentate gyrus of the hippocampus. The strength of the neuronal response can be measured in the form of extracellular field potentials (EPSPs) in the dentate gyrus. Paired-­‐pulse stimulation of the SD and CTD may affect EPSP amplitude or slope in comparison with unpaired stimulation of the CTD. Long-­‐term potentiation (LTP), a potential mechanism of learning and memory, can be induced into the CTD to observe any changes in the characteristics of the EPSPs. We sought to determine how LTP induction 40 affected the paired-­‐pulse interaction 12 days after a unilateral entorhinal cortex lesion. Rats received either a unilateral lesion of the right entorhinal cortex or a sham operation. Twelve days later, EPSPs in the dentate gyrus were recorded in response to both paired-­‐pulse and unpaired stimulation at varied intensities both before and after LTP induction. Histology assessed the extent of the lesion and the accuracy of electrode placement. The slope, representing the speed of the neuronal response, and amplitude, representative of the approximate number of neurons firing, of EPSP waveforms were measured. These variables were compared between pre-­‐ and post-­‐tetany, sham and lesion cases, and paired and unpaired responses. This work is still in progress, but preliminary results suggest that amplitude is unaffected by paired-­‐pulse stimulation or by LTP induction. However, the lesion of the entorhinal cortex, in comparison to the sham operation, did lead to increased amplitude of the EPSP waveform after both paired and unpaired stimulation. Email: [email protected] 58.2 Sarah Hamitlon Julio J. Ramirez Septal facilitation of long-­‐term potentiation in the perforant path in rats S.E. HAMILTON, M. MOSES-­‐HAMPTON, J.J. RAMIREZ Davidson College Within the hippocampus, experiences are consolidated into memories with the help of the numerous afferents and efferents of the hippocampal region. Two pathways that contribute to this memory formation are the perforant pathway, a glutamatergic pathway arising from the entorhinal cortex and projecting to the ipsilateral hippocampus, and the septodentate pathway (SD), a primarily cholinergic pathway projecting bilaterally to the hippocampus. Long-­‐term potentiation (LTP) is a major mechanism that helps in memory formation and is seen as the neurobiological correlate of learning. LTP is an enduring increase in synaptic efficacy following tetany, a series of high frequency stimulations. This mechanism has been exhibited greatly within the perforant pathway, one of the largest afferents to the hippocampus. The role of the septum and SD within the workings of the hippocampus has not been fully determined, especially in regards to perforant pathway LTP. This study sought to expound upon the role of the SD within the hippocampus through the electrophysiological examination of its influence in the LTP abilities of the perforant pathway. The functioning of the septal and perforant pathway interaction was determined using a paired-­‐pulse paradigm with conditioning SD stimulation and pre-­‐ and post-­‐tetany (induction of LTP) comparisons. With conditioning SD stimulation, the response of the perforant pathway within the hippocampus was significantly augmented both pre-­‐ and post-­‐tetany. However, the effect of tetany was not significant. Despite tetany’s inability to significantly augment the perforant pathway response, the pairing of the septal and perforant pathway stimulations did result in a facilitation of the perforant pathway. This project is funded by NSF grant No. IOS-­‐1048556 and a grant from the Davidson College Research Initiative. Email: [email protected] 59 Sophie Claudel Julio J. Ramirez Analysis of spatial working memory following bilateral hippocampal lesion in rats S.E. CLAUDEL, J.J. RAMIREZ Davidson College One of the hallmark symptoms of AD is debilitating memory loss, which presents with difficulty recognizing previously familiar surroundings and points to a deficit in spatial working memory. The hippocampus is heavily involved in learning and memory, particularly spatial working memory, which is the ability to retain spatial information in working memory and apply it to a task within a short period of time. Conflicting evidence of dorsal or ventral hippocampal involvement in spatial working memory 41 suggests the need to investigate the effect of a complete hippocampal lesion. In this study, an eight-­‐arm radial maze was employed to assess spatial working memory for 6 weeks following bilateral hippocampal lesions. Spatial working memory was impaired for the first 4 weeks of post-­‐operative testing (p < 0.05). This may have been due to a loss of hippocampal place cells or an inability to synthesize information from entorhinal grid cells, which facilitate the formation of a cognitive map. The lack of a cognitive map may contribute to poor performance in the maze, although lesion cases did show improvement over time, indicating possible compensation by the entorhinal cortex or other extra-­‐hippocampal areas. This project was funded by the Davidson Research Initiative, the National Institutes of Health (Grant #MH 060608) and the National Science Foundation (Grant #IOS-­‐1048556). Email: [email protected] 60 J. Anna Juras Robert L. Rosenberg Decreased sodium channel expression during spinal cord regeneration in lamprey J.A. JURAS, A.A. KURUP, R.Y. LEWIS, R.C. PALMARINI, E.S. RICHARDS, Y.J. RODRIGUEZ, R.L. ROSENBERG Earlham College Around 200,000 people live with spinal cord injury (SCI) in the US, with ~12,000 new cases per year. Decreased quality of life and huge health care costs make SCI a devastating condition. Lampreys are a well-­‐characterized vertebrate model for SCI. Unlike higher vertebrates, lampreys exhibit spinal cord regeneration; 10-­‐12 weeks after complete spinal transection they can swim almost normally. New knowledge on lamprey spinal cord regeneration could help identify mechanisms for improved recovery from SCI in humans. Voltage-­‐gated sodium channels (NaV) allow neurons to create and propagate action potentials but excessive NaV activity after injury could cause hyper-­‐excitability and calcium overload that can kill neurons. Lampreys recovering from SCI are resistant to NaV blockers, suggesting that their expression of NaV is changed during SCI. Preliminary transcriptome analysis suggests a decreased expression of NaV during recovery. Thus, decreased NaV expression may be an important for the survival and regeneration of spinal neurons following SCI. This study assessed NaV expression in normal and transected lamprey spinal cords. We used immunofluorescence microscopy to visualize NaV in spinal cord sections before SCI and during recovery from SCI. We optimized conditions to improve the visualization of fine, filamentous structures that we infer are nerve axons. To quantify expression, we asked an observer, uninformed about the experiment, to count the NaV-­‐labeled axons in the micrographs. There was a significant decrease in NaV-­‐labelled axons after injury that started to recover after 11 weeks. We also measured the time-­‐course of the recovery of swimming ability and the effect of a NaV blocker during recovery. The NaV blocker allowed slightly faster recovery. Thus, our preliminary data provide evidence of decreased expression of voltage-­‐gated sodium channels in regenerating lamprey axons and a functional benefit of this decreased expression. Grant support: Great Lakes Colleges Association New Directions Initiative, Marine Biological Laboratory (MBL) Whitman Center awards, Earlham College Ford/Knight Endowment, Stephenson Fund, and Professional Development Fund Email: [email protected] 61 Juyun Kim Yoland Smith, PhD Stereological analysis of GABAergic interneurons in the striatum of a transgenic Huntington’s disease monkey model J. KIM, R.M. VILLALBA, S. JENKINS, A. CHAN, Y. SMITH Emory University-­‐ Yerkes National Primate Research Center, Furman University 42 Huntington’s disease (HD) is an autosomal dominant trinucleotide repeat neurodegenerative disorder caused by the expansion of CAG repeats in the HTT gene. Recent research has been devoted to the development of a transgenic non-­‐human primate model of HD (Yang et al., 2008, Nature 453:921). It is not certain as to whether or not striatal degeneration in these animals is similar to that seen in HD patients. In order to characterize the brain pathology in these monkeys, and compare it with data obtained from postmortem studies of the HD human brain, we undertook a stereological analysis of neuronal counts in the striatum, i.e. the main site of brain pathology in HD, in two HD transgenic monkeys. In primates (including humans), the striatum is made up of the caudate nucleus, putamen, and nucleus accumbens. In severe HD patients, more than 90% of striatal projection neurons (also called Medium Spiny Neurons) degenerate, while the remaining neurons of the striatum, called interneurons, are selectively spared. To determine if similar changes were found in HD monkeys, we assessed the extent of neuronal degeneration in the striatum of two transgenic HD monkeys that expressed exon 1 of the human HTT gene. In a recent study, our colleagues demonstrated a significant loss of striatal projection neurons in these animals (Chen et al., 2013, SFN abstr 528.01). To complement these observations, the goal of this study was to determine if two main populations of striatal GABAergic interneurons that express parvalbumin (PV) or calretinin (CR) immunoreactivity were also affected in these monkeys. To do so, we used antibodies raised against PV or CR to label these neurons throughout the full extent of the striatum in the two HD monkeys and age-­‐
matched controls. We, then, determined the total number of labeled interneurons for each marker in the caudate nucleus and putamen of each animal using the optical fractional method based on unbiased stereological principles. When compared with control monkeys, our preliminary data suggest that there is an overall increase in the total number of both CR-­‐ and PV-­‐positive interneurons in both the caudate nucleus and the putamen of HD transgenic monkeys. Studies are in progress to further characterize these findings and compare those with other populations of striatal interneurons. In conclusion, our data suggest that some of the neuropathological features of transgenic HD monkeys are comparable to those described in the striatum of HD patients. Therefore, transgenic HD monkeys may become a unique “gold standard” model to study HD pathophysiology and therapeutic development. Email: [email protected] 62.1 Paul Dixon Onarae Rice Will blocking the dopamine D3 receptor decrease the expression of PTSD in rats? C. DIXON, W. LORENZO, J. HAYDEN, O. RICE Furman University Previous studies provide evidence that dopamine D3 receptor antagonists and dopamine D3 receptor partial agonists attenuate the expression of fear conditioning in rats. These findings suggest that a dopamine D3 receptor antagonist may be used to reduce the expression anxiety disorders such as posttraumatic stress disorder (PTSD). The present study observed the effect of the novel dopamine D3 receptor antagonist YQA14 on the expression of PTSD. Using a modified single prolonged stress model (SPS), rats (N=40) were exposed to 3 conditioned stimulus-­‐stressor pairings successively (forced swim, restraint, and inescapable shock). Consistent with time dependent sensitization studies, rats were left in their home cages for 7 days following SPS to allow for the development of PTSD-­‐like manifestations. Rats were then placed in a novel environment and exposed to the conditioned stimulus. Freeze time and mass of fecal pellets were used as a behavioral measure of PTSD-­‐like anxiety in the novel environment. Rats that received YQA14 (6.25, 12.5 mg/kg i.p.) prior to exposure to the conditioned stimulus in the novel environment showed significantly lower freeze times as compared to rats that received the vehicle or a higher dose of the drug (25 mg/kg). These findings suggest that YQA14 reduces the expression of PTSD in rats. Email: [email protected] 43 62.2 Will Laurenzo Onarae Rice An investigation of the effects of single prolonged stress (SPS) on male Sprague Dawley rats' propensity to self-­‐administer cocaine W. W. LAURENZO, C. DIXON, J. ZHANG, J. HAYDEN, O. V. RICE Furman University Rationale Exposure to acute physical and psychological stressors induces long-­‐lasting changes within the brain that may increase the risk of psychomotor stimulant abuse and addiction. Literature suggests a high rate of comorbidity between PTSD victims and psychostimulant dependence. Objective The present experiment is designed to determine whether pre-­‐exposure to PTSD-­‐like stressors predisposes rats to cocaine seeking and addiction. Materials and Methods Male Sprague Dawley rats (450-­‐550g) will be randomly assigned to two groups: one control group that receives no stressor, and an experimental group that is subjected to a modified version of the single prolonged stress (SPS) procedure. This paradigm is created to mimic PTSD-­‐inducing stressors, and consists of a 20 minute forced swim test, followed by a two-­‐hour full body restraint period, followed by a 20 minute period of intermittent foot shocks (.5 mA). Four to six weeks after exposure to SPS, both the control and experimental groups will be implanted with chronic indwelling jugular catheters and allowed free access to self-­‐administer cocaine (.75 mg/kg/inf) on a FR1 schedule of reinforcement. Once rats have received 30 infusions per three-­‐hour session for three consecutive days, they are moved to a FR5 schedule of reinforcement, and the cocaine dosage is lowered to .25 mg/kg/inf. After rats display steady bar pressing behavior for three consecutive days under the FR5 schedule, they are moved to a progressive ratio (PR) schedule of reinforcement. Infusions, active lever presses, and inactive lever presses are recorded for five days of three-­‐hour PR sessions. PR breakpoints will be analyzed and compared between groups. Email: [email protected] 63 Megan Willner Renee Chosed Elucidating the role of deubiquitinating enzymes in a yeast model for Parkinson’s disease M.A.H. WILLNER, R.J. CHOSED Furman University Parkinson’s disease (PD) is a neurological disorder that affects the motor neurons within the brain, causing tremors, muscle spasms, and loss of coordination. A key feature of this disorder is an accumulation of the protein alpha-­‐synuclein in the neurons, which eventually causes the neurons to be non-­‐functional due to alpha-­‐synuclein aggregation in the cytoplasm. Yeast share approximately 60% sequencing similarity with human DNA, making them an ideal model organism to study neurological disorders. Alpha-­‐synuclein, tagged with green-­‐fluorescent protein for visualization, was previously integrated into the genome of a strain of Saccharomyces cerevisiae. Previous studies have shown that ubiquitin attaches to excess alpha-­‐synuclein protein which targets alpha-­‐synuclein for proteasomal degradation. In PD, we hypothesize that the ubiquitination of alpha-­‐synuclein may be regulated by deubiquitinating enzymes present in the cell. The deubiquitinating activity of these enzymes would allow an excess of alpha-­‐synuclein to accumulate in the cell, which may yield large protein aggregates. Initial biochemical studies with the PD model yeast treated with a global deubiquitinating enzyme inhibitor support our hypothesis. To determine which specific deubiquitinating enzymes are responsible for the deubiquitination of alpha-­‐synuclein, the genes UBP1, UBP5, UBP6 and YUH1, which encode deubiquitinating enzymes, were individually deleted from the yeast genome. These yeast deletion strains were then examined under a fluorescent microscope to assess aggregation of alpha-­‐synuclein. If protein aggregates are not present, this could be due to the deletion of the deubiquitinating enzymes, which allows the excess alpha-­‐synuclein to be degraded. After the deletion of UBP1, UBP5 , UBP6 and YUH1, alpha-­‐synuclein inclusions were minimal or only localized at the membrane. Further studies with 44 expression of catalytically dead versions of these enzymes have yielded similar yeast phenotypes. These results reveal that regulation of deubiquitinating enzymes may play a role in the development of PD, which could potentially aid in characterizing the biological changes that occurs in neurons in patients with PD. Support provided by: SCICU, HHMI, SC-­‐NIH INBRE Email: [email protected] 64 Michelle Dominguez Abigail Kerr Aerobic exercise is as effective as skilled reach training in improving functional recovery following ischemic insult in C57BL/6 mice. M. DOMINGUEZ, T. MUELLER, R. HOLDEN, M. CURTIS, B. WALL, A.L. KERR Illinois Wesleyan University Stroke is a leading cause of serious, long-­‐term disability with upper limb impairment among the most common, chronic deficits reported in survivors. Current rehabilitative strategies employed in humans are often ineffective in restoring pre-­‐stroke behavioral performance. It has been found that focused rehabilitative training of the impaired limb effectively improves functional outcome in rodent models of ischemic stroke. However, the benefits reported with this training require intensive therapy with relatively long sessions implemented daily. Additional rehabilitative strategies that may be easier to implement in other populations (including humans) need to be established to promote better behavioral recovery following injury. Aerobic exercise has been found to have beneficial effects for both the brain and behavior in humans and rodents, with exercise improving learning and memory in aged populations. The current study investigates the role of aerobic exercise in functional recovery following focal ischemic insult in C57BL/6 mice. A total of 40 mice were trained to criterion on a skilled reaching task, the Pasta Matrix Reaching Task (PMRT), prior to receiving a unilateral ischemic stroke. Following four days of recovery, mice were divided evenly into one of four groups: skilled reach training of the impaired limb (on the PMRT), aerobic exercise, skilled reach training with aerobic exercise, or control procedures. Animals in aerobic exercise conditions had free access to running wheels, while control animals received no direct training of any limb. All mice were in their respective conditions for a total of two weeks and received weekly probe trials of their impaired limb. Our results indicate that aerobic exercise is as effective as skilled reach training in improving functional recovery of the impaired limb. The combined therapy of skilled reach training and exercise was no more beneficial than either condition alone. These findings suggest that aerobic exercise may be a feasible rehabilitative strategy for individuals with persistent upper extremity impairment following stroke. Email: [email protected] 65.1 Vinay Srinivasan Karen Parfitt Aβ25-­‐35 modestly reduces hippocampal long term potentiation in vitro V. SRINIVASAN, S.A. CRAWFORD, J.C. NECARSULMER, I. SOLOMON, L.R. BLEICHNER Pomona College Alzheimer's disease (AD), characterized by severe memory loss and progressively worsening neurocognitive deficits, is one of the most pressing concerns for the aging population of the 21st century. At a molecular level, extracellular oligomerization of beta-­‐amyloids (A-­‐Beta), ~40 amino acid peptides produced by beta and gamma secretase cleavage of the Alzheimer's precursor protein, is thought to be responsible for the pathology. However, much is still unknown about the specific mechanisms underlying the peptide's neurotoxic effects within the brain. Previous work has examined hippocampal long-­‐term potentiation (LTP), an accepted molecular basis of learning and memory, to investigate the role of A-­‐Beta in mediating AD symptoms. The results of these studies have suggested a selective, post-­‐synaptic 45 receptor-­‐mediated mechanism of interaction between A-­‐Beta and the modulators of LTP. As the basis for future work demonstrating the efficacy of neuroprotective compounds, our project sought to demonstrate the purportedly clear inhibitory effect of A-­‐Beta (25-­‐35) on theta burst-­‐induced hippocampal LTP by recording excitatory post-­‐synaptic potentials in the CA1 region. We observed only marginal inhibition of LTP in treated slices compared to control, which differs from the more stark differences found in previous LTP work. However, low [Mg2+] in our baths and/or an overly-­‐strong theta burst protocol might account for the absence of significant inhibition of LTP. Supported by the Pomona College Summer Undergraduate Research Program (SURP) fund and the Pomona College Department of Neuroscience. Email: [email protected] 65.2 Julie Necarsulmer Karen Parfitt Morris Water Maze, Radial Arm Water Maze, and Novel Object Recognition Tasks detect spatial learning and memory deficits in mice J.C. NECARSULMER, L.R. BLEICHNER, I. SOLOMON, V. SRINIVASAN, S. CRAWFORD Pomona College Alzheimer's Disease (AD) is characterized by a progressive loss of cognitive function, notably the impairment of spatial learning and memory. The present study examined three tasks commonly used to test hippocampal-­‐dependent spatial learning and memory—the Morris Water Maze (MWM), Radial Arm Water Maze (RAWM) and the Novel Object Recognition (NOR) tasks, as potentially viable assays for future experiments that will utilize an AD transgenic mouse model. Spatial learning and memory requires the glutamate NMDA receptor. We used the NMDA antagonist MK-­‐801 to examine whether performance on these tasks is impaired, as has been shown by previous groups in this and other rodent strains. For each task, MK-­‐801 (0.05 mg/kg; 5mL/kg in 0.9% saline vehicle) or vehicle alone was administered to C57B/6N mice 30 minutes prior to testing. Deficits in spatial learning and memory were demonstrated by all three tasks when comparing MK-­‐801-­‐treated mice to vehicle-­‐treated controls, indicating that the MWM, RAWM, and NOR task are all viable assays for use in future studies. Email: [email protected] 66.1 Samuel Crawford Karen Parfitt Higher concentrations of Mg++ inhibit the formation of long term potentiation in mouse hippocampal area CA1. S. A. CRAWFORD, V. SRINIVASAN, I. SOLOMON, J. C. NECARSULMER, L.R. BLEICHNER Pomona College An important excitatory neurotransmitter in the brain is the simple amino acid, l-­‐glutamate. Some receptors for glutamate, e.g the N-­‐methyl-­‐D-­‐aspartate (NMDA) receptors, are ion channels that are blocked by magnesium ions (Mg++) at normal resting membrane potentials. We used low (0.85 mM) and high (2.8 mM) concentrations of Mg++ to determine the influence of Mg++ on paired pulse facilitation (PPF) and induction of long-­‐term potentiation (LTP) via the NMDA receptor-­‐-­‐mediated pathway. LTP and PPF were measured in the CA1 area of hippocampal slices prepared from C57 Bl 6N mice. The magnitude of LTP was significantly greater in the low Mg++ concentration. PPF was not affected by the Mg++ concentration, suggesting that Mg++ influences LTP via a postsynaptic role. Future studies will involve a concentration of Mg++ similar to that which occurs naturally in cerebrospinal fluid, in an effort to establish appropriate controls when altering concentrations of other LTP–affecting molecules (e.g. Aβ, the oligomer responsible for forming plaques in the brain, which lead to Alzheimer’s related symptoms.) Email: [email protected] 46 66.2 Lauren Kershberg Karl Johnson The characterization of chondroitin sulfate proteoglycans in Drosophila melanogaster using loss of function mutants and RNA interference to remove CS biosynthetic enzymes. L.H. KERSHBERG Pomona College Chondroitin sulfate proteoglycans (CSPGs) are extracellular matrix molecules involved in several neural processes. CSPGs, protein cores with specific glycosaminoglycan (GAG) sugar side chains attached, serve as repulsive guidance cues during neuronal cell migration in early development. Furthermore, CSPGs have been implicated in a loss of brain plasticity forming critical periods and in the permanence of spinal cord injury. CSPGs have never been identified in Drosophila despite evidence of their existence. Drosophila possess genes, that are conserved across species, for the enzymes that synthesize CSPGs. I have removed the enzymes required to synthesize CSPGs using both loss of function mutations as well as through RNA interference and have found axon guidance defects. Quantification and further analysis of the phenotypic defects are in progress. Fluorescein conjugated lectin that binds to GalNAc residues have been used to visualize CSPG localization in dissected Drosophila embryos. Preliminary results suggest that CSPGs exist in Drosophila and play a role in axon guidance at the midline. Email: [email protected] 67.1 Nirav Mathur Andy Kazama Paradoxical functional facilitation of stimulus-­‐reward association learning in macaques with neonatal perirhinal lesions N. MATHUR, A.M. KAZAMA, M.C. ALVARADO, J. BACHEVALIER Furman University Behavioral studies of monkeys, specifically rhesus macaques (Macaca Mulatta), have illustrated that the rhinal cortex (formed from the perirhinal and entorhinal cortices) is involved with numerous cognitive processes such as object identification and object memory. However, research has shown that damage to this area in adult monkeys spares stimulus-­‐reward learning as well as the ability to shift choice selection when the rewarding value of objects has been modified by satiation (Baxter & Murray, 2001; Thornton et al., 1998). These findings suggested that the rhinal cortex is not a critical part of the neural circuitry mediating stimulus-­‐reward association learning and of flexible choice selection in adult monkeys. However it is not known whether the perirhinal cortex (PRh) may be more critical for the development of stimulus-­‐reward association learning and reinforcer devaluation. Thus, we tested adult animals that had received neonatal lesions of the PRh in a reinforcer devaluation task (Kazama & Bachevalier 2013), measuring stimulus-­‐reward association learning and flexible choice selection. We found unexpectedly, that neonatal PRh damage improved stimulus-­‐reward learning as compared to controls, but their ability to flexibly alter choice did not differ from controls. Thus, PRh does not seem to be involved in flexible decision-­‐making at 4 years of age. This study demonstrates for the first time that early damage to the PRh results in paradoxical facilitation of the ability to learn object-­‐reward associations. Given that this facilitation was not observed in animals with adult-­‐onset rhinal lesions, our data suggest that, due to greater brain plasticity in infancy, the neonatal lesions may have promoted greater brain reorganization. Email: [email protected] 47 67.2 Alyssa Althoff David Hollis Effects of Notch, Wnt, and IL-­‐4 modulation on muscle progenitor cell Pax7 expression and tube formation A. ALTHOFF, Z. WANG, S. SOKER Furman University & Wake Forest Institute for Regenerative Medicine Stem cell therapies have shown to be viable options for restoring muscle function in areas of large muscle loss, as experienced in muscle degenerative diseases such as compartment syndrome. Muscle progenitor cells (MPCs) are potential candidates for cell therapies, as they can be isolated from the donor, expanded in culture, and form muscle fibers at the site of muscle damage. During regeneration, MPCs proliferate and differentiate, before fusing with one another, as de novo myotubes, or into existing nascent muscle fibers. Our former work showed first, that MPCs express the muscle stem cell marker Pax7, and, second, that MPCs are able to form myofibers, or to engraft, in vivo. However, the in vivo engraftment was not as efficient as engraftment from freshly isolated muscle stem cells, which yields the most potent engraftment after cell implantation. Thus, one challenge of using MPCs is finding a method to greatly increase the in vivo engraftment efficiency. Through microenvironment modulation, it is possible to control the MPCs fate, thus increasing their in vivo engraftment efficiency. We hypothesized that modulating Notch, Wnt and IL4, the most important signaling pathways governing muscle stem cell fate from cell expansion to myotube fusion, will increase MPC engraftment efficiency. As such, we introduced signaling pathway modulating drugs targeting the Notch, Wnt and IL4 pathways to MPCs during cell proliferation and myotube formation. We examined Pax7 frequency, myotube formation, and cell fusion to evaluate the efficacy of signaling pathway modulating on MPC engraftment ability. To test Pax7 expression, drugs targeting Wnt, Notch, and IL4 signaling pathways were added to cultured MPCs for 4 days, then Pax7 immunofluorecent staining was performed. To test myotube formation, MPCs were allowed to grow to high density to initiate nascent myotube formation. Drugs were then added for 4 days, and images of formed myotubes were obtained and analyzed. To evaluate cell fusion, drugs were added to GFP+ MPCs for 4 days during cell proliferation, then the GFP MPCs were added to Tomato Red+ nascent myotubes. Green-­‐red mixed fluorescent myotubes, resembling the donor-­‐host myofibers formed in vivo, were then quantified. Our data showed that blocking the Wnt pathway (drug XAV939, 3.3 uM) demonstrated trends towards increased Pax 7+ expression in MPCs, and a significant increase in cell fusion ability of MPCs with nascent myotubes, compared to a control (without drug). Conversely, activating the Wnt pathway (Bio, 5 uM) or blocking the Notch pathway (DAPT, 5 uM) decreased Pax7 expression, and had no effect on cell fusion or tube formation. Altering the IL4 pathway (IL4, 10 ng/ml) had no effect on Pax 7+ expression, tube formation or cell fusion. These studies on Pax7 expression and cell fusion can serves as predictors of in vivo cell engraftment efficiency. Out data suggested blockade of Wnt during MPC expansion will potentially increase cell engraftment in vivo. Email: [email protected] 68 Elyse Barnard Mark R. Stefani The neurosteroid pregnenolone sulfate interacts with the NMDA receptor antagonist MK-­‐801 to impair cognitive flexibility and spatial working memory in a rat model of schizophrenia E.D.H. BARNARD, J.K. GERHMANN, D.M. PRIOR, M.R. STEFANI Middlebury College Schizophrenia is a chronic disorder characterized by debilitating cognitive impairments. Most affected are the “executive functions” crucial for the temporal organization of goal-­‐directed behaviors. These cognitive abilities, which include including working memory and cognitive flexibility, require brain networks sharing a common node in the prefrontal cortex (PFC). Recent research has linked these cognitive deficits to reduced PFC inhibitory signaling by the neurotransmitter GABA, leading to the hypothesis that reduced 48 inhibitory regulation of glutamatergic principle neurons results in disorganized information processing and cognitive impairments. We used male Sprague-­‐Dawley rats to examine the cognitive effects of pregnenolone sulfate, a neurosteroid that modulates GABAA receptors to reduce GABA-­‐mediated inhibition, and MK-­‐801, an NMDA glutamate receptor antagonist commonly used to induce schizophrenia-­‐
like cognitive deficits in rodents. MK-­‐801 is believed to act, in part, through inhibitory actions on GABA neurons. Rats were tested for extradimensional (ED) set-­‐shifting ability, a measure of cognitive flexibility, or spontaneous alternation behavior (SAB), a measure of spatial working memory. Rats received bilateral intra-­‐medial PFC injections (0.5 uL/hemisphere) 20 min prior to the ED shift phase of the set-­‐shift task or the single SAB session. Injections consisted of a vehicle solution, MK-­‐801 (0.3 or 3 µg/hemisphere), or pregnenolone sulfate (0.10, 1.0 or 10 ng/hemisphere), alone or in select combinations. We observed that MK-­‐801 alone at a dose of 3.0 ug but not 0.3 ug impaired cognitive performance relative to vehicle-­‐
injected controls, in the set-­‐shift task by increasing the trials required to reach a learning criterion and increasing the amount of perseverative responding to the first discrimination rule learned, and in the SAB task by decreasing the percent alternation score. Pregnenolone sulfate also dose-­‐dependently impaired task performance. In the set-­‐shift task, the highest dose of pregnenolone sulfate produced significant impairments; in the SAB task, the intermediate dose significantly impaired. Co-­‐administration of individually ineffective doses of MK-­‐801 and pregnenolone sulfate impaired task performance relative to vehicle-­‐injected controls in the set-­‐shift task. This pattern of results, in which two neuromodulatory compounds know to act by decreasing GABA-­‐mediated inhibitory tone interact to produce deficits in behavior task dependent on prefrontal function, provide further support for the hypothesis that aberrant GABAergic signaling underlies schizophrenia-­‐associated cognitive deficits. Email: [email protected] 69 Adam Ghoweri Cecilia M. Fox The neuroprotective potential of intranasal DNSP-­‐11 on nigral dopamine neurons against intrastriatal 6-­‐hydroxydopamine neurotoxicity A. GHOWERI, C.M. FOX Moravian College Using both in vitro and in vivo studies, glial cell line-­‐derived neurotrophic factor (GDNF) became known as one of the more promising neurotrophic factors in its ability to protect dopamine neurons against neurotoxic insult in animal models of Parkinson’s disease (PD). The proprotein version of GDNF has been post-­‐translationally processed into a dopamine neuron stimulating peptide, known as DNSP-­‐11. DNSP-­‐11 has been shown to be neuroprotective against TaClo, MPP+ and an intranigral 6-­‐hydroxydopamine (6-­‐
OHDA) lesion in rat models of PD. This research project used a different approach to introducing DNSP-­‐11 into the animal post-­‐lesion. An intranasal DNSP-­‐11 technique was used to assess the protection of nigral dopamine neurons against the more progressive intrastriatal lesion of 6-­‐OHDA. Twenty Fisher 344 rats were divided into the following groups: citrate + 6-­‐OHDA and DNSP-­‐11 + 6-­‐OHDA. Citrate or DNSP-­‐
11was delivered intranasally for eight weeks post-­‐lesion. The foot fault and cylinder tests were performed to assess behavior improvements following treatment. Brain tissue will be processed for tyrosine hydroxylase immunocytochemistry and dopamine cell survival will be quantified via stereology. Email: [email protected] 70 Alec Hyde Deborah S Kreiss Neonatal exposure to clomipramine is a behavioral rat model of Obsessive Compulsive Disorder offering not only face validity, but also predictive validity for the GABA agonist diazepam, the norepinephrine uptake inhibitor desipramine, the serotonin uptake inhibitor fluoxetine, and the serotonin 5HT2 antagonist mianserin 49 A.L. HYDE, L.J. KASPARSON, L.S. LAIKS, K.M. CRAIG, S.E. FRANK, H.S. MULDER, D.S. KREISS Colgate University, Psychology Dept, Neuroscience Program Obsessive Compulsive Disorder (OCD) -­‐ a psychiatric disorder affecting 2-­‐3% of the population -­‐ is characterized by persistent anxiety accompanied by overwhelming urges to perform repetitive behaviors. Current pharmacological treatments for OCD are only effective in 40-­‐60% of patients and have an 8-­‐10 week delayed onset. Animal models with both face and predictive validity are imperative in the field of OCD so that better treatment avenues can be developed. The current study explores a novel rodent model of OCD. Male rats were either administered clomipramine (15 mg/kg x 2, “neoClom”) or saline (1 ml/kg X 2, “neoSaline”) during neonatal Days 9-­‐16. Head poking in the hole-­‐board as well as marble burying and a novel checking behavior were assessed at Day 80-­‐82 and Day 208-­‐210, respectively. NeoCloms (n=14) buried significantly more marbles than neoSalines, but did not differ in other observed behaviors. In our 2013 studies, neoCloms (n=26, assessed at Day 70-­‐72) significantly buried more marbles, checked more marbles, and exhibited more repetitive head pokes. Our results suggest that younger behavioral testing and a larger group size facilitate observation of more robust differences. In 2013, our lab demonstrated that 14-­‐day repeated fluoxetine (10 mg/kg) normalized hole-­‐board and marble behaviors of neoCloms, whereas repeated desipramine (5 mg/kg) or saline did not, mirroring the clinical efficacy of these agents in OCD. In the current study, we show that acute injection of the GABA agonist diazepam (1, 8 mg/kg) did not alter behaviors of neoClom rats. However in the neoSalines, head dipping in the hole-­‐board was significantly increased by 1 mg/kg diazepam and decreased by the 8 mg/kg dose. Our results parallel the ineffectiveness of diazepam at alleviating symptoms of OCD patients and the typical behavioral alterations induced by GABA agonists in humans w/o OCD. Additional studies showed that the effect of the serotonin receptor antagonist mianserin on our rats is analogous to the drug’s clinical effectiveness. In the neoCloms, mianserin significantly decreased the number of head pokes, holes, and repetitive head pokes – whereas in the neoSalines, mianserin had no effect. Acute injection of the serotonin agonist mCPP (0.4, 0.8 mg/kg) significantly decreased marbles checked in both neonatal groups, but did not alter other behaviors. These results are incongruent with the ability of mCPP to exacerbate symptoms in OCD patients and warrant further exploration. These data confirm the face validity of the neoClom model of OCD and provide further demonstration of the predictive validity of this unique permanent, multi-­‐
symptom behavioral model of OCD. Source of Support: Colgate University: Neuroscience Program and Research Council www.colgate.edu/deb-­‐kreiss Email: [email protected] 71.1 Camille Pham-­‐Lake Jennifer Larimore Experimental techniques to evaluate the neurotoxicity of environmental chemicals C. PHAM-­‐LAKE, J. BRADNER, W. CAUDLE Agnes Scott College There are over 80,000 chemical contaminants in the environment, most in the form of pesticides and industrial solvents. Many are known to contribute to the development of neurological diseases. Research is essential for understanding their relationship to disease and the neural functions they disrupt. This study focused on polychlorinated biphenyls (PCBs). Due to their physiochemical characteristics they persist in the environment and are significant contributors to deficits in both cognitive and motor functions. Exposure to PCBs has a high correlation with damage to the dopamine system and incidences of Parkinson’s disease. Given the conformational diversity of the PCB’s this investigation was structured to further explore their differing neurotoxicity’s through toxicity assays on both the SK-­‐N-­‐SH cell line and primary cultures, immunohistochemistry, and immunofluorescenceto investigate the neurotoxicity of PCB 52 and Aroclor 1254 on mice tissue. We hypothesize that PCB 52 and Aroclor 1254 will show differing levels of toxicity and differing levels of toxicity on specific brain regions. Early findings suggest a divergence in both the toxicity of PCB 52 and Aroclor 1254 and neural structural differences in cortex and midbrain tissues. This experimental platform can be used to further assess other environmental toxicants. 50 71.2 Khallyl Oliver Jennifer Larimore Temporal pattern of GABAA receptor delta and gamma-­‐2 subunit expression in the suprachiasmatic nucleus of male Syrian hamsters K. OLIVER, J. WALTON, E. ALBERS Georgia State University The suprachiasmatic nucleus (SCN) is the region of the hypothalamus, situated just above the optic chiasm, responsible for controlling the circadian rhythms of the body. It is widely known that gamma-­‐
amino butyric acid (GABA) modulates SCN activities, while the SCN mediates circadian rhythms via entrainment. GABAA delta and GABAA gamma-­‐2 subunits co-­‐regulate each other in the SCN. We hypothesize that the extrasynaptic GABAA delta receptor is more abundant during the dark phase as opposed to GABAA gamma-­‐2, which should be active in light, based on the knowledge that GABAA delta is more sensitive than GABAA gamma-­‐2 and is able to down regulate gamma-­‐2. Adult male Syrian hamsters were ordered from Charles Rivers Laboratories. Hamsters were singly-­‐housed and entrained to a 14:10 light/dark (L: D) cycle over a course of 2-­‐4 weeks. For constant dark (D: D) conditions, hamsters were singly-­‐housed and held in a 14:10 light cycle for two weeks, then placed in a constant dark setting for 9-­‐14 days. Hamsters were also placed in constant light conditions (L: L) for 9-­‐14 days as well. Activity was measured via wheel running. Hamster suprachiasmatic nuclei were collected via micropunch. RNA was extracted with Trizol, reversed transcribed into cDNA library, then expression of GABAA delta and gamma-­‐2 was assessed using a gene specific primer for RT-­‐PCR. Relative gene expression levels for each gene of interest were normalized to 18s expression. Relative mRNA results showed opposite of hypothesis, with GABAA gamma-­‐2 being more readily expressed in light hours and GABAA delta being more expressed in dark hours. 72 Gifty Dominah Gunnar Kwakye Acute exposure to Chlorpyrifos generates oxidative stress and mitochondrial dysfunction in a striatal cell model of Huntington’s disease. G.A. DOMINAH, G.F. KWAKYE Oberlin College In spite of the genetic cause of Huntington’s disease (HD), emerging evidence strongly suggests environmental influence on the age of onset, progression, and severity of the disease. However, the identity of the environmental risk factor is currently unknown. Recognizing the similarities in the pathophysiological mechanisms between HD and pesticide neurotoxicity, we hypothesized that the common agricultural pesticide chlorpyrifos (CPF) would exhibit disease-­‐toxicant interaction and reveal the influence of CPF in HD neuropathophysiology. We investigated the effects of acute CPF toxicity and its principal metabolites chlorpyrifos oxon (CPO) and 3,5,6-­‐trichloropyridinol (TCP) in an established murine striatal STHdh cell model of HD by assessing cell viability, reactive species production, mitochondrial membrane potential, antioxidant buffering capacity, and energy homeostasis, as well as antioxidant mediated neuroprotection. Following a 48 hours exposure to the metabolites CPO and TCP, we observed no significant dose and genotypic differences in cell survival. Interestingly, expression of mutant HD resulted in increased dose-­‐dependent susceptibility to CPF exposure and production of reactive species compared to wild-­‐type cells. Furthermore, we report that the mutant HD induced vulnerability to CPF exposure is mediated through diminished antioxidant buffering capacity, enhanced production of free radicals, decrease mitochondrial function, and energy production. To further investigate the possible neuroprotection of HD cells against CPF neurotoxicity, we treated STHdh cells with N-­‐acetylcysteine (NAC) and observed that the CPF induced toxicity was significantly ameliorated in mutant HD cells. These 51 results strongly suggest that mutant HD and CPF exhibit a disease-­‐toxicant interaction to cause enhanced striatal neurotoxicity via oxidative stress and mitochondrial dysfunction that could exacerbate the neurodegenerative processes in HD. Email: [email protected] 73 Brandon Schurter Christian Fink Synchronization properties of heterogeneous neuronal networks B. SCHURTER, M. ZOCHOWSKI, V. BOOTH, C.G. FINK Ohio Wesleyan University Highly connected neurons, called hub cells, are thought to contribute to certain forms of epilepsy and have also been shown to orchestrate synchrony in the hippocampus of developing rats. How hub cells are capable of hijacking networks to synchrony is not well understood. We hypothesize that the excitability type of hub cells may be an important factor. In general, neuronal excitability (which characterizes how neurons respond to input) falls into two categories, Type I and Type II, with networks of only Type II neurons synchronizing very well, and networks of only Type I neurons synchronizing rather poorly. We used computer simulations to investigate the synchronization properties of networks with a mixture of Type I and Type II neurons. We were particularly interested in the effect of placing Type II neurons as hub cells in the network. The results of these simulations show that relatively few Type II neurons are capable of hijacking the network to synchrony when they are placed as hub cells, but not otherwise, indicating that Type II cells could play a role in generating epileptic seizures. Email: [email protected] 74.1 Jenny Moncion Mimi L. Phan Cholesterol-­‐lowering drugs alter neuronal morphology in a juvenile songbird system J. MONCION, S.C. TSOI, A. QURESHI, C.L. PYTTE, D.S. VICARIO, M.L. PHAN Queens College CUNY; Rutgers University; Graduate Center CUNY; Hostos Community College CUNY Statins are widely-­‐prescribed drugs that lower cholesterol by inhibiting the rate-­‐limiting enzyme in cholesterol synthesis. Recently, there has been an increase in obesity and high cholesterol among children and adolescents. In response, the FDA has approved the use of 4 statin drugs (lovastatin, simvastatin, pravastatin, and atorvastatin) for children as young as 8 years old who have a genetic risk of high cholesterol. Although statins block cholesterol synthesis primarily in the liver, the brain also produces an independent source of cholesterol and statins have been shown to cross the blood brain barrier, and alter brain cholesterol. This is a potential concern given that some adult statin users have reported problems with memory, concentration, and learning. While most studies have focused on neuroprotective properties of statins in injured or diseased brains, effects of statins in healthy brains are inconclusive. Given the cognitive complaints of some adults, we speculated that statins may have an even greater effect on the developing brain of juveniles and developed a songbird model to test this idea. Earlier work from our laboratories showed that juvenile zebra finches treated with atorvastatin (Lipitor®) had impaired song learning and memory. Here, we are investigating whether atorvastatin affects the development of young neurons in the nucleus HVC, a brain region necessary for song learning, which continues to incorporate new neurons throughout life. To assess effects of atorvastatin on new neurons, one group of birds (n=6) was given a daily dosage of 40 mg/kg of Lipitor® in water while age-­‐matched control birds (n=7) received the same volume of water. Birds were treated for 65 days during the critical period for song learning. Brains were then processed using immunohistochemistry to label cells that expressed doublecortin (DCX), a microtubule protein present in young neurons for 20 days, we used Neurolucida software (Microbrightfield) to trace the somas of DCX-­‐expressing neurons in HVC. We found that DCX-­‐
expressing neurons in statin-­‐treated birds were flatter, more convoluted, and had rough or pocked cell 52 membrane. These findings suggest that statins may alter the structure of newly formed neurons, or the rate of development of young neurons, perhaps by interfering with cholesterol availability for membrane formation. We propose that altered neuron structure may be one factor contributing to the song learning and memory deficits we observed during juvenile statin treatment. Email: [email protected] 74.2 Alicia Barrientos Mimi L. Phan Cholesterol-­‐lowering drugs alter neuron migration in a juvenile songbird model system. A.C. BARRIENTOS, S.C. TSOI, J. MONCION, A. QURESHI, S. RIBEIRO, C.L. PYTTE, D.S. VICARIO, M.L. PHAN Queens College, CUNY; The Graduate Center, CUNY, Rutgers University Statins are a class of drugs that inhibit cholesterol synthesis. In 2007, the Food and Drug Administration approved 4 statins for pediatric populations at risk for high cholesterol. Among these statins is atorvastatin (Lipitor®), which crosses the blood brain barrier in moderate levels, potentially impacting brain cholesterol. Diminished membrane cholesterol in the brain has been found to affect synaptic vesicle exocytosis, neuronal cytoarchitecture and synapse formation. However, the effects of long-­‐term statin use on cognition and neural substrates in juveniles have not been thoroughly evaluated. In vitro studies testing the effects of statins on brain tissue have provided conflicting results, including decreasing and also increasing new neuron proliferation and survival. Here we used a songbird paradigm to study the effects of long-­‐term statin use in juveniles on learning and neurogenesis in vivo. Zebra finches learn their vocalizations by imitating a tutor, and continually produce new neurons that are incorporated throughout the telencephalon. Earlier work from our labs found deficits in sensorimotor learning in statin-­‐treated juveniles. To determine whether altered neurogenesis may be a mechanism contributing to these learning impairments, we examined new neurons in the song motor pathway, focusing on new neuron morphology, location, and distances from the generative region of the lateral ventricle. Birds were given oral doses of 40 mg/kg of Lipitor® in water (n=6) and control birds given the same volume of water (n=7) throughout the critical period for song learning (daily, for 65 days, bird ages 35-­‐100 days). We labeled mitotically active cells with bromodeoxyuridine (BrdU) and used immunohistochemistry to visualize BrdU, the neuron-­‐specific protein Hu, and also doublecortin, a marker of young neurons. We found that neuronal phenotypes differed between statin-­‐treated and control birds and that phenotypic measures were associated with neuronal location relative to the ventricle. New neurons overall were also significantly further from the lateral ventricle than new glia (BrdU+Hu-­‐ cells) in controls, but not in statin-­‐treated birds. We suggest that Lipitor® may be affecting neural cytoarchitecture as well as the migration and incorporation sites of new neurons, perhaps as a consequence of lowered brain cholesterol. Email: [email protected] 75 Rachel Laufmann Jill Weimer Evaluating the therapeutic benefits of CRMP2-­‐modulating compounds in the treatment of lysosomal storage disorders R.N.LAUFMANN, H. MAGEE, D. TIMM, S. BAACK, J.M.WEIMER Sanford Research Neuronal ceroid lipofuscinosis (NCLs) are a family of lysosomal storage disorders characterized by visual, motor and cognitive decline, behavioral problems and premature death. Pathologically, NCLs result in accumulation of autofluorescent material in the lysosome, neurodegeneration, disruptions in lysosomal enzyme function, and robust glial activation. The variant late infantile form of NCL, vLINCL, results from mutations in the gene CLN6. Our lab has recently demonstrated that the CLN6 protein functions as a part 53 of a novel complex (called the CCK complex) which also contain CRMP2 and kinesin light chain 4 (KLC4). CRMP2 functions to specify axon/dendrite fate and regulates cargo transport through an array of additional binding proteins. Disruption in CRMP2 function has been implicated in a host of neurological disorders, including the NCLs. In this study, we utilize a naturally occurring mouse model of vLINCL (the Cln6nclf mouse) to screen potential therapies that act by modulation CRMP2 stability and function. The lanthionine ketimine derivative lanthionine ketminine ethyl ester (LKE) is a cell permeable, brain penetrating metabolite that specifically targets CRMP2, it stabilizes it’s interactions with a host of binding partners. We will present evidence that indicates LKE treatment starting at weaning (21 days) improves visual acuity, motor function and survival, while decreasing glial activation. Additionally, we have set about performing a comprehensive pathological examination of non-­‐CNS tissues to determine whether there are organs affected by this rare disease aside from the brain. It is our hope that these preliminary studies will provide essential data to guide future compound selection and could launch a new drug development program for NCL. 76 Nate Vinzant Gina Forster Nanotechnology to deliver neuropeptides to the brain: Distribution and effects on anxiety-­‐like behaviors N. VINZANT, J. SCHOLL, C-­‐M. WU, T. KINDLE, V. KUMARASWAMI, R. KOODALI, G. FORSTER Sanford School of Medicine, Division of Basic Biomedical Sciences The corticotropin-­‐releasing factor type 2 receptor (CRF2) represents a novel target for treating anxiety states. Direct infusion of CRF2 receptor antagonist antisauvagine (ASV) into the brain immediately reduces anxiety states in rodent models of early life stress and drug withdrawal. However, like many neuropeptides, ASV cannot cross the blood brain barrier (BBB) if administered systemically. Nanoparticles such as iron oxide can cross the BBB and can be designed to carry drug cargo. This study examined whether iron oxide nanoparticles with ASV cargo cross the BBB by determining the distribution within the rat brain following systemic administration. Iron oxide nanoparticles (Fe2O3) were synthetized, size confirmed by transmission electron microscopy (5.0 +/-­‐ 1.1 nm), and were functionalized with 3-­‐aminopropyltriethoxysilane (1:4 Fe2O3:APTES). Next, ASV (10:1 Fe2O3:ASV) was attached to the functionalized iron oxide nanoparticles and monitored hourly for five hours with fourier transform infrared (FTIR) spectroscopy to ensure stability of the nanoparticle+ASV complex over this time frame. Nanoparticle solution (87.7 μg /kg Fe2O3) with FITC tag, with or without ASV (200 μg/kg) was injected (ip.) 30 minutes prior to transcardial perfusion and brain fixation. Sections throughout the brain were processed using immunohistochemistry and imaged with confocal microscopy. Sections were analyzed for nanoparticle association with neurons (NeuN), with CRF2 receptors, and with iron-­‐related proteins such as ferritin and transferrin. Results suggest systemically-­‐administered nanoparticle with ASV associates with neurons, including those that express CRF2 receptors. To determine the anxiolytic effect of ASV delivered as nanoparticle cargo, rats were pretreated with either amphetamine (2.5 mg/kg) or saline and underwent two weeks withdrawal, which has been shown to increase anxiety-­‐like behaviors. Rats were either treated with nanoparticle+ASV (87.7 μg /kg Fe2O3; 200 μg/kg ASV, ip.) or nanoparticle+vehicle 30 mins prior to testing on the elevated plus maze (EPM). Results thus far suggest that ASV delivered by nanoparticles reduces anxiety-­‐like behaviors. Overall, the findings demonstrate a novel approach to drug delivery across the BBB and provide insight as to the neural distribution and efficacy of drug treatments delivered via nanotechnology. Support: USD Nelson Faculty Research Grant to GF and RK; NIH RO1 DA019921 to GF; USD UDiscover undergraduate research fellowship to NV. 54 77.1 Zahra Melendez Steven Threlkeld Effects of early working memory experience on neuronal morphology within the prefrontal cortex of rats with neonatal hypoxia-­‐ischemia Z.M. MELENDEZ, K.M. MELENDEZ, C.M. GAUDET, S.W. THRELKELD Rhode Island College Hypoxia-­‐ischemia (HI) is low oxygenation to the brain paired with low blood supply that can disrupt normal patterns of brain development. HI injury is characterized by many long-­‐term cognitive and behavioral deficits including working memory. Neuronal plasticity do to early sensory or learning experience has been suggested to facilitate recovery of function after neonatal brain injury. Plasticity is the ability for the nerves system, more specifically neurons, and their synapses to modify their function and morphology due to experiences, which in turn correlate with changes in behavior. The objective of the present study was to investigate the effects of neonatal hypoxia-­‐ischemia on the morphology of layer five pyramidal neurons within the prefrontal cortex (Cg3) of rats with or without early life working memory experience (postnatal day 36-­‐61). We hypothesized that both HI and sham subjects exposed to 20 days of working memory training, using an 8-­‐arm radial water maze, early in life would show distinct morphological changes in Cg3 pyramidal neurons. Findings suggest that early life working memory training regulates shifts in neuronal morphology following neonatal brain injury. This work was funded by the Eunice Kennedy Shriver National Institute of Child Health & Human Development of the National Institutes of Health under grant #R15HD077544. 77.2 Keyshla Melendez Steven Threlkeld Assessment of neuronal morphology in the striatum of rats following neonatal hypoxic-­‐
ischemic injury and early life working memory experience K.M. MELENDEZ, C.M. GUADET, Z.M. MELENDEZ, S.W. THRELKELD Rhode Island College Low blood oxygenation and reduced flow (hypoxia-­‐ischemia (HI) respectively) to the developing brain leads to long-­‐term behavioral deficits including impairments in working memory. Currently, limited treatment options are available for infants at risk for HI injury. However, research suggests that early life experience (e.g., complex housing, auditory stimulation) presented during critical developmental windows can lead to the reorganization of some systems in the brain improving recovery of function following neonatal brain injury in rats. The current study sought to investigate the influence of early life (Postlatal day (P) 36-­‐61) working memory experience on plasticity of medium spiny neurons in the striatum. Effects of HI injury on medium spiny projection neurons of the striatum have not been studied in depth and little information in known regarding the effects HI injury on this central brain structure. Based on work suggesting the involvement of a cortical-­‐striatal-­‐hippocampal circuit in working memory, we hypothesized that early life working memory experience would result in morphological changes of medium spiny neurons in rats with and without neonatal brain injury. Medium Spiny neuron morphology was assessed and dendritic length and other features denoting branch complexity were analyzed. Results suggest that the striatum is affected by neonatal HI injury and that early experience may lead to morphological changes in medium spiny neurons. This research was supported by the Eunice Kennedy Shriver National Institute of Child Health & Human Development of the National Institutes of Health under grant #R15HD077544. 55 78 Sana Sajjad Bruce Gray Free radicals and cell survival in models of neurodegenerative disease S. SAJJAD, C.A. COUGHLIN Simmons College Beta-­‐amyloid is a protein associated with both Alzheimer’s disease and glaucoma as the protein forms insoluble oligomers and plaques in nervous and retinal tissue, leading to cell death. Previous work in the Gray lab has supported the hypothesis that beta-­‐amyloid also has a non-­‐pathological function as it plays a role in modulating cholinergic transmission. Beta-­‐amyloid (Ab) functions as a neuromodulator through a second messenger pathway involving nitric oxide (NO), cGMP and the activation of PKG. To investigate the connection between the pathological role of beta-­‐amyloid leading to cell death and this pathway, embryonic avian retinal cultures were analyzed for survival over time in culture using a neutral red assay. Control primary cultures of 14 day chick retinal cells did not significantly differ from those with added NO (co-­‐incubation with a nitric oxide donor) unless 1uM diethyl,dithiocarbamate (DDC) an inhibitor of superoxide dismutase (SOD) was added. SOD is an enzyme that quenches free radicals. This implies that any free radical formation by NO or Ab is normally rendered harmless (over this time period) by SOD in healthy cells. The decreased survival of DDC and NOD-­‐treated retinal cells was reversed by co-­‐incubation with 5-­‐amino salicylate (5-­‐AS), a specific inhibitor of peroxynitrite formation. Previous work has shown that 5-­‐AS does not reverse effects of Beta Amyloid or NODs on transmitter release. This implies that peroxynitrite formation may factor only in a pathway leading to pathology and cell death. Supported by Simmons SURE (Simmons Undergraduate Research Experience) program to CC, SS and a SURPASs (Summer Undergraduate Research Program At Simmons) to SS. Email: [email protected] 79 Subrina Bisnauth David Ruskin Ketogenic diets improve behavior in the EL mouse model of autism S. BISNAUTH, S.L. SLADE Trinity College The ketogenic diet (KD) is a high fat, very low carbohydrate metabolic therapy that has been shown to significantly reduce seizures in persons with epilepsy which is often comorbid with autism. We previously showed that a strict KD improves symptoms in the BTBR mouse model of autism. KD feeding has been showed to reduce seizures in the EL mouse model of epilepsy co-­‐morbid with autism. Here we examine KD feeding effects on autistic symptoms in EL mice. EL mice of both sexes were fed moderate or strict KDs (or control chow) ad libitum from 5 weeks of age. At 8 weeks of age, behavioral testing began. EL mice on control chow were not significantly social in the 3-­‐chamber test; however, female mice fed either KD became significantly social. Female EL mice on control chow had abnormally high levels of self-­‐directed repetitive behavior (grooming), which was lowered by both KDs. No behaviors of male EL mice were improved by KD feeding. The strict KD reduced growth and blood glucose and elevated blood ketones in females, while only elevating blood ketones in males. The mild KD increased body weight and blood ketones, but did not change blood glucose in females. KD-­‐related behavioral improvements are therefore not dependent on caloric restriction or on lowered blood glucose; elevated blood ketones might be necessary but not sufficient. These data add to the growing literature that KDs could be a valuable treatment for humans with autism. Email: [email protected] 56 80.1 Nick Toker Sara Lagalwar Propagation of mutant ataxin-­‐1 protein in an SCA-­‐1 cell culture model N. TOKER, M. MOOG, S. LAGALWAR Skidmore College Spinocerebellar Ataxia Type-­‐1 (SCA1) is a neurodegenerative disease characterized by loss of Purkinje neurons of the cerebellum, and produces a variety of symptoms including the adoption of an ataxic gait. Affected cells accumulate aggregates of the protein Ataxin-­‐1. These aggregates remain localized intracellularly, although evidence in other neurodegenerative disease fields is beginning to emerge that protein aggregates can propagate from cell to cell. Preliminary experiments using a cerebellar cell line transfected with RFP-­‐tagged mutant Ataxin-­‐1 support this conclusion, indicating that Ataxin-­‐1 aggregates are capable of leaving the nucleus and potentially translocating the plasma membrane. Future experiments will further characterize the mechanism behind the aggregate propagation and determine the role of protein aggregate propagation in disease pathogenesis. Email: [email protected] 80.2 Austin Ferro Sara Lagalwar Mitochondrial OXPHOS dysfunction and treatment in spinocerebellar Ataxia Type 1 A. FERRO, J. ZHANG, E. CARBONE, S. LAGALWAR Skidmore College Spinocerebellar Ataxia Type 1 (SCA1) is a debilitating neurodegenerative disease caused by an abnormally expanded polyglutamine tract (PolyQ) within the ataxin-­‐1 gene. While extensive work over the last two decades has illuminated many of the mechanisms involved in disease progression, treatment remains elusive. We posit that mitochondrial deficits may exacerbate disease progression, and therefore treatment aimed at alleviating these deficits may have potential to slow disease progress. Multiple neurodegenerative disorders including Alzheimer's, Parkinson's and Huntington's disease have been linked to compromised oxidative phosphorylation (OXPHOS) function within complex I of the mitochondrial electron transport chain, increasing reactive oxygen species (ROS) which has long been associated with neurodegenerative disorders. Due to SCA1’s selective neurodegeneration of high energy demand Purkinje cells and the correlation of neurodegeneration with complex I dysfunction, the mitochondria represent potential therapeutic targets. Our experiment aims to bypass complex I in a SCA1 transgenic mouse model using a complex II (succinate ubiquinone oxidoreductase) electron donor, succinic acid (SA). Through chronic treatment with SA, we expect to see improved behavior and pathology compared to non-­‐treated mice. This expected attenuation of disease progression will be assessed through motor and balance behavioral paradigms such as the rotarod and footprint analysis, and immunofluorescent analysis of Purkinje dendritic length and Purkinje mitochondrial distribution. This study intends to exhibit mitochondria as a therapeutic target for neurodegenerative disease as means of slowing disease progression and therefore improving the quality of life for persons with neurodegenerative diseases. Email: [email protected] 81 Kelly Hewes Kimberley A. Phillips Exploring cognitive dysfunction in the marmoset EAE model K. HEWES, K.A. PHILLIPS Trinity University 57 We conducted a pilot study to investigate the effect of exercise on cognitive dysfunction in an animal model of multiple sclerosis (MS). Twelve common marmosets (Callithrix jacchus) were randomly assigned such that eight were injected with MOG34-­‐56 to induce EAE and four were injected with a placebo. Observational health assessments were taken of subjects daily to monitor for symptom progression and euthanasia criteria. Maximum duration of experimentation for each subject was 10 weeks. Of the subjects, six (two control and four MOG) received 30 minutes of moderate aerobic exercise three times weekly via a monitored treadmill. To assess changes in working memory, subjects were also tested on a radial arm maze once weekly during the duration of the experiment. Biological changes will be analyzed to assess brain derived neurotropic factor and glial fibrillary acidic protein using blood serum samples collected at several timepoints during the experiment. Brain tissue will be harvested; one hemisphere will be used for histological analysis and the other hemisphere frozen at -­‐80° C for conducting Western Blots. Additionally, spleen and lymph tissues will be collected for immunological analysis. 82 Natalie Fettinger James R. Moyer Effect of intrahippocampal infusion of Apoaequorin on cytokine protein expression N.B. FETTINGER, S.C. MICHELS, E. L. ADAMS, , V.L. EHLERS, J.R. MOYER, JR. University of Wisconsin-­‐Milwaukee Stroke is one of the leading causes of death and disability in the United States, costing the country $312.6 billion each year (Go et al., 2013). During an ischemic insult, brain cells are deprived of oxygen and glucose, which leads to excessive depolarization and calcium (Ca2+) influx, and in turn, activation of cell death cascades and eventual cell death. Endogenous Ca2+-­‐binding proteins (CaBPs) limit ischemic damage by buffering excess Ca2+. Apoaequorin (AQ) is a Ca2+-­‐binding photoprotein found in the jellyfish Aequorea victoria. In previous experiments, our lab has shown that a single intrahippocampal infusion of AQ 24 and 48 hours prior in vitro ischemic insult significantly reduces cell death (Detert et al., 2013). It has also been found that there are concurrent changes in cytokine mRNA after AQ infusion, including interleukin-­‐10 (IL-­‐10) and tumor necrosis factor-­‐alpha (TNF-­‐α; Detert et al., 2013). These data indicate that AQ’s neuroprotective mechanism may involve modulation of certain anti-­‐ and pro-­‐inflammatory molecules, possibly involving a preconditioning-­‐like effect. The current study was designed to further investigate whether cytokine protein expression also changes in a time-­‐dependent manner after an intrahippocampal infusion of AQ. By focusing on possible changes in protein levels, we hope to gain a better understanding of the extent to which AQ modulates various cytokines and ultimately understand the mechanism by which AQ protects neurons from oxygen-­‐glucose deprivation. Grant/Other Support: CalciGenix 83 Kyle Minnick Carlita Favero Effects of moderate prenatal alcohol exposure on cells of the ventral telencephalon K. MINNICK, T. OLAFUNMILOYE, S. WHITE, B. HENSTENBURG, C. FAVERO Ursinus College Individuals affected by Fetal Alcohol Spectrum Disorder (F ASD)experience limited development of the central nervous system, specifically in higher brain function. Thalamocortical axons (TCAs) connect the thalamus and the cortex of the brain and are involved in our ability to sense and perceive and respond to everything that is going on around us. For these axons to form correctly, they are guided through the ventral telencephalon by corridor cells. Previous studies show that cells similar to corridor cells are affected by prenatal ethanol exposure (Zhou et al. 2005), so we are interested in studying these potential effects and how they may later influence development of TCAs. For these studies mice drank either 20% ethanol or tap water for 2 hours a day from embryonic day 1 to 14. We euthanized pregnant females and removed embryos by C-­‐section on day 14. We studied these brains using two methods of staining, Nissl 58 stain and Islet 1 and Pax6 immunostaining. Nissl staining allowed us to determine the approximate region of the brain section, while Islet 1 transcription factor showed the cells making up the corridor. Pax6, a transcription factor, Constrains development of the corridor. We saw fewer islet1 cells and more Pax6 cells in the ethanol exposed brains. Though this is preliminary data, we hypothesize that there is likely a problem with cell specification or migration caused by alcohol exposure. Email: [email protected] 84 Alexander Ordoobadi Josef G. Trapani Hair cell encoding of intensity in the zebrafish lateral line A.J. ORDOOBADI, R. AZIZ-­‐BOSE, J.G. TRAPANI Amherst College Hair cells, the sensory receptor of the auditory and vestibular systems, quickly and precisely encode various properties of mechanical stimuli, including intensity and frequency, into trains of action potentials (spikes) in afferent neurons. While much is known about the mechanisms of hair cell encoding, there are still many unanswered questions related to how hair cells encode stimulus intensity. Here, we used an in vivo larval zebrafish preparation to study how hair cells encode the intensity of mechanical stimuli. Briefly, a waterjet was used to mechanically stimulate the lateral line hair cells of a larval zebrafish, and the resulting trains of action potentials were recorded from the cell body of an innervated afferent neuron. By varying waterjet pressure, we found that the latency to the first elicited spike (FSL) decreased as stimulus intensity was increased. In addition, with repeated stimulation, the variability in the time to the first spike decreased with increasing stimulus intensity. FSL is an important encoding scheme because it provides a rapid means of conveying information about a stimulus, but it can only be accurately interpreted as a feature of encoding if there is high fidelity in the first spike times. To further elucidate how hair cells encode intensity, we used a transgenic line of zebrafish that express Channelrhodopsin-­‐2 (ChR2), a light-­‐gated ion channel, in hair cells of the ear and lateral line. Using this transgenic line, we depolarized hair cells with light, bypassing the hair cell’s mechanotransduction (MET) channel. Interestingly, hair cells stimulated with light also showed a decrease in FSL with increasing light intensity, indicating that the MET channel is not essential for the encoding of intensity by FSL. However, preliminary results suggest that the fidelity of FSL during repeated stimulation with light was not as robust as with mechanical stimulation. Together, these results suggest that hair cells in the zebrafish lateral line encode stimulus intensity in the timing of the first action potential elicited in the afferent neuron. Email: [email protected] 85 Sally Curtiss Sarah Blythe The effects of diet-­‐induced obesity and gender on the morphology of hippocampal neurons S. CURTISS, C. GLASS-­‐WALLEY, N.GUNAWANSA, M. KNABE, S. MARWITZ, K. SARFERT, L.WOODIE, S. BLYTHE Washington & Lee University Throughout the United States more than one-­‐third of adults are obese. Obesity is associated with a number of serious health problems including heart disease, insulin insensitivity, hyperlipidemia, and certain types of cancer. Moreover, recent studies have shown that the intake of diets high in fat, sucrose, and fructose, can lead to cognitive impairments. Researchers have also found that estrogen has major neuroprotective benefits. Estrogen has been seen as a beneficial treatment to reduce the risk of Alzheimer’s disease, reducing the risk of about 29-­‐44%. But, estrogen has not shown any difference in ameliorating the decline of cognitive function in women already diagnosed with Alzheimer’s disease. To 59 study the effects of estrogen protection in an obese rat model, male (n=14) and female (n=20) Sprague-­‐
Dawley rats were placed on either a western-­‐style or control diet for 10 weeks. All female rats were ovariectomized (OVX), and half were given subcutaneous 17 β-­‐Estradiol pellets to provide a low, constant level of estrogen. The Western-­‐style diet was high in fat and processed sugars and more calorically dense than a typical control chow. Rats were then deeply anesthetized and transcardially perfused with saline. Some brains (n=22) were removed and placed into Golgi-­‐Cox solution for two weeks. The tissue was then section and stained to reveal dark black cells. Candidate neurons were identified in the CA1 region and the dentate gyrus, and then imaged using confocal microscopy. The cells were then reconstructed in Neuromantic, an open source neuron reconstruction tool. The neurons will be evaluated for branch length, number of dendritic branches, spine density, and overall dendritic morphology. The remaining brains (n=12) were maintained in paraformaldehyde and were later sectioned and stained for GFAP and Doublecortin in order to assess glial proliferation and neuronal precursor cells. Statistical analysis was performed between dietary and hormonal groups. We found that neurons in the CA1 region of the hippocampus had significantly greater branch points and terminal tips in male rats in comparison to both OVX and OVX+E females. Because of the difference observed in the CA1 pyramidal cells but not in the Dentate Gyrus cells we believe there is an issue of access of memories rather than an issue of actually making memories. Email: [email protected] 86 Melina Knabe Sarah Blythe Cafeteria-­‐style diet impairs memory and increases risk of metabolic syndrome M. KNABE, K. SARFERT, S. CURTISS, C. GLASS-­‐WALLEY, S. MARWITZ Washington & Lee University Increasing evidence suggests that excess energy intake and obesity are associated with cognitive dysfunction (F. Berrino, 2002; Kanoski and Davidson, 2011). Indeed, prolonged exposure to diets high in saturated fat and simple sugars contributes to the development of neurological disorders such as Alzheimer’s disease and ADHD (Miller et al., 2014; Waring and Lapane, 2008). In order to study the effect of diet-­‐induced obesity on physiology and behavior, multiple diet models exist. Although the high fat/high carbohydrate Western-­‐style diet is often used to study diet-­‐induced cognitive impairment, we believe the cafeteria-­‐style diet (CSD) more accurately represents the varied and energy-­‐dense diet that contributes to hyperphagia and obesity in humans. The CSD is low in fiber and nutritive value, and it also offers a range of flavors and textures, which increases its hedonic appeal. Therefore, the aim of this study was to investigate the physiological and behavioral effects of the consumption of a CSD on juvenile rats. In the present study, 22 male Sprague-­‐Dawley rats were divided into control (n=11) and CSD (n=11) groups at weaning. Diet exposure continued for 10 weeks, during which all animals were given ad libitum access to standard rat chow and water. The CSD animals were simultaneously offered energy-­‐dense foods in the form of two alternating diets. These diets included various cookies, chips, processed meats, and sweetened drinks. During weeks eight and nine of diet exposure, the rats were subjected to Novel Object Recognition, Novel Place, Novel Context, and Morris Water Maze tasks to assess various forms of memory, as well as an open field task to assess locomotor behavior. Following sacrifice, fat pads, livers, aortas, and brains were excised for post-­‐mortem studies. We did not see the expected bifurcation in weight gain between control and CSD groups; however, CSD rats had significantly higher abdominal and gonadal fat pad weight than the control rats (Student’s t-­‐test, p<0.05). There were no signs of the development of non-­‐alcoholic fatty liver disease, although the percentage of total aorta area composed of lipids was slightly elevated in CSD rats. Our study also demonstrated that rats fed a cafeteria-­‐style diet had compromised spatial and episodic memory, as indicated by Morris Water Maze and Novel Object Recognition tasks (Student’s t-­‐test, p<0.05), but they did not exhibit hyperactive behavior. These data suggest that prolonged exposure to an energy-­‐dense CSD leads to memory impairment and the replacement of lean muscle with abdominal fat. These two effects may contribute to the development of metabolic syndrome and neurological disorders. This work was supported by the following sources: 60 Virginia Academy of Sciences Undergraduate Research Award, Summer Research Scholar Program, Lenfest Faculty grant, and W&L HHMI Training Program. Email: [email protected] 87 Varun Wadia Richmond Thompson The role of ultraviolet reflectance in female mate choice in goldfish V. WADIA, E. KIM, C. ANDERSEN, J. GOMEZ, D. GONZALES, R. THOMPSON Bowdoin College The recent discovery that steroid hormones have rapid effects on responses to visual stimuli in goldfish, Carassius auratus, has led to a number of studies attempting to find the mechanism of these rapid effects. It is thought that the hormones may be modifying visual processing in the fish to enhance behavioural responses to these cues. The focus of this study was therefore to determine which visual cues are important to the fish in their sexual interactions. Goldfish do not have sexually dimorphic reflectance patterns in the visible range, but they can detect ultraviolet (UV) wavelengths with a cone type that is maximally responsive to 356 nm, suggesting sexually dimorphic UV reflectance could be used during sexual interactions in this species. To test this, we performed behavioural approach experiments in varying intensities of full-­‐spectrum light in which females were given a choice between two males, one from which the reflected UV light was blocked by a Plexiglas barrier and one from which UV wavelengths could pass through a special UVT Plexiglas barrier. Olfactory cues were blocked by both barriers. Females spent significantly more time near males behind barriers that allowed UV transmission than near males behind barriers that did not, but only in tests in which females were treated with a hormone that induces receptivity and only if a male pheromone (androstenone) was also added to the water. These results suggest that UV reflectance patterns may be an important visual cue for female mate choice in this species. Funding sources: National Center for Research Resources (5P20RR016463-­‐12) and the National Institute of General Medical Sciences (8 P20 GM103423-­‐12) from the National Institutes of Health, NASA. Email: [email protected] 88 Emily Sewell David M. Hollis Effect of CSPG inhibitory peptides on dorsal root ganglion neurite growth E.C. SEWELL, Y. OTAKE Furman University Chondroitin sulfate proteoglycans (CSPGs) are a family of extracellular matrix molecules that play a key role in neuronal development and glial scar formation. They are highly upregulated after CNS injuries in the glial scar and they mediate neuronal growth suppression. In previous studies, the drug Chondroitinase ABC has been found to inhibit CSPG function; however, it cannot cross the blood-­‐brain barrier and has a short period of enzymatic activity. In this study, we identified inhibitory peptides (CDPs) to overcome inhibition by CSPGs to facilitate CNS axon regeneration more efficiently. CDP peptides enhanced DRG neurite outgrowth on aggrecan spots in vitro. Specifically, CDP peptides 2,3,4,6,7,8, and 9 were able to attenuate CSPG activity and promote axonal growth. Identification of CDP peptides as novel CSPG inhibitors should facilitate the development of an applicable drug to promote axonal regeneration. Research Support: Furman Advantage Shriners Pediatric Neural Rehabilitation Center Email: [email protected] 61 89 Madeleine Searles Jeffrey S. Smith Does acutely placed embryonic neural stem cell therapy induce restoration of function following cortical contusion impact in adult rats reared in an enriched environment? M.A. SEARLES, I.J. HIATT, K.A. MEERSCHAERT, J.S. SMITH The Brain Research Laboratory, Saginaw Valley State University Traumatic brain injury (TBI) is among one of the leading causes of disability and mortality. Each year approximately 1.7 million individuals sustain a TBI. Currently, no effective clinical treatments exist to combat functional and cognitive deficits incurred from TBI. Recent research has shown that enriched environments (EE) aid in functional recovery following TBI. Embryonic neural stem cells (eNSC) express neuronal characteristics and have been shown to survive, migrate after transplantation, and improve recovery after functional loss. Furthermore, recent data from our lab has shown that combining EE and eNSC therapy improves recovery from TBI and appears to increase the survival rate of eNSCs. The purpose of the current study was to examine the influence that acute transplantation of eNSCs, post-­‐
injury, may have on eNSC survival, neuroplasticity, and overall functional recovery following a MFC contusion in rats reared in an EE. Thirty, twenty-­‐five day-­‐old male Long-­‐Evans rat pups were reared in EE housing. After ninety days in the EE, twenty animals received a contusion injury to the MFC. Seven days post-­‐injury, half of the injured animals were transplanted with ~100,000 eNSCs in two locations near the lesion. Behavioral analysis was conducted using the open field task (OFT), Barnes maze (BM), Morris water maze (MWM), rotor-­‐rod (RR), elevated-­‐plus maze (EPM), and the forced-­‐swim task (FST). Following behavioral testing the animals were euthanized, perfused, their brains extracted, and the tissue was imbedded in paraffin, sectioned, and underwent hematoxylin and eosin staining. Stereological analysis was performed to quantify number of surviving cells and total cortical volume. The data suggests that injured animals who received eNSCs performed better during the MWM task, when compared to injured animals who did not receive eNSCs. For the RR task, the data shows no differences between the treated and non-­‐treated injured groups, though both were significantly impaired when compared to intact controls. The current data suggest that timing of placement into the enriched environment may affect the performance of eNSC therapy and should be explored further. Email: [email protected] 90 Jacob Dunkerson Jeffrey S. Smith Does chronically placed embryonic neural stem cell therapy induce restoration of function following cortical contusion impact in adult rats reared in an enriched environment? J.A . DUNKERSON, M.A. SEARLES, D.A. FOSTER, K.M. LIZYNESS, N.P. TAUSCH, J.L. WALKER, J.S. SMITH The Brain Research Laboratory, Saginaw Valley State University, University Center, Michigan Despite advances towards potential clinically viable therapies there has been only limited success in improving functional recovery following traumatic brain injury (TBI). Exposure to an enriched environment (EE) improves memory, learning, and motor skill development in both intact and injured animals. Embryonic neural stem cells (eNSCs) have been shown to survive transplantation and influence acute functional recovery in various models of acute TBI. Though the combined therapies have improved acute recovery, they have not been tested as a therapy for the chronic symptoms of TBI. Based on the current evidence it is hypothesized that transplantation of eNSCs 42 days following a medial frontal cortex (mFC) controlled cortical impact (CCI) injury will significantly improve functional recovery. The subjects of the study were thirty adult male Long-­‐Evans rats, which were raised in an EE from post-­‐natal day 25 to adulthood. At approximately 90 days of age, 20 adult male rats were randomly selected to receive a midline mFC cortical contusion impact (CCI) injury. The injured rats were individually housed in a 62 standard environment (SE) for seven days before being randomly reassigned back into an EE. At 42 days post-­‐injury 10 injured rats were randomly selected to receive bilateral transplantation of 100,000 eNSCs and immediately returned to their EE. Behavioral analysis, which included the Open-­‐field test, Morris water maze (MWM), Rotarod (RR), Elevated plus maze, and Forced swim task, was conducted over 30 days, beginning seven days post-­‐transplantation. The current findings of the study suggest that there were no significant differences between the TBI-­‐eNSC and TBI-­‐Untreated groups on the MWM task or the Rotorod task, however both groups performed significantly worse than the Intact-­‐Shams. These findings indicate that there is a therapeutic ceiling effect associated with eNSC therapy on functional recovery. Email: [email protected] 91 Monica Gentchev Jennifer Taylor The effect of beta amyloid positivity on cerebral metabolism in cognitively normal seniors M.K. GENTCHEV, A. BOZOKI, D. ZHU Michigan State University Alzheimer’s disease has a complicated pathology that is associated with a dramatic cognitive decline. Pathological factors and changes that are linked with the disease include, but are not limited to, beta-­‐
amyloid, apolipoprotein E (APOE), and glucose metabolism. Amyloid plaques are distributed throughout the cortex in Alzheimer’s disease (Braak & Braak, 1991). These plaques deposit in the neocortex, entorhinal cortex, insular cortex, and hippocampus, respective to disease progression (Thal, Rob, Orantes, & Braak, 2002). Apolipoprotein E is a class of apolipoprotein that is responsible for transporting lipoproteins, fat-­‐soluble vitamins, and cholesterol throughout circulation (Farlow et al, 2004). However, this transporter has an allele, APOE4 that has been associated with Alzheimer’s disease (Strittmatter et al, 1993; Deary et al, 2002). Additionally, the pathology has been associated with a decreased metabolism in the temporo-­‐parietal cortex. Recent studies have been focused on presymptomatic pathology. The goal of these studies, in addition to our own, is to find signs of decline prior to disease onset. Having a prodromal diagnostic criteria will enable clinicians to effectively treat patients, when the resources are available. Some studies, as of late, have looked into the early metabolic changes in health senior subjects. However, results have contradicted one another (Cohen et al, 2009; Vaishnavi et al, 2010; Oh et al, 2012; Drzezga et al, 2011). The contradictory outcomes may be a power problem due to low subject pool. Our study looked into the changes that occur in metabolism with respect to amyloid deposition and APOE4 status. Subject data was acquired from the Alzheimer’s Disease Neuroimaging Initiative (ADNI). Magnetic resonance images (MRI), fludeoxyglucose positron emission tomography (FDG-­‐PET) scans, florbetapir PET (AV45) standard uptake value ratios (SUV/R), and APOE4 status were acquired for each subject. MRI and FDG-­‐
PET scans were coregistered and processed to obtain a SUV for glucose metabolism in the entorhinal cortex, amygdala, hippocampus, isthmus cingulate, and medial temporal region for each patient. These SUVs were compared against the AV45 values to determine a correlation between glucose metabolism and amyloid deposition. Individuals were then grouped by APOE4 status; the values were then further analyzed. Our results show that a hypermetabolic effect is present in individuals who have higher cerebral amyloid concentration, but only in consequence of APOE4 status. These conclusions are representative of the significant effect that apolipoprotein E4 has on the medial temporal region’s metabolism. Email: [email protected] 92 Dylan Miller Laura Symonds Shocks and stings: microstimulation of scorpion defensive behavior D. W. MILLER, G. GAGE, A. ROWE Michigan State University 63 A scorpions’ experience of the world is primarily dependent upon its sense of touch, much of which is localized to sensory organs on their legs. Previous studies on scorpions have indicated that receiving direct tactile stimulation to their legs induces a defensive behavior-­‐either stinging, or snapping with their pedipalps, in the perceived direction of the stimulation, or movement directly away from the perceived stimulus. In this study, different species of scorpions had their defensive responses evoked and examined. The scorpion surgery includes interfacing implanted wire electrodes with the leg nerves, which are then paired with an external function generator. The scorpions received 2.5 V of 55 Hz AC for 150 ms, at differing legs, and their resultant behavior relative to the point of stimulation was recorded. The prediction is that those scorpions with thicker pedipalps will reliably sting or claw to the side the electrical stimulation is provided, and those with thinner pedipalps. Little is understood about scorpion defensive behavior and neurophysiology, and this is being used as a means to explore both topics. Support Provided by: Backyard Brains. LLC MSU Federal Credit Union Video: http://bcove.me/qj3elv9k Article: http://www.popularmechanics.com/science/health/nueroscience/the-­‐robo-­‐scorpion-­‐is-­‐coming-­‐
and-­‐no-­‐one-­‐is-­‐safe-­‐17035113?click=pm_latest Email: [email protected] 93 Cort Thompson Laura Symonds Undergraduate optogenetics researcher C. H. THOMPSON, O. SHAFER, G. GAGE Michigan State University Optogenetics enables the manipulation of neural activity in free moving organisms with millisecond precision by making modified ion channels sensitive to a particular wavelength of light. Unfortunately today, most optogenetics methods are expensive and out of reach beyond well funded institutions. This is regretfully common within the field of neuroscience and most students aren't exposed to any neuroscience concepts until graduate education. Making neuroscience methods such as optogenetics available in grade-­‐school classrooms is important for introducing students to the excitement of neuroscience. Neuroscience is a rapidly growing field and bringing the most cutting edge methodologies, such as optogenetics, to the classroom will play a role in expanding the field of neuroscience in the future. Using a red-­‐shifted opsin (ReaChR) that has been developed to allow light to penetrate through the exoskeleton of insects and activate target cells, I will develop protocol and experiments using affordable tools and materials to observe which neural pathways are involved in the Proboscis Extension Response(PER). Creating new tools for understanding the systems behind animal behaviors is important not only because it can inspire interest in neuroscience and encourage critical inquiry in youths, but to eventually gain a further understanding of the mechanisms of neural substrates similar to those of humans via such animal models. Email: [email protected] 94 Amy Pritchard Dawn M Blitz Neurohormonal modulation at neuromuscular junctions exhibiting distinct plasticity A.E. PRITCHARD, D.M. BLITZ Miami University Central pattern generator (CPG) circuits produce motor outputs underlying rhythmic behaviors such as chewing and locomotion in vertebrates and invertebrates (Briggman and Kristan 2008, Annu Rev Neurosci). Motor neurons relay CPG patterns to muscles via synapses known as neuromuscular junctions (nmjs). One motor neuron can synapse onto multiple muscles which can respond to the same motor 64 neuron activity differently (Katz et al. 1993, J Neurosci). Distinct responses can be elicited due to differences in synaptic plasticity such as augmentation, summation, facilitation, and depression (Zucker and Regehr 2002, Annu Rev Physiol; Stein et al. 2006, J Exp Biol). In this project, we use the stomatogastric nervous system of the crab Cancer borealis to determine if different muscles innervated by the same motor neuron which exhibit distinct amounts of augmentation also exhibit distinct levels of other forms of plasticity. Additionally, we test the hypothesis that a neurohormone with central actions on motor neuron activity also alters motor output by modulating plasticity at peripheral nmjs. Muscles gm5b and gm6ab are innervated by the LG motor neuron, a gastric mill (chewing) CPG neuron. These muscles experience distinct amounts of augmentation in response to gastric mill-­‐like LG bursts (Pritchard and Blitz, unpublished). We now find that both muscles also exhibit facilitation analyzed using paired-­‐pulse LG stimulation (interstimulus intervals: 50ms-­‐10s) and that similar to augmentation, facilitation was greater in gm6ab (ex: 50ms, p=0.011; gm5b: n=8; gm6ab: n=6). To determine whether a neurohormone which alters LG spiking activity (CCAP; Kirby and Nusbaum 2007, J Neurophysiol) also modulates nmj plasticity properties, we repeated paired-­‐pulse stimulations in CCAP. Although CCAP increased gm5b EJP amplitude (n=8, p=0.004), there was no difference in EJP paired-­‐pulse ratios in control vs. CCAP in gm5b (n=8; p>0.05) or gm6ab (n=6; p>0.05). Given the additional complexity of summation and augmentation during bursts of activity, we examined muscle responses to gastric mill-­‐like LG bursts in saline vs. CCAP. However, unlike saline conditions, EJP bursts in CCAP elicited movements (gm5b, n=6; gm6ab, n=4) which hindered quality recordings suggesting a CCAP effect that we were unable to measure. Thus, we next aim to address whether CCAP modulates augmentation during bursts and/or modulates contraction mechanics via postsynaptic mechanisms using tension recordings to further understand the interplay between synaptic plasticity and hormonal modulation at central and peripheral sites. Support: NSF IOS-­‐
1153417 (DMB) and Miami University USS (AEP). Email: [email protected] 95 Susan Rus Leslie Kwakye The neural correlates of multisensory temporal processing in an audiovisual steady-­‐
state electroencephalogram task. S. RUSS, E. ALIGBE, M. JOHNSON, L. KWAKYE Oberlin College Neuroscience Department Many studies have investigated the behavioral consequences and neural correlates of temporal multisensory processing in humans as well as animal models. Collectively, these studies demonstrate the necessity of the precise temporal alignment of multisensory stimuli for multisensory integration. These studies have also revealed a fundamental network for assessing the relative timing of audiovisual stimuli that includes the visual cortex, auditory cortex, and superior temporal sulcus. One caveat is that almost all of these studies have utilized discreet auditory and visual stimuli. Many naturalistic audiovisual objects produce somewhat continuous stimuli in the auditory and visual modalities that feature regularly repeating elements. The synchrony of these stimuli may not be judged by differences in onset as is the case with discrete audiovisual stimuli. Instead, individuals may rely on the overall temporal pattern produced in each modality. In the current study, we took advantage of the ability of sensory neurons to entrain to regularly repeating elements of external stimuli to produce steady state visual and auditory potentials. Visual stimuli consisted of a white square that flickered at a rate of either 10Hz or 30Hz. Auditory stimuli consisted of amplitude-­‐modulated white noise at frequencies between 4Hz and 48Hz. The audiovisual stimuli were synchronous (both visual and auditory frequencies at 10Hz or 30Hz), or asynchronous, with the auditory being faster or slower (20% or 60% increase/decrease). Auditory and visual-­‐only stimuli were presented with static stimuli in the other modality. Participants were asked to report whether the visual and auditory stimuli were synchronous while continuous electroencephalograms were recorded using a 64-­‐channel ActiChamp system. Preliminary behavioral data indicate that participants were generally more accurate at identifying the synchrony of audiovisual stimuli centered at 10Hz as compared to 30Hz. Additionally, participants were more accurate at correctly identifying asynchronous audiovisual presentations when the auditory stimulus was presented at a 65 slower rate than the visual. Future analyses will investigate changes in oscillatory amplitude and coherence for trials in which participants reported the audiovisual stimuli to be synchronous as compared to asynchronous. Additionally, we will investigate individual differences in the mechanism for multisensory pattern discrimination that may be applicable to the study of individuals with known alterations of multisensory temporal processing (e.g. in autism spectrum disorders or in musicians). Email: [email protected] 96.1 Dan Luu Sarah Webster Measuring behavioral response to stimulation of the bristles in Drosophila melanogaster D. D. LUU, S. M. WEBSTER College of the Holy Cross Mechanosensation is the process of interpreting physical stimuli like touch, sound, or vibration from one’s surroundings. The fruit fly bristle is similar to the human hair cells in the inner ear, making Drosophila melanogaster a useful model study for human hearing and balance. We can measure mechanosensation by observing a natural behavior when flies initiate a grooming reflex in response to the touch of a bristle. In preparation to study the grooming behavior in mutant flies in the future, we created mosaic flies with a subset of mechanosensory bristles marked by green fluorescent protein (GFP). We initiated a grooming response by stimulating the marked bristle with a light touch and scored the grooming response. After testing eight different types of bristles, we found that the best responders are flies tested on the post-­‐alar bristles. The post-­‐alar bristles responded 40% of the time when stimulated once every two minutes for five times. Post-­‐alar bristles are also useful because, even in the lower percentage response trials, the flies responded at least once in every trial. By screening different types of fly bristles from a control fly, we were able to establish a baseline for future testing of mutants. Our next step is to test the grooming reflex in a collection of mutant mosaic flies to study whether mutations affect the mechanosensory pathways. We would like to thank The Renee and Anthony M. Marlon, M.D. '63 Summer Research Fellowship Program and the Department of Biology for financial support. Email: [email protected] 96.2 Sarah Smith Sarah Webster Wasabi aversion in Drosophila adults and larvae S.E. SMITH, S.M. WEBSTER College of the Holy Cross A variety of substances that are identified as bitter and pungent by humans can also be detected via chemosensation by the common fruit fly, Drosophila melanogaster. Wasabi is made pungent by the presence of isothiocyanate (ITC). ITC is detected by the TRPA1 ion channel, which is known to be involved in other sensory neural responses and mediates pain, irritation, and inflammation. The TRPA1 channel can be found in both humans and Drosophila, which makes the fruit fly an excellent model organism for better understanding the mechanism of this chemical detection system. As there is little published data addressing the behavioral effects of exposure to ITC in Drosophila larvae, my research aimed to explore this further. I utilized three different assays to measure feeding preference and aversion in larvae and one assay to measure these behaviors in adult flies. The known aversive substance, quinine, was used as a control for assessing the ability of the various assays to measure avoidance behavior. As other groups have reported previously, I also observed that adult flies avoid ITC when given a choice. Interestingly, as the fly starvation time increases, the aversive behavior towards ITC decreases. Conversely, larvae do not exhibit an initial aversion or feeding preference in the presence of either ITC or quinine. Because it has been previously demonstrated that larvae avoid quinine, we believe our assays are not a good tool to measure food aversion. We plan to modify the experiments by staging the larvae more precisely, as well as 66 testing larvae throughout different stages of development. Once this is established, my research will aim to examine if TRPA1 influences larval avoidance, and if so in which cells it is required. Email: [email protected] 97 Shayna Sosnowik Joshua Brumberg Morphological characterization of supragranular neurons in the primary somatosensory cortex S. SOSNOWIK, J.C. BRUMBERG, C.H. TSE, A. TSIMOUNIS Queens College-­‐CUNY The cerebral cortex is essential for cognitive computations, such as the movement of a limb or the detection of objects on the skin surface. The processors of the cortex are individual neurons and the circuits in which they are embedded. It has been shown that specific morphologies are correlated with specific circuit functions. We performed a morphological analysis of layer 2/3 neurons in the barrel cortex of the mouse as an approach to decipher the neuronal circuit(s) in this region of the primary somatosensory cortex (S1). Our aim was to objectively determine if there are morphological characteristics that can distinguish one group of neurons from another. Individual neurons from the barrel cortex in brain slices from CD-­‐1 mice were labeled non-­‐selectively with DiI using biolistics and reconstructed three-­‐dimensionally from confocal image stacks. Morphological parameters of cell bodies and dendrites of supragranular neurons in the barrel cortex were measured. Cluster analysis following principal component analysis of the morphological parameters revealed distinct groups of neurons. In order to assign “functionality” to the groups in vivo injections of fluorescent beads in the ipsilateral M1 were used to label S1 neurons that projected to M1 (a known target of supragranular barrel cortex neurons). Acute sections from these animals were processed with biolistics as described above. These experiments reveal the distribution of M1-­‐projecting supragranular layers in the overall classification dendrogram and show the correlation between anatomical classes of neurons and specific role(s) within the cortical circuit. The present results further support the hypothesis that neurons involved in specific anatomical pathways have unique morphological properties Support: PSC-­‐CUNY Award #65323-­‐00 43, PSC-­‐CUNY Award #66224-­‐00 44 Email: [email protected] 98 Stuart Behling James Demas Retinal circuitry mediating navigation in freshwater turtle hatchlings S. BEHLING, C. HELLER, S. FREEDBERG, J. DEMAS St. Olaf College In freshwater turtles, such as the common snapping turtle (Chelydra serpentina), the navigation of hatchlings towards water is crucial to survival. If hatchlings take too long to get from their nest to a body of water they are likely to die from predation or desiccation. Previous studies have shown that snapping turtles primarily use light cues in order to orient towards the water upon hatching. The retinal substrate for this phototactic behavior is not well understood. However, in newborn mice, which lack functional rods and cones, phototactic behavior is mediated by intrinsically photosensitive retinal ganglion cells (ipRGCs), a recently discovered class of photoreceptors. These findings raise two questions: are ipRGCs present in the retinas of turtles? If so, do ipRGCs mediate hatchling navigation? Using a multi-­‐electrode array we performed extracellular recordings which measured the response of turtle RGCs to light stimuli. In order to identify putative ipRGCs, we used a neurotransmitter antagonist cocktail to blocks rod and cone derived synaptic input onto RGCs. A small subset of the RGCs maintained the ability to respond to light in the presence of the antagonist cocktail. The response properties of the putative turtle ipRGCs 67 resembled those of mammalian ipRGCs: after a significant delay (>0.5s), cells elevated their firing rate in response to light onset, and this increase in firing rate persisted for tens of seconds. Furthermore, the light responses in these putative turtle ipRGCs were sensitive to opsinamide. Opsinamide is an antagonist of melanopsin, the photopigment responsible for initiating intrinsic light responses in mammalian ipRGCs. In sum, these results suggest that the common snapping turtle has ipRGCs similar to those found in mice. In addition, we were able to show strong phototactic behavior of hatchling snapping turtles in a laboratory setting using a Y-­‐maze. Preliminary data suggest that the phototactic behavior is more sensitive to blue light, consistent with a melanopsin-­‐based photoreceptor. In the future, we will be able to combine this behavioral assay with pharmacology in order to test whether ipRGCs can drive phototaxis independently of rod and cone mediated signals. Email: [email protected] 99.1 Joseph Schultz Karen Brakke, PhD Changes in VGLUT1 synaptic connectivity and the presynaptic inhibitory inputs that modulate their activity on motoneurons following peripheral nerve crush J. SCHULTZ, T. ROTTERMAN, A. DWARAKANATH, F. ALVAREZ Emory University Group IA afferents are pseudo-­‐unipolar sensory neurons that constitute the muscle spindles in the periphery, where they detect changes in muscle length. They relay this information to the spinal cord, where they make excitatory synapses on motoneurons, and are responsible for inducing the stretch reflex. Previous work in cats has shown that 30% of IA afferents fail to reinnervate muscle spindles after nerve crush. This is reflected in 30% reduction in the amplitude of synaptic potentials elicited by muscle stretch on motoneurons (Prather et al., 2011). These same cats, however, demonstrate supranormal generation of force during stretch reflexes elicited after regeneration of these crushed nerves (Prather et al, 2011), suggesting that somehow transmission between IA afferents and motoneurons is facilitated during the stretch reflex. This phenomenon might be explained by a loss in presynaptic inhibition, via GABAergic control, of the IA afferent synaptic contacts on motoneurons. We therefore analyzed the density on injured and regenerating motoneurons of IA afferent synapses in adult female Wistar rats detected immunocytochemically through their content of vesicular glutamate transporter 1 (VGLUT1) and their presynaptic inhibition from GABAergic synaptic terminals expressing the 65 kDa isoform of glutamic acid-­‐
decarboxylase (GAD65). 99.2 Deanna Ross Karen Brakke, PhD Detection of ephrin-­‐A5 in the superior colliculus of mice D. ROSS, V. CLARK, S. PALLAS Georgia State University This research is focused on understanding the molecular mechanisms that aid in plasticity after damage to the visual system. In the visual system, Retinal Ganglion Cells (RGCs) send information to the superior colliculus (SC). Our lab uses a model for brain trauma in which the caudal SC is damaged at birth in hamsters. After the damage, the RGCs compress their retinotopic map onto the residual SC. Previous studies in mice and hamsters found that a class of axon guidance molecules called ephrins and their Eph receptors are expressed in a graded fashion in the SC and play a role during development. After neonatal partial tectal (PT) ablation in hamsters, both the retinocollicular map and ephrin-­‐A5 expression gradient were shown to compress, leading to the question whether ephrinA5 is instructing the compression. In this lab, electrophysiology is used to assay a retinoptic map on an ephrinA knockout mouse model. Results have been ambiguous. To test whether different levels of ephrinA5 protein can be detected, a western blot 68 was performed to quantify the differences in the knockouts. We find that there is not a statistical difference between a normal mouse and a mouse with an altered genotype (p > .05). 100 Kathryn Annand Dr. Victoria Turgeon Growth and imaging of a spinal cord in a three-­‐dimensional cell culture in the presence of a PAR-­‐1 activator K. ANNAND, S. BEASLEY, N. CAMP, M.L. MITCHELL, V. TURGEON Furman University 2-­‐Dimensional cell culture systems have been used for decades in studies involving cell interactions, however there are problems associated with these arrangements. The use of 2D systems is especially problematic when dealing with Schwann cells and motor neurons, as the plastic base inhibits the Schwann cells from fully myelinating the motor neuron axons. While there are 3D options available for co-­‐culturing cells, many still do not accurately represent conditions in vivo. The present study explored the viability and behavior of both motor neurons and Schwann cells in a 3D cell culture system, specifically rat tail type I collagen scaffold. There was an additional focus on how protease-­‐activated receptor-­‐1 (PAR-­‐1) activation with the amino acid sequence SFLLRNP, an activator of the PAR-­‐1 receptor, affects the cells’ growth and behavior. To begin, the rate of motor neuron axon growth through the collagen scaffold was determined in both the presence and absence of the PAR-­‐1 activator. PAR-­‐1 activation was shown to decrease the rate of motor neuron axon growth at all time periods, and it was determined that the axons grew at an approximate rate of 1.17 microns per hour through this scaffold. The effects of PAR-­‐1 activation on Schwann cell growth were simultaneously investigated. PAR-­‐1 activation caused a morphological change in the Schwann cells. Cells that were treated with the amino acid sequence SFLLRNP had observable projections coming from their normally circular cell bodies. The establishment of the motor neuron axon growth rate in the presence of the amino acid sequence SFLLRNP allows for more efficient co-­‐culturing of motor neurons and Schwann cells, as the amount of time needed for the neurons to fully extend their axons through the collagen can be calculated. The presence of SFLLRNP should facilitate Schwann cell myelination of the motor neurons’ axons in a specific 3-­‐D apparatus, which will allow future hypotheses concerning these interactions, specifically the role of the PAR-­‐1 activation on myelination, to be studied. Future studies will also be concerned with identifying specific proteins responsible for the formation of the processes mentioned. 101 Kimberly Huynh Vonnie Shields From molecules to motion: Assessing the responses of house crickets to plant volatiles using behavioral and electrophysiological paradigms K.A. HUYNH, M.C. NESLUND, T.C. BAKER, V.D.C. SHIELDS Towson University Olfaction plays an important role in the orientation and food source location for many animals including insects. Insects are capable of detecting and discriminating large numbers of odorants that differ in size, shape, and complexity. To gain insights into the mechanisms underlying odor-­‐mediated orientation, it is necessary to study how odorants are detected, discriminated, and processed in the brain. The house cricket, Acheta domesticus, bears a pair of long antennae for the detection of diverse odorants by means of olfactory receptor cells (ORCs) residing in different types of cuticular sensory organs (sensilla). These sensilla act as the first level of environmental perception and are the crucial interface between the insect’s outside world and its central nervous system. Odor molecules first reach the surface of these sensilla, perforated by many small pores, and find their way to the underlying sensory neurons where they bind to specific receptor sites. Here, the ORCs transduce the chemical stimuli into electrophysiological signals. 69 This information is sent to the brain of the insect and provides neural input for higher order processing. Subsequently, this neural processing gives rise to behavioral orientation responses. In this study, we used a Y-­‐tube olfactometer to screen a large number of host-­‐plant-­‐associated odorants, selected from a wide array of chemical classes, to determine which ones elicited positive (attractive) and negative (repellent) behavioral responses for both female and male crickets. In addition, we used an electroantennographic detection technique (EAG) to test the functional relevance of these odorants. We found some volatiles elicited strong EAG responses, while others evoked medium to weak responses. The results of these combined research approaches contribute to our knowledge of important plant odorants necessary for insect-­‐plant interactions. Email: khuynh1@students.towson.edu 102.1 Bridget Mitchell Corey Cleland Determinants of the escape response of crickets to localized heat stimuli B.C. MITCHELL, S.C. HEITSCH, G.W. REBHUN, E.G.THOMSON, R.L. GAITA, C.L. CLELAND James Madison University Animals respond to aversive stimuli with escape or withdrawal responses. In crickets, wind or looming stimuli, which might normally be produced by an approaching predator, are commonly used to evoke an escape response in which the cricket turns and then runs or jumps away. Although in mammals aversive heat stimuli have been used routinely to evoke nociceptive withdrawal responses, there have been no studies of the cricket’s response to localized heat stimuli. The goal of this study was to describe the escape response of the cricket (Acheta domesticus) to heat stimuli delivered to each of its six tarsi and determine the factors that control direction and magnitude of the response. Heat was delivered to the tarsus of each leg in 25 crickets with an infrared laser diode (980nm) and the response was recorded by high-­‐speed video (Redlake/IDT, 650 fps) placed overhead. The top of the head, thoracic-­‐abdominal junction and the tip of the abdomen were tracked over time (Proanalyst, Xcitex) to provide the two dimensional locations and orientation of the abdomen and the head/thorax. Further, the initial locations of the tarsi just prior to movement were recorded. In response to heat stimuli, crickets first retracted the stimulated tarsal, then turned by pivoting about a point toward the rear of the animal, and finally either walked (86%), jumped (9%) or remained largely stationary (5%). As with wind or looming stimuli, the turn was always away from the location of stimulus. In contrast, however, jumping was less frequent than with the other types of stimuli. The rotation of the head/thorax matched the rotation of abdomen, unlike crickets’ response to looming stimuli in which the head leads the abdomen. These results demonstrate that crickets escape from heat as well as from looming, touch and wind stimuli, and offer the opportunity to identify common movement strategies by comparing the escape responses to the four different stimuli. 102.2 Kimberly Seamon Corey Cleland Rat hind limb nociceptive withdrawal response to heat stimuli depends on initial paw posture but not stimulus location K.M. SEAMON, M. HARTMANN, C.A. CHRZAN, M.N. KABORE, K.A. MOORE, C.L. CLELAND James Madison University Rats rapidly withdraw their hind limb in response to a noxious heat stimulus applied to the plantar surface of their paw, which is an example of the Nociceptive Withdrawal Response (NWR). Previous studies in spinalized or lightly anesthetized non-­‐human mammals have shown that the direction of response depends on stimulus location. The goal of this study was to determine if stimulus location, or other factors such as initial posture or response latency, determines the direction of withdrawal in the intact, unanesthetized rat. Rats were placed on a glass plate through which an infrared laser beam (980 70 nm) was directed to heat a small (1mm) localized portion of the plantar surface of the foot. The resulting withdrawal response was recorded with three conventional camcorders (60 fps @ 1080p), one on the left, one on the right, and the third underneath the rat. From the video beneath the rat, the initial location and angle of the stimulated paw was recorded. In response to the stimulus, the rat then withdrew and rapidly (~40ms) replaced its paw on the glass, at which point the final location and angle of the paw were recorded. Rats withdrew and then replaced their paw on the glass in all possible directions. To determine if the location of the stimulus influenced response direction, the rat’s paw was stimulated in five locations (three aligned rostral-­‐caudal and three aligned left-­‐right). Unexpectedly, we found no dependence on stimulus location. However, we did notice the initial position of paw varied in both location and angle. Consequently we explored if initial position (left-­‐right and rostra-­‐caudal) and paw angle influenced final location and angle. Correlation between initial and final locations and angles did reveal a highly significant linear relationship (p<0.001), regardless of response latency. These results demonstrate, in contrast to studies in spinalized or anesthetized non-­‐human animals, that initial posture plays a greater role in the programming of the NWR than stimulus location. 102.3 Ariel Childs Corey Cleland Determinants of the escape response of crickets to looming stimuli A.M. CHILDS, K.L. REIMAN, C.R. EBEL, C.L. CLELAND James Madison University Animals respond to aversive stimuli with escape or withdrawal responses. In crickets, wind, which might normally be produced by an approaching predator, has been shown to evoke an escape response in which the cricket turns and then runs or jumps away. Looming stimuli, however, better approximate the combined stimulus modalities (wind, vision and sometimes touch) associated with attack by a predator. Nevertheless, there are a limited number of studies on the response of crickets to looming objects. The goal of this study was to describe the escape response of the cricket (Acheta domesticus) to looming stimuli delivered from each of 8 angles around the cricket. Looming stimuli were created by attaching a 3” black polyurethane ball to the end of a 12” air cylinder (45 degrees to vertical) driven by compressed air at a speed of about 90 mm/sec. The direction of “attack” was varied in 45 degree increments around the cricket. The cricket’s response was recorded by a high-­‐speed video camera (Redlake/IDT, 650 fps) placed overhead. The top of the head, thoracic-­‐abdominal junction and the tip of the abdomen were tracked over time (Proanalyst, Xcitex) to provide the two dimensional locations and orientation of the abdomen and the head/thorax. Further, the initial locations of the tip of tarsi just prior to movement were recorded. In response to looming stimuli, crickets typically first pointed their proximal antenna toward the looming object and then initiated a turn away from the stimulus. At the completion of the turn, the crickets either walked (89%) or jumped (11%) away. The direction of the turn was almost always (98%) away from the stimulus. Further, the response direction varied with the laterality of stimulus (slope = -­‐0.57; 1.0 would be directly away from the stimulus; p<0.0005). Interestingly, the head/thorax nearly always led the turn. These results demonstrate that the direction of the crickets’ escape turn from looming stimuli depends strongly on both the side and the laterality from which the stimulus is delivered. Email: child3am@dukes.jmu.edu 103 Andre DeSouza Dr. Ricardo C. Araneda Cholinergic modulation of granule cells in the olfactory bulb A. DESOUZA, R. S. SMITH, R. C. ARANEDA University of Maryland Odor information is first processed in the olfactory bulb (OB), which consists of two subdivisions, the main and the accessory OB (MOB and AOB). In both regions, output neurons of the OB, mitral and tufted 71 cells (MCs), convey odor information to other brain areas, bypassing the thalamus, highlighting the role of the OB in olfactory processing. Several inhibitory mechanisms contribute to information processing in the OB, in particular inhibition from granule cells (GCs), the most numerous interneuron of the bulb. GCs are highly regulated by both intrinsic and extrinsic signals, including cholinergic projections arising from the basal forebrain. Cholinergic modulation of GCs and MCs produces a state-­‐dependent regulation of odor discrimination. Previous work in the AOB has shown that activation of the M1 muscarinic acetylcholine receptor (mAChR) subtype produces a strong depolarization in GCs, which enhances the release of GABA onto MCs. Here, we investigated the role of muscarinic cholinergic modulation of GCs in the MOB. Surprisingly, activation of mAChRs by the non-­‐selective muscarinic agonist, oxotremorine (10 μM) produces a hyperpolarization (−6.5±0.9 mV, n=9). Furthermore, this inhibition was also present in knockout mice lacking the M1 and the M3 mAChRs. However, the selective M2 mAChR antagonist, AF-­‐DX-­‐
116 (1 μM) significantly decreased the magnitude of the hyperpolarization (−1.6±0.7 mV; n=5; p<0.05), suggesting that the hyperpolarization partially mediated by M2 mAChR. These results suggest that cholinergic modulation in the olfactory bulb is more complex than previously thought and that at the circuit level, cholinergic modulation of GC is different in the AOB and MOB. Smith RS, Araneda RC (2010) Cholinergic modulation of neuronal excitability in the accessory olfactory bulb. J Neurophysiol 104:2963-­‐
2974. Email: adesouza@terpmail.umd.edu 104 Katherine Cone Edward Bilsky Characterization of the rewarding and reinforcing effects of the mixed-­‐action delta/mu opioid agonist MMP-­‐2200 in rats K. CONE, D. GIUVELIS, J. LAVIGNE, A. LUGINBUHL, C. DUNBAR, P. ATHERTON, J. STREICHER, R. POLT, E.J. BILKSY, G.W. STEVENSON University of New England Previous work in our laboratories suggests that, compared to mu opioid analgesics, mixedaction delta/mu receptor glycopeptides might have equivalent efficacy for treating pain and reduced side effects including tolerance and physical dependence. However, the abuse liability of these compounds has not been assessed. Toward that end, this study evaluated the rewarding and reinforcing effects of a lead candidate, mixed-­‐action delta/mu agonist MMP2200. MMP2200 was first evaluated in vitro and maintained a selective efficacy for mu and delta opioid receptors versus kappa in an assay of βarrestin2 recruitment. MMP2200 was also evaluated in in vitro cAMP assays for naloxone induced overshoot and chronic morphine induced tolerance, which correlate with dependence and tolerance respectively in vivo. MMP2200 was then evaluated in vivo using a conditioned place preference assay as well as a drug self-­‐
administration procedure in rats. In place conditioning studies, rats underwent a 2-­‐week conditioning protocol and were then tested for chamber preference. Rats receiving MMP2200, at previously determined analgesic doses, could not distinguish between the drug-­‐ and saline-­‐paired chamber, whereas rats receiving the opioid agonist morphine showed a strong preference for the morphine paired chamber. In self-­‐administration studies, rats were trained to respond to the high efficacy mu opioid receptor agonist fentanyl on an FR5 schedule of reinforcement. Following complete dose response determinations for fentanyl, a range of doses of MMP2200 as well as morphine were tested. Relative to the mu agonists fentanyl and morphine, MMP2200 maintained lower rates of drug infusion. These findings suggest that MMP2200 may be less rewarding than currently available prescription opioid pain relievers. Email: kcone@une.edu 72 105.1 Ashley Lyons Brian Antonsen The role of resilin in the mantis prey capture strike A.R. LYONS, M.D. BLAKE, H.R. BREWER, R.E. THACKER, A. MUMMERT, B.L. ANTONSEN Marshall University Resilin, an elastic protein found in arthropods, has been studied in multiple high performance jumping systems. This 97% efficient protein acts as a spring, storing energy to produce quick, forceful movements, and may contribute to the remarkable speed of the Chinese Mantis prey capture strike. The work presented here is part of a multidisciplinary approach to determine if resilin contributes to muscular contraction. We performed a gross anatomical analysis to determine the distribution of resilin, from which inferences can be drawn concerning its function. We obtained calibrated photomicrographs of the strike limb segments and the joints between them from each plane. Particular attention was given to the condyles of each joint, along with the origins and insertions of the muscles. This data will be compiled into a 3-­‐dimensional computer reconstruction. Resilin was found at the attachment points of the major power stroke muscles for the strike, suggesting resilin may stretch and store energy during muscle contraction. One possible inference is that resilin acts as a transient energy storage mechanism during muscle contraction. A second possibility arising from the data is that resilin behaves as a shock absorber during the strike. A further goal of this project will be to develop a computer reconstruction using our data in order to simulate the mantis strike. Manipulation of the elastic structures will allow us to ask questions concerning our inferences of the function of resilin. Information from this project is also being used in the creation of an educational tool, which will allow students to investigate muscle and tendon kinematics. This tool is based on the Hill Muscle Model, and will contain exercises focused on lever properties, tendon stiffness and injury. Email: lyons76@marshall.edu 105.2 Anna Lefevre Brian Antonsen Neurotoxic effects of silver nanoparticles on Procambarus clarkii A.C. LEFEVRE, B.L. ANTONSEN Marshall University Silver nanoparticles (AgNP) are commonly used in medical and consumer products because of their antimicrobial properties. It has become virtually impossible to avoid coming into contact with them in day to day life since they are applied to products from children’s toys to toothpaste and shampoo. The AgNP being used in this study, and commonly in products, are coated in polyvinylpyrrolidone (PVP), which is used to prevent agglomeration and break down. They are known to cause oxidative stress in aquatic species, but the neurotoxic potential is still largely unknown. Because of the common application and lack of regulation, AgNP are inevitably in our waterways, though the exact concentration of contamination is difficult to measure. We use crayfish because they are a bioindicator and keystone species. Behavior of the organism relies on the serotonergic functioning. Based on literature, social stress influences changes in serotonergic function. Since the activity of the neurotransmitter, serotonin, on the lateral giant is well characterized in literature, we are using serotonin’s distribution on this neuron as a measure for this potential neuro-­‐toxicant. By analyzing serotonin distribution in the nerve cord, we hope to find a link between chemical stress and serotonin. We expose crayfish to concentrations varying from 1ppm to 50ppm for chronic (2 weeks) and acute (2 days) exposure periods. The nerve cord is removed and immunolabeled for serotonin, then visualized using confocal and fluorescent microscopy. Preliminary analysis indicates that treatment groups do differ from the control in location and level of fluorescence intensity. This change to the serotonergic function could cause detrimental or maladaptive effects to the behavior of the animal. Email: lefevre7@marshall.edu 73 106 Eszter Kish Michael D. Mauk Learning induces changes in the firing frequency of pons cells E.A. KISH, H.E. HALVERSON, M.D. MAUK University of Texas at Austin The pons relays the conditioned stimulus (CS) inputs to which, when paired with an unconditioned stimulus (US), the cerebellum learns to elicit a conditioned response (CR). Therefore, characterizing pontine responses to different stimuli indicates what information the cerebellum is receiving during learning. Previous studies have shown plasticity in the medial auditory thalamic nuclei (MATN) during tone mediated classical conditioning. Since the MATN has projections to the pons, plasticity induced changes in the MATN response to stimuli could influence the signals the pons relays to the cerebellum. This would indicate that although the cerebellum mediates the learning of CRs, pontine inputs dictate which CS inputs the cerebellum can learn. Pontine responses and by extension the likelihood of CRs decrease as the frequency of the stimuli get farther away from the trained frequency. This would lead to varying strengths of inputs to the cerebellum, restricting the stimuli undergoing plasticity, and directing attention to important stimuli. We tested this using tetrodes to record from the MATN and the pons in rabbits during tone and 50 ms periorbital shock (2-­‐3 mA) eyelid delay (500 ms) and trace (500-­‐500 ms) conditioning. After training to a specific frequency tone we created a session where we played a range of frequencies with only the trained frequency paired with the US. Recordings during these sessions showed a frequency curve in pons cells with higher levels of firing elicited by tones near the trained frequency. Firing levels decreased as the tones got farther away from the trained frequency. Therefore inputs to the cerebellum have different weights, as the pontine input can encode the importance of stimuli in the robustness of its firing. Performing these types of recording experiments is greatly aided by our ability to hold onto cells over different training sessions and days. This allowed us to observe changes in responses of the same cells mediated by learning over long periods of time and in different paradigms. Learning related changes to the input into the cerebellum affect how the cerebellum responds to the training stimulus. Therefore characterizing changes in pontine responses to a range of stimuli is important for understanding cerebellar mechanisms. Eyelid conditioning is performed in a lab setting where inputs to the cerebellum are controlled. In the real world however, the cerebellum receives a large range of inputs differing in importance for US prediction. Characterizing changes to pontine input to the cerebellum creates a better understanding of how the cerebellum functions in day to day situations as it encounters a spectrum of stimuli. Email: eszterkish@utexas.edu 107 Maura Curran Jacob Krans Free [Ca2+] concentration rate of decay in larval Drosophila melanogaster muscle tissue. M. CURRAN, J ARROYO, J. KRANS Western New England University Calcium [Ca2+] is vital to transduction at the neuromuscular junction (NMJ) and to muscle contraction. There remains a question of how long free calcium resides in the cytoplasm after synaptic activation of muscle during active sarcoplasmic sequestration. Fluo-­‐4 AM is a membrane permeable ester that can be used to measure intracellular calcium concentrations with high temporal and titer resolution. Here we describe the use of Fluo-­‐4 AM to estimate the [Ca2+] decay rate after multiple stimulus trains of varying parameters. We have utilized the historically important NMJ preparation of larval Drosophila melanogaster and imaged the bodywall muscles upon electrical activation. Much previous research has addressed changes in free [Ca2+] at the pre-­‐synaptic bouton rather than within the post-­‐synaptic cell (i.e. muscle). Quantification of this decay is important to understanding deviations in force generated upon stimulation by successive trains of potentials, or trains that vary stimulus rate in a non-­‐linear fashion (i.e. 74 sinusoidal activation ca. locomotion). The free [Ca2+] is estimated from the relative change in fluorescence as measured via changes in pixel intensity. Preliminary data show that fluorescence was observed to increase by approximately one hundred percent within the first train of stimulation (DF/F: 133% +/-­‐ 27%) and increase 46% (+/-­‐ 2.9%) in later stimulation trains, suggesting reduced decay rates. Indeed, after multiple successive stimulus trains, pixel intensity – and thus free [Ca2+] -­‐ decays more slowly than a when the tissue is activated with a single train. This is an important biological property as muscle contraction and force production are mediated by intracellular [Ca2+], which is released by motoneuron potential trains that oscillate between two and 35 Hz. Email: mc332191@wne.edu 108 Julia Buirkle Jennie Stevenson GHS-­‐R1A antagonism reduces alcohol but not sucrose consumption and preference in prairie voles. J.M. BUIRKLE, K.A. YOUNG, A.E. BOHIDAR, K.M. ALBERTINI, L.E. BUCKLEY, J. STEVENSON Bucknell University Rationale: Growth hormone secretagogue receptor 1A (GHS-­‐R1A) antagonism has been shown to reduce both alcohol and sucrose consumption, making the ghrelin system a potential target for the treatment of alcohol use disorders and obesity. Objective: To investigate the effects of GHS-­‐R1A antagonism in a novel animal model of high alcohol consumption, the prairie vole, and to characterize the role of ghrelin in limited access consumption of a drug (alcohol) and non-­‐drug (sucrose) reward. Methods: Female prairie voles were given four 2-­‐hr two-­‐bottle drinking sessions, occurring every other day. During drinking sessions, animals had access to 20% ethanol vs water or 10% sucrose vs water. Pre-­‐treatment with the GHS-­‐R1A antagonist JMV 2959 (i.p.; 0.0, 9.0 mg/kg Experiments 1 and 2; 0.0, 9.0, 12.0 mg/kg Experiments 3 and 4.) occurred 30-­‐min before the fourth session. To determine if the amount of exposure to sucrose sessions affected the efficacy of JMV 2959, in Experiment 5 animals were given 16 daily 2-­‐hr drinking sessions with 10% sucrose vs water. JMV 2959 treatment (0.0 or 9.0 mg/kg) occurred 30-­‐ min prior to the 16th session. Results: JMV 2959 reduced alcohol but not sucrose preference and consumption. Even after extended experience with sucrose sessions, JMV 2959 had no effect on sucrose preference or consumption. Conclusion: These findings demonstrate that GHS-­‐R1A antagonism reduces consumption of alcohol, but suggest limitations on the role of ghrelin in the consumption of naturally rewarding substances. Email: jmb081@bucknell.edu 109.1 Leigh Dairaghi Sarah H. Meerts Corticolimbic response to vaginocervical stimulation in adult female rats with or without ovaries during puberty. L.K. DAIRAGHI, N.R. MASON, S.H. MEERTS Carleton College Recent studies indicate that the female brain undergoes both postnatal and pubertal organization in the presence of ovarian hormones. Previously, we found that exposure to pubertal ovarian hormones may influence the sensory discrimination in female adult rats although sexual motivation and reward appear to develop even without pubertal ovaries. The present experiment sought to explore the role of ovarian pubertal hormones on neural activity in adult, hormone-­‐primed female rats in response to artificial vaginocervical stimulation (aVCS), which activates Fos gene expression in the hypothalamic and limbic regions of the brain. Female Long-­‐Evans rats were ovariectomized prepubertally (No O@P) or postpubertally (O@P) and then gained sexual experience at least four weeks later. On a separate day, the 75 rats were hormone-­‐primed and brain tissue was collected one hour after receipt of 15 aVCS or 15 control stimulations. The brains were sectioned coronally and analyzed immunocytochemically for differences in Fos expression within the posterodorsal nucleus of the medial amygdala (MePD), the prefrontal cortex (PFC), and the nucleus accumbens (NAc). Both the MePD and NAc are modulated by ovarian hormones and the MePD responds primarily to reproductively relevant stimuli such as aVCS. The infralimbic and prelimbic regions of the PFC are involved in activating the mating-­‐related neurons of the ventral tegmental area. MePD Fos expression was increased in female rats that received aVCS, independent of pubertal hormone condition, compared to control. However, there was no significant difference in MePD Fos expression between O@P and No O@P rats. These results are consistent with the findings of previous studies, and suggest that the neural response in female rats to sexual stimuli in adulthood is not solely dependent upon pubertal hormones. Interestingly, preliminary data show that patterns of Fos expression in the PFC and NAc are not contingent upon pubertal hormones. Our findings indicate that the adult activation of these brain regions may be independent of exposure to pubertal ovarian hormones due to the plasticity of the brain. Future studies will examine the differences in Fos expression within the anteroventral periventricular nucleus and medial preoptic area of the hypothalamus. This study was supported by an HHMI award to Carleton College. Email: dairaghl@carleton.edu 109.2 Helen Strand Sarah H. Meerts Strain differences and similarities in motivation and reward for mating and amphetamine H.K. STRAND, S.H. MEERTS, T.A. MCNAMARA, A.C. PURITZ Carleton College Female rats exhibit paced mating behavior, a pattern of approach and withdrawal from the male, when tested in an apparatus that permits the female rat to control the timing of the receipt of sexual stimulation (i.e., mounts, intromissions, and ejaculations) by exiting the compartment where the male is confined. The display of paced mating behavior varies as a function of the intensity of the sexual stimulation received from the male, such that more intense stimuli elicit longer delays in approach behavior and more withdrawals from the male. Mating is considered nonpaced when it occurs under conditions that do not permit the female to control the timing of the sexual interaction. Paced mating behavior has consistently been shown to be rewarding to female rats as measured by the conditioned place preference (CPP) task, however labs differ with regard to whether or not nonpaced mating can induce a CPP. Rat strains vary between labs and those that use Long-­‐Evans rats report that nonpaced mating is reinforcing whereas labs that use Wistar rats report that nonpaced mating is not reinforcing. A number of studies report strain differences, including strain effects in sensitivity to ovarian hormones, so we hypothesized that there may be underlying differences in response to mating stimulation that might account for the inconsistent reinforcing effects of nonpaced mating. The present series of experiments explored the response of Wistar and Long-­‐Evans rats on several tasks. Experiment 1 found that Long-­‐Evans, but not Wistar female rats exhibited a CPP for nonpaced mating. Experiment 2 found that only Long-­‐Evans rats exhibited a paced mating-­‐induced CPP. Experiment 3 analyzed behavioral measures collected during tests of paced mating behavior in sexually naïve and experienced Long-­‐Evans and Wistar rats. Sexually naïve Wistar rats showed longer return latency to the male after intromissions, fewer proceptive behaviors alone, and spent more time with the male compared to sexually naive Long-­‐Evans female rats. Wistar females displayed more rejection behaviors during both the sexually naive and experienced tests. Experiment 4 found that both strains demonstrated a CPP for amphetamine, indicating that Wistar female rats will express a CPP in our hands. Overall, these data support the idea that strain difference in response to mating may underlie the differences in the reinforcing effects of nonpaced mating. Additional experiments are required to pinpoint the neural or hormonal factors supporting these differences. 76 110 Tanner McNamara Sarah H. Meerts Conditioned object preference as a task to measure reward in adult female rats T.A. MCNAMARA, G.C. KLUMPP, S. GUTERL, S.H. MEERTS Carleton College Researchers use classical conditioning to assess the rewarding nature a range of stimuli, from drugs to mating, via the conditioned place preference (CPP) paradigm. In the CPP task a stimulus is repeatedly paired with a specific environment to determine if rats will develop a preference for that context to indicate that the stimulus is rewarding. In the present study, we asked whether Long-­‐Evans female rats would form a conditioned preference for an object associated with an injection of amphetamine relative to an object associated with an injection of saline. A tri-­‐compartment apparatus with one object placed in each outer chamber was used for conditioning and testing. A given rat received all tests and conditioning in the same apparatus. The objects in our task were a rubber truck scented with almond extract placed in the right compartment and a rubber duck scented with orange extract placed in the left compartment. Following a 15-­‐min baseline in which the rats had free access the entire apparatus, rats were randomly assigned to receive either 1 mg/kg amphetamine (experimental rats) or saline (control rats) immediately prior to 30-­‐min exposures to the compartment containing the initially less preferred object. On separate days rats received a saline injection before 30-­‐min exposures to the compartment containing the more preferred object. After three pairs of conditioning sessions, rats received a drug-­‐free test. From baseline to test, experimental rats showed a significant increase in the percentage of time spent with the object associated with amphetamine. In contrast, control rats did not show a change. To determine whether the rats demonstrated an object preference and not a place preference, a second test was conducted. The rats were tested in an apparatus other than the one in which they were conditioned and the locations of the objects were switched. Consistent with the first test, experimental rats showed a preference for the object paired with amphetamine whereas the control rats showed no change in preference. Together these results suggest that the rats associated the rewarding effects of amphetamine with the scented object, indicating that a conditioned object preference task can be used to evaluate reward. Future experiments will use this real-­‐time pairing of experience with an object to identify the specific aspects of mating that are rewarding for female rats. 111 David Liao Cheryl Harding Environmental mold exposure leads to spatial memory deficits D. LIAO, C. HARDING, R. PERSAUD, K. LIN, K. PAGE, C.L. PYTTE CUNY Hunter College An estimated 40% of American buildings contain mold. Exposure to environmental mold can cause motor impairments, chronic fatigue, and cognitive deficits. To our knowledge, no animal research has been published examining how mold causes these problems. We developed a mouse model to determine the physiological mechanisms behind these neurobehavioral issues. Adult male C57BL/6 mice were given intranasal instillations (3X/week) of 1) intact, toxic Stachybotrys spores (IN), 2) extracted Stachybotrys spores with their toxins removed and proteins denatured (EX), or 3) the saline vehicle. Our basic hypothesis is that mold inhalation activates an innate immune response, leading to brain inflammation and consequent behavioral impairment. Because innate immune activation specifically impairs hippocampal function, we examined the effects of mold exposure on performance in the Morris water maze using a two-­‐day protocol that compared the mouse’s ability to find the nonvisible platform after four visible training trials 24hr previously. As predicted, mold exposure caused significant deficits in spatial memory. After 4.5-­‐5.5 weeks treatment, EX mice performed significantly worse in finding the hidden platform compared to VEH or IN mice. EX mice showed the greatest deviation from their performances during visible training trials. They took significantly longer to reach the hidden platform and used longer paths. Performance in locating the visible platform predicted performance in locating the hidden platform 77 for VEH mice. The same was not true for the spore-­‐treated groups. Greater durations/path lengths and inconsistent performance suggest memory impairment in EX and IN mice. Performance on the water maze was inversely correlated with numbers of interleukin-­‐1β (IL-­‐1β)-­‐immunoreactive cells in hippocampal CA1 (r2 = -­‐0.56, p = 0.01). This relationship was stronger for the spore-­‐treated groups, consistent with spatial memory impairment resulting from brain inflammation due to mold treatment. Spatial memory deficits on both the last training trial (r2 = 0.52, p = 0.004) and the first testing trial (r2 = 0.59, p < 0.001) also correlated positively with weight gain during the first three weeks of treatment. Once again, this relationship is stronger for the spore-­‐treated mice. This was not surprising since energy reserves modulate immune responses. It is unclear why treatment with spore skeletons (EX) caused greater impairment than treatment with intact, toxic spores (IN). Clearly, the spore skeleton is sufficient to elicit adverse cognitive effects. These findings are consistent with brain inflammation as a cause of neurobehavioral dysfunction. -­‐1SC2 MH085472 from NIMH/NIGMS -­‐PSC CUNY Grants 69172-­‐0038 & 61214-­‐0039 -­‐Research Centers in Minority Institutions Award RR-­‐03037 from the NCRR Email: dliao711@gmail.com 112.1 Anisha Kalidindi Kyle Frantz, PhD Region-­‐specific influences of ovarian hormones on cerebral glucocorticoid receptors A. KALIDINDI, S. KELLY, C. HARRELL, G. NEIGH Emory University It is well established that stress responses can be sexually dimorphic. The glucocorticoid receptors (GR), which is present in both the prefrontal cortex and hippocampus, may be influenced by sex steroids. Therefore, understanding more about the mechanism of GR in the female rat may lead to an understanding of the sexually dimorphic nature of the stress response. Two of the co-­‐chaperones which modulate the function of the GR and are influenced by sex steroids are FKBP51 and PPID but much of what is know with their interactions are from cancer cell lines. We hypothesized that GR and its co-­‐
chaperones would be significantly influenced by ovarian hormones. In order to test this hypothesis we compared ovariectomized females to sham-­‐operated females. The target genes, GR and its co -­‐chaperones (PPID and FKBP1) were quantified through quantitative RT-­‐PCR. Removal of ovarian hormones caused a significant reduction in gene expression in the hippocampus but did not impact expression in the prefrontal cortex. These data demonstrate that sex steroids impact GR and its co-­‐chaperones in a region-­‐
specific manner, suggesting that the hippocampus is a mediator for sex dependent stress response. For further study more molecular analysis to elucidate the mechanism of GR will be done. 112.2 Brianna Harris Kyle Frantz, PhD Microglia activation and monocyte chemotaxis in RGS10-­‐null mice B. HARRIS, G. KANNARKAT, M. TANSEY Spelman College Neurodegenerative disorders are marked by peripheral immune cell infiltration into the central nervous system (CNS). On entering the diseased CNS, monocytes differentiate into tissue-­‐resident dendritic cells and macrophages which aid in modulating inflammation, producing effector molecules that destroy pathogens, and even help in brain-­‐resident microglial cells. Monocytes are brought to inflammation sites including the CNS through chemokine receptors (CRs). CRs are G-­‐protein linked receptors and are modulated by Regulator G-­‐protein Signaling (RGS) proteins, which act as GTPase accelerating proteins (GAPs). Previous studies have not identified physiologic substrates for RGS10, a GAP that is highly enriched in immune cells, implicated in regulating activation of microglia, CNS-­‐resident myeloid cells, and linked to age related macular degeneration in humans. The current study proposes to define the role of RGS10 in monocyte responses to chemokine stimulation. Chemotactic responses have been modeled by 78 seeding human umbilical vein endothelial cells (HUVECs) into a microfluidics chamber to measure binding patterns of monocytes from RGS10 wild-­‐type and knockout mice. This study also sought to confirm the microgliosis phenotype found in RGS10 knockout mice on a pure genetic background, the C57/BL6 background. New information obtained in this study could eventually lead to the development of treatments of neurologic diseases. 113 Matthew Phillips Linda Rinaman The effects of chronic food restriction on behavioral and hindbrain noradrenergic/peptidergic neural responses to cognitive stress M.B. PHILLIPS, H. ZHENG, L. RINAMAN Furman University, University of Pittsburgh Food intake and cognitive stress have been shown to independently activate two distinct but intermingled populations of neuronal cell bodies located in the caudal nucleus of the solitary tract (cNST) of the rat hindbrain. The first population is a subset of noradrenergic (NA) neurons that also express Prolactin-­‐
releasing Peptide (PrRP); the second a grouping of non-­‐adrenergic cells that express Glucagon-­‐like Peptide 1 (GLP-­‐1). These neural populations project directly to the bed nucleus of the stria terminalis (BST) and paraventricular nucleus of the hypothalamus (PVN) and are considered to play key roles in driving coinciding endocrine and behavioral functions related to regulating food intake, such as satiety, satiation, and stress-­‐induced hypophagia. Previous studies have shown that while these neurons can be activated independently by feeding and external stressors, disruption of feeding via overnight food deprivation strongly attenuates the responses of these hindbrain neurons to stress. To further delineate the relationship between food intake and stress response in context of this circuitry, we tested the effects of a novel feeding paradigm, chronic food restriction (CFR), upon rat performance in elevated plus maze (EPMZ), open field (OF), and social open field tasks. Brain tissues were also immunocytochemically double-­‐labeled for cFos and GLP-­‐1 or triple-­‐labeled for cFos and PrRP/dopamine-­‐β-­‐hydroxylase (DBH) to localize activation levels of cNST, BST, and PVN neurons in response to physical restraint. It was expected that rats with a greater degree of food restriction (70% of average maximum daily intact, CFR70) would show abolished GLP-­‐1 and PrRP/DBH activity in comparison to the group receiving 100% of their average maximum daily intact (CFR100), and would display greater degrees of exploration in the behavioral assays. Although histological analysis has not yet been completed, behavioral results have been promising. CFR70 possessed significantly higher open arm/closed arm and center zone/outer zone duration ratios than CFR100 in EPMZ and OF tests respectively without differing in mean distance traveled, suggesting greater explorative activity without increased locomotion. These behavioral results imply a strong anxiolytic CFR effect, further supporting the view that food intake and stress are intertwined via shared neural circuitry. Supported by the Furman Advantage Grant and the Rinaman Lab of the Department of Neuroscience, University of Pittsburgh Email: matt.phillips2892@furman.edu 114 Jayson Cruz Dr. Elizabeth Ann Becker Paternal retrievals increase testosterone levels in both male and female offspring in California mice (Peromyscus californicus). J.P. CRUZ, M.C. CHARY, M. BARDI, E.A. BECKER Saint Joseph's University The importance of maternal care effects on offspring development has received considerable attention, although more recently, researchers have begun to focus on the significance of paternal contributions. In the monogamous and bi-­‐parental California mouse (CA), fathers provide high levels of care and therefore 79 are a model system for studying paternal effects on behavior and their underlying neuroendocrine mechanisms. Likely through modulation of the arginine vasopressin (AVP) system which is androgen dependent during development, retrieval behavior by CA mouse fathers influences the development of aggression and parenting in adult male offspring. Male offspring exposed to paternal retrievals experience a transient increase in testosterone (T) which may underlie these observed changes in offspring brain and behavior. Few studies have examined father-­‐daughter interactions, although paternal retrievals have been shown to increase aggression in female offspring. Although T has been implicated in the regulation of female aggression, it remains unclear whether T may underlie changes in female offspring aggression in response to paternal retrievals. In the current study, we examined the influence of paternal retrievals on T in both male and female offspring. Retrievals were manipulated experimentally by displacement of the pup. Trunk blood was collected from retrieved, non-­‐retrieved and non-­‐manipulated (control) pups. We found that fathers expressed similar levels of retrievals towards sons and daughters, and that T levels were elevated in retrieved, as compared to non-­‐retrieved offspring. Similar to what has been previously described in male offspring and replicated here, female offspring that were retrieved had higher T levels than non-­‐retrieved females. Our data therefore suggest that paternal retrievals may serve similar functions in male and female pups i.e., fathers may shape adult offspring behavior via changes in circulating hormones. Email: jc564817@sju.edu 115 Jennifer Christensen Mary Harrington Foul weather friends: the role of oxytocin in meadow vole seasonal sociality. J.D. CHRISTENSEN, A.M.J. ANACKER, A.K. BEERY Smith College As research in the field of neuroendocrinology has gained momentum, the neuropeptide oxytocin has come into focus. Recent findings indicate it has influence on several aspects of social behavior. Voles (Microtus spp) have become a valuable model in social behavior research given the variation of social structures across species. Our research questions focus the effect of oxytocin on the affiliative bonds formed in winter months between female meadow voles (M. pennsylvanicus). While the affiliative behavior can be elicited in laboratory settings by simulating short daylight conditions, the question of neuronal mechanism remains. Female meadow voles express high levels of oxytocin receptors in the lateral septum; therefore, we infused oxytocin, oxytocin receptor antagonist (courtesy of Dr. Maurice Manning, University of Toledo, OH), or artificial cerebral spinal fluid into this region. Infusions were followed by twenty-­‐four hour cohabitation with a conspecific female. Social behavior of focal animals was then assessed in a partner-­‐preference test. Preliminary data indicate a trend toward lower levels of huddling when oxytocin or oxytocin receptor antagonist are administered. Lower levels of huddling with a partner following oxytocin infusion, relative to vehicle, demonstrate a role for oxytocin inhibiting social affiliation in female-­‐female bonds, contrary to findings showing a role for oxytocin promoting social affiliation in other brain regions in other species. A similar decrease in huddling following infusion of oxytocin with the oxytocin receptor antagonist also indicates that the action may be through the vasopressin receptor instead of the oxytocin receptor. Further aspects of our studies will address whether the vasopressin system may be involved, as well as the role of other high oxytocin receptor expression brain regions, which will assist in understanding the role of oxytocin in female meadow vole seasonal sociality. Email: jchristensen@smith.edu 80 116 Eyerusalem Lemma Sarah A. Heimovics The relationship between brain steroidogenesis and individual variation in singing behavior: a seasonal comparison E.M. LEMMA, K.M. CARLSON, S.A. HEIMOVICS University of St. Thomas European Starlings sing all year, however the motivation driving vocal behavior differs in breeding and non-­‐breeding contexts (Heimovics & Riters, 2007). Studies on brain regulation of vocal communication to date show support for seasonal context dependent differences in European Starlings, primarily in neurotransmitters such as dopamine (Heimovics & Riters, 2008) and opioids (Riters et al., 2005). It is unclear if steroid hormones, particularly steroids made in the brain, also have context dependent differences in the brain. The focus of this research was to investigate the context dependent differences in neurosteriodogenesis between nonbreeding and breeding season like endocrine states. Fourteen birds were randomly assigned to non breeding and breeding groups and underwent photoperiod manipulation to induce each endocrine state. Behavioral observations were performed in a free flying aviary for one hour a day for five consecutive days using a point sampling technique. Immediately after the last observation tissue was collected. Brains and gonads were flash frozen and stored in -­‐80ºC and gonad measurements were collected. Brains were sectioned at 200 microns and nuclei were punched and analyzed for gene expression using two-­‐step RT-­‐qPCR. Breeding condition birds had significantly larger gonads than non breeding birds showing that photoperiod manipulations were successful in inducing the endocrine states. Results show that there is not a significant difference in song rate between nonbreeding and breeding birds. In nucleus taenia of the amygdala (TnA) and and the bed nucleus of the stria terminals (BST), nuclei implicated to social behavior (Heimovics & Riters, 2005; Heimovics & Riters, 2006), individual singing behavior is positively correlated to individual mRNA expression of aromatase in breeding season birds, but not in nonbreeding season birds. In contrast, in the caudomedial nidopallium (NCM), an auditory processing region (Gentner et al., 2004), and rostral hypothalamus, a nuclei implicated in social behavior (Heimovics & Riters, 2005), no significant correlation exists in either context. This suggests that neuroestrogen production in TnA and BST is critical to sexually motivated vocal communication. Email: lemm8531@stthomas.edu 117 Bradley Wise Teresa Aubele-­‐Futch A nitric oxide promoter in the medial preoptic area facilitates copulation in adult male rats B. WISE, T. AUBELE-­‐FUTCH, E. HULL Wabash College Dopamine (DA) in the medial preoptic area (MPOA) is crucial in order for adult male rats to perform their entire range of copulatory behaviors. Previous research has shown that the nitric oxide (NO)-­‐cGMP pathway modulates and facilitates MPOA DA levels, and that administration of a NO synthesis inhibitor directly to the MPOA inhibits both basal and female-­‐stimulated MPOA DA release, stimulus sensitization, and copulatory ability. Thus, we hypothesized that administration of a NO promoter, sodium nitroprusside (SNP), could mimic the facilitative effects of stimulus sensitization on copulatory ability. In naïve male Long-­‐Evans Blue Spruce rats, SNP or saline was microinjected directly to the MPOA every other day for a total of 7 administrations. Other naïve male rats were exposed to receptive females placed over their cages on the same schedule, and a third cohort received IP injections of SNP on the same schedule. Those receiving SNP or saline did not receive exposure to females or female odors until copulatory testing. In a drug-­‐free test after the 7th administration of SNP, saline, or exposure to a receptive female, copulatory behaviors (mounts, intromissions, and ejaculations) were scored in a single copulatory session lasting thirty minutes from the first intromission. Animals that received SNP directly to 81 the MPOA showed facilitation of some, but not all, sexual behaviors, including ejaculation frequency and intromission-­‐to-­‐mount ratios over discrete ejaculatory series, similarly to those males that were stimulus-­‐
sensitized via exposure to receptive female rats. Animals receiving SNP regardless of the route of administration also showed lower post-­‐ejaculatory intervals, similar to female-­‐exposed animals. Thus, a NO promoter administered to the MPOA can mimic some facilitative aspects of stimulus sensitization in adult male rats. Email: bmwise14@wabash.edu 118.1 Sophia Stone Louise M. Freeman Sex Difference in the Medial Preoptic Area of the Asian Musk Shrew S.A. STONE, C.L. WRIGHT, M.M. MCCARTHY, L.M. FREEMAN Mary Baldwin College The medial preoptic area (mPOA) of the hypothalamus regulates sexual behaviors in many mammalian species. In rats, neural aromatization of testosterone to estradiol (E2) masculinizes both mounting behavior and dendritic spine density in the male mPOA by activation of COX-­‐2 and prostaglandin-­‐E2 (PG-­‐
E2); however, it is not known if this mechanism is shared by other species. Aromatization of androgens is not essential for sexual differentiation in the primate brain or the Asian musk shrew (Suncus murinus). Here, dendritic spine density in the shrew mPOA was evaluated via western blotting for spinophilin and direct quantification of dendritic spines in Golgi Cox-­‐impregnated neurons. Western blot analysis demonstrated that the male shrew mPOA contains more spinophilin than the female (p = 0.042), indicating a higher spine density in males, which we verified by quantifying spines visualized by Golgi-­‐Cox stain (p = 0.033). Further, we predict that treatment of female shrews with non-­‐aromatizable dihydrotestosterone (DHT), but not E2, would masculinize spinophilin levels along with mounting behavior. We hope to establish a biological as well as behavioral marker for brain masculinization in the musk shrew, and test the ability of COX-­‐2 and PG-­‐E2 manipulations to mimic the masculinizing effects of DHT. Email: stones8956@mbc.edu 118.2 Kaela Kelly Louise M. Freeman The effect of time of day on musk shrew sexual behavior K.J. KELLY, K.R. VASILOFF Mary Baldwin College Suncus murinus, also known as the Asian musk shrew, is a model species for studies of sexual behavior. The unique mating behaviors of musk shrews are due to the fact that the female is the aggressor. Initial mating practice begins with female lunges towards the male. Instead of the female performing lordosis, she instead tail wags to give indication of sexual receptiveness. The male will then proceed to mount the female until ejaculation. One of the advantages of using musk shrews includes that musk shrews do not have behavioral hormonal estrus cycles. Female musk shrews also only have a total gestation period of 30 days. Though this process has been observed in research, the optimal time for mating has not been studied. In this experiment, we looked at the musk shrews’ sexual behaviors and how they differed between morning and evening. Because previous laboratory matings have been performed in the evening, we expected to see more successful matings during that time. The results from this study can aid future matings to be performed at a time where breeding is more efficacious. To look at the effect of a multitude of variables on shrew sexual behavior, we observed a total of 14 female shrews and 14 male shrews. To control specifically for the differences of mating behavior we recorded time of day, humidity, successful mating counts, aggressive lunge counts [female aggression in the form of an audible shriek and physical approach], and whether or not the male ejaculated. The first observations were conducted as a baseline, 82 looking at normal behavior during the morning and evening. The second set of observations had a total n of 10, with an n of 5 in both the morning mating and evening mating groups. Two-­‐way chi square showed significance between ejaculation and morning or evening matings [X2(1)=4.16, p<.05]. There was no significant difference between the amount of lunge counts and time of day, nor was there any significant difference between humidity and ejaculation. This suggests that the optimal time to mate shrews may be at the beginning of their light cycle, rather than at the end as previously predicted. 119 Dana Cobb Mark Zrull Environmental enrichment during adolescence reduces affinity for novelty in young adult rats D. E. COBB, M. C. ZRULL Appalachian State University Like adolescents, young adult animals often exhibit increased novelty seeking, risk-­‐taking, and exploratory behavior. During adolescence,environmental enrichment (EE) can enhance brain development and learning and memory as well as affect variants of exploratory behavior including novelty seeking. In this study, we examined how EE during adolescence affected novel object and location preference in young adult rats and the relationship between these behaviors and neural activation in the basolateral amygdala (BLA) and dentate gyrus (DG). The BLA plays a role in processing emotion related to risk-­‐taking behaviors, and the DG contributes to modulation of these behaviors. Long-­‐Evans rats (n=16) were exposed to EE in cages with ramps, platforms, inanimate objects, and familiar and novel conspecifics for two days followed by a day without EE for a total of 20 EE sessions between postnatal days (PND) 34 and 64. Age-­‐matched control rats (n=16) were not enriched. Two-­‐trial object and location preference testing occurred between PND 66 and 75 (15, 30, 60 minute and 24 hour delays). On Trial 2, a novel object (NOP task) or familiar object at a novel location (NLP task) was present in the test field. Time in direct contact with and proximity to the novel object or at the novel location was measured. In the NOP task with 15 to 60 minute delays before Trial 2, the proportion of novel object contact time increased for control rats (+43%, 0.54 to 0.77) and dropped for EE rats (-­‐34%, 0.59 to 0.39). This trend reversed with a 24 hour delay (p<.05) when EE rats spent more time with the novel object (0.64 vs. 0.59). During Trial 2 of the NLP task, EE rats gradually decreased attention to the object at the novel location across delays (-­‐31%, 0.70 to 0.48). Control rats showed a consistent proportion of contact time with the newly located familiar object (0.60) across all delays (p<.05). On PND 78 following 2 hours in the quiet and dark, rats were sacrificed and brain tissue processed to count baseline active neuron levels using the c-­‐fos protein as an indicator. In BLA of EE brains, there was a 19% reduction in active neurons relative to controls, and a history of EE reduced activated neurons in the DG by 44% relative to unenriched controls. Behavioral data suggest enrichment during adolescence promotes short-­‐term (less than 1 day) adaptation to the introduction of a novel item into a familiar environment but longer-­‐term adaptation to rearrangement of familiar objects in a known environment in our young adult rats. Neural data suggest a history of enriching experiences produces an analogous change in brain structures contributing control to novelty preference behavior by reducing numbers of active neurons. Email: cobbde@email.appstate.edu 120 Janace Gifford G. Andrew Mickley Proposed considerations for analysis of prepulse inhibition J.J. GIFFORD, R.A. ZACHARIAS, C.P. TURNER Baldwin Wallace University 83 Prepulse inhibition (PPI) is a task that is typically employed to measure the acoustic startle response in animals. PPI is often determined using an equation that indicates the percent change of startle response when a prepulse (warning) is provided prior to the startle stimulus. Conversely, one may analyze prepulse trials without using a percent PPI equation. The purpose of this study was to determine whether more robust differences could be detected using a percent PPI equation versus raw startle response data. A data set was used to investigate alternate methods of analyzing PPI data. Within these data startle response to three different prepulse levels (5, 10, 15 dB above background) and pulse only stimuli were recorded. Results indicated that the method of analysis of prepulse trials leads to different conclusions. Specifically, fewer group differences were present when using the percent PPI equation compared to raw startle response data. Increased p-­‐values are also typically observed when using the formula. Thus, employing the percent PPI equation may not detect marginal differences that are significant when analyzing raw startle response data. The percent PPI equation utilizes the average of pulse only trials. However these data indicate startle response changes over a series of trials in response to pulse only startle stimuli. Therefore, using the average of pulse only trials in the percent PPI equation may also contribute to an inability to detect subtle group differences because the startle response to pulse only trials is changing. Researchers should also be aware that prepulse intensity significantly affects the startle response. In particular, the higher the prepulse level, the more the startle response is inhibited. Employing various prepulse intensities therefore may not be desirable unless the researchers intend to analyze startle behavior at each prepulse level independently. Taken together these findings suggest that the type of analysis and use of differing prepulse levels may impact results of PPI data. 121 Matthew Gerlach Michael Watt Effects of adolescent social defeat and isolation on adult anxiety and depression M.J. GERLACH, G.L. FORSTER, M.J. WATT Basic Biomedical Sciences, University of South Dakota Adolescent exposure to social stressors such as bullying is associated with later anxiety and depression disorders, many of which are characterized by deficits in cognitive processes mediated in part by prefrontal cortex (PFC) dopamine (DA) activity. We have shown that male rats exposed to repeated social defeat in adolescence exhibit decreased PFC DA activity as young adults, but display risk-­‐taking rather than anxiety-­‐like behavior. However, subjects in our previous studies were socially housed. Given that teen bullying victimization is often associated with social withdrawal, here we investigated whether social isolation concurrent with social victimization during adolescence would prove to be a contributing factor to the emergence of anxiety-­‐ and / or depressive-­‐like behavior in adulthood. Adolescent male rats (postnatal day [P]35) were exposed to socially aggressive adult males for five consecutive days. Age-­‐
matched controls received no social defeat, but instead were exposed to a novel empty cage each day. Half of all subjects were housed singly (isolated) starting at P35 and continuing to behavioral testing in early adulthood (P56–60). Anxiety-­‐like and depressive-­‐like behaviors were assessed using the elevated plus maze (EPM) and forced swim test (FST), respectively. No differences were found among groups in time spent in open arms of the EPM on the elevated plus maze. However, socially-­‐housed controls showed unexpectedly high anxiety-­‐like behavior, which may have obscured any effects of adolescent defeat and/or isolation. Therefore, we are repeating the EPM test with different subjects, and also assessing anxiety-­‐like behavior using the open field test. In the FST, socially isolated rats displayed less floating and increased swimming / climbing, regardless of exposure to adolescent defeat. This contrasts with effects of adult social isolation, which has been shown to enhance behavioral despair as indicated by increased floating in the FST. Similarly, previous studies indicate that social defeat in either adolescence or adulthood increases FST immobility. Contrary to our original hypothesis, our findings suggest that social isolation from mid-­‐adolescence to early adulthood may actually reduce depressive-­‐like behavior, mitigating negative effects of defeat exposure and increasing resilience to situations of inescapable stress. However, effects of combined social defeat and isolation in adolescence on later anxiety-­‐like behavior are yet to be resolved. Support: USD SPURA program (NIH NIDA grant R25-­‐DA033674) and NSF 1257679 (MJW). 84 Email: Matthew.Gerlach@coyotes.usd.edu 122.1 John Tishler Erin Wamsley Memory consolidation during a brief period of waking rest: an EEG study J.W. TISHLER, K.B. BROKAW, S.A. MANCEOR, K.B. HAMILTON, A. GAULDEN, E. PARR, E.J. WAMSLEY Furman University Introduction Numerous studies have shown that sleep helps to facilitate memory consolidation. However, emerging evidence suggests that waking rest can facilitate memory consolidation even in the absence of sleep. The present study examined the effects of eyes-­‐closed rest on memory consolidation across a short delay, using EEG to assess the brain correlates of resting memory consolidation. Methods Participants (n=26) listened to a short story, immediately recalled everything they could remember about the story, and then spent the next 15min either resting with eyes closed, or completing a distractor task (the computer game Snood). A delayed recall test was administered immediately following the 15min retention interval. All subjects completed both the quiet rest condition and distractor task condition, in counterbalanced order. Results Following exclusion of outliers, quiet rest led to greater improvement in memory compared to the active condition (p=0.005). Additionally, there was a strong positive correlation between change in story recall across the retention interval and slow oscillation power at all electrodes (r=0.69, p=0.002, at C4 electrode). Contrary to our original hypothesis, there was also a strong negative correlation between alpha power at central and frontal electrodes and change in recall (r=-­‐0.60, p=0.009, at F3 electrode). Conclusion This study supports the hypothesis that even a short period of rest can support memory under optimal conditions. Slow oscillations have been shown to enhance memory consolidation in sleep, and this study shows that these oscillations may also play a crucial role in memory consolidation during wake as well. Support This research was supported by a scientific bursaries award from the BIAL Foundation. Email: ward.tishler@furman.edu 122.2 Kelly Hamilton Erin Wamsley Impact of future relevance on dream content and sleep-­‐dependent memory consolidation K.B. HAMILTON, S. MANCEOR, K. BROKAW, Y. GRAVELINE, E. PARR, A. GAULDEN, J.W. TISHLER, E.J. WAMSLEY Furman University Background Previous studies have shown that sleep following the learning of new information is beneficial for memory consolidation of this information. Information that we know to be relevant in the near future may be preferentially processed during sleep, and may be preferentially reflected in our dreams. The current study aims to test the hypotheses that information relevant to an individual’s future is more likely to be incorporated into dream content, and these memories will also benefit more from sleep-­‐dependent memory consolidation. Methods Participants (n=57) trained on a Virtual Maze Task (VMT) and Motor Sequence Typing Task (MST). Immediately following training on the memory tasks, participants were given instructions about the later testing session. Those participants in the “Expected” condition were instructed they would be tested later on the memory tasks, and those in the “Unexpected” condition were instructed that they would fill out additional paperwork later. Participants then either had nine hours of sleep or nine hours of wake prior to delayed testing on both memory tasks. EEG data were acquired throughout sleep. Performance data were analyzed using a 2 (Expected vs Unexpected) x 2 (Sleep vs Wake) ANOVA. Results For the MST, performance improved significantly more across sleep than 85 across wakefulness (p=.05). The benefit of sleep was numerically greater in the “Expected” group than the “Unexpected” group (non-­‐significant). For the VMT, sleep also had a numerically greater effect on improvement when the test was expected (non-­‐significant). Data collection is ongoing. Conclusions Preliminary analysis of this ongoing study supports the hypothesis that sleep enhances memory consolidation more when newly learned information is relevant to a participants’ future. Previous studies have shown this effects for procedural memory, however, the present study is the first to test the effect of expectation on spatial memory consolidation during sleep. These results may give insight to a similar mechanism for procedural and spatial memory consolidation in the sleeping brain. Support This research was supported by a scientific bursaries award from the BIAL Foundation. Email: kelly.hamilton2835@furman.edu 123 Ashvini Pandian Joaquin Lugo Repeated oral doses of aniracetam does not alter anxiety, locomotion, or learning and memory in adult C57BL/6J mice A. PANDIAN, T.W. ELSTON, G.D. SMITH, A.J. HOLLEY, N. GAO, J.N. LUGO Baylor University There is a growing community of individuals self-­‐administering aniracetam, a nootropic, for its purported cognition enhancing effects. Aniracetam is believed to be therapeutically useful for enhancing cognition, alleviating anxiety, and treating various neurodegenerative conditions. Physiologically, aniracetam enhances both glutamatergic neurotransmission and long-­‐term potentiation. Previous studies of aniracetam demonstrate the cognition-­‐restoring effects of acute administration in different models of disease. No previous studies have explored the effects of aniracetam in healthy subjects. We investigated whether daily 50 mg/kg oral administration improves cognitive performance in naïve C57BL/6J mice by a variety of aspects of cognitive behavior. We measured spatial learning in the Morris water maze test; associative learning in the fear conditioning test; motor learning in the accelerating rotorod test; and odor discrimination. We also measured locomotion in the open field test; anxiety through the elevated plus maze test and by measuring time in the center of the open field test; repetitive behavior through marble burying. We detected no significant differences between the naive, placebo, and experimental groups across all measures. Despite several studies demonstrating efficacy in impaired subjects, our findings suggest that aniracetam does not alter behavior in normal healthy mice. This study is timely in light of the growing community of healthy humans self-­‐administering nootropic drugs. Email: Ashvini_Pandian@baylor.edu 124 Hensley Barnes Lori McGrew Assessment of working memory and anxiety in Danio rerio following treatment with pharmacological agents H. BARNES, L. SHEPPARD, K. PARKER, I. WAHID, J. WESLEY, C. WYATT Belmont University Danio rerio, more commonly known as zebrafish, are an NIH model organism that has long been used for toxicology and development studies. More recently, Danios have been used for behavioral experiments including assessments of anxiety and working memory. Our lab utilized a T-­‐maze to measure working memory following treatment of the fish with several different pharmacological agents including triclosan, acetaminophen, and nicotine. We also used a diving tank to evaluate the effects of these agents on anxiety in the zebrafish. We found a correlation between increased anxiety and decreased working memory in the Danio's. We went on to treat the fish with agents designed to decrease anxiety (anandamide receptor agonist and buproprion) to see whether these treatments would improve working memory in the fish. 86 Finally, we tested a pre-­‐workout supplement that for its ability to increase anxiety in the fish. Overall, treatments that increased anxiety as measured using the dive tank paradigm had a negative impact on working memory as assessed in the T-­‐maze in Danio rerio. Email: hensley.barnes@pop.belmont.edu 125 Lila Tibbets Dwight Krehbiel Emotion effects on performance in a 3-­‐back task L.E. TIBBETS, A. GONZALEZ, G.D.S. GONZALEZ Bethel College, KS This study investigated effects of emotional facial expressions on performance in a working memory task. Working memory performance was tested using a 3-­‐back matching test with fear and neutral faces. Our design and concept were based on a similar study that used valenced words (Kopf, Dresler, et al., 2013). Faces were selected from the Karolinska Directed Emotional Faces database (Lundqvist, D., Flykt, A., & Ohman, A., 1998). The 3-­‐back task consisted of two categories of trials -­‐-­‐ target faces (30 fear and 30 neutral), which had been seen three trials previously, and non-­‐target faces (66 fear and 66 neutral), which had not. The proportion of correct answers for target and non-­‐target faces and median reaction times for both conditions were measured for comparison. Measures for fear and neutral faces were obtained separately. EEG data were simultaneously recorded, and event-­‐related potentials (ERPs) were extracted. Participants were 20 Bethel College undergraduate student-­‐athletes. Two-­‐way, within-­‐subjects ANOVAs were used to determine possible effects of emotion and target/non-­‐target on proportion of correct answers and reaction times. No significant effect of emotion was found on either variable, nor was there an interaction between emotion and target/non-­‐target. However, the proportion correct for non-­‐target faces was significantly higher than that for the targets (Non-­‐Target M = .911, Target M = .780), indicating that non-­‐target faces were responded to more accurately than target faces. Box-­‐Cox transformations were used to achieve approximately normal distributions prior to conducting the ANOVAs. ERP results showed an enhanced late positive potential (LPP) for target faces, especially for fear target faces, compared to non-­‐
target faces at electrode sites Cz, Pz, Cp1, and Cp2. ERP results support the finding regarding processing differences between target and non-­‐target faces; enhanced by fear expressions in targets. ERP results were similar to those of Kopf et al. (2013), who found an enhanced LPP for negative valenced stimuli in the ERP average for electrodes Cz, Pz, CPz (not recorded in our study), CP1 and CP2. In addition, our ERP results showed an enhanced LPP for fear target stimuli at PO3 and PO4. Thus, both studies show an enhanced LPP for negative valenced stimuli. In comparison to the Kopf et al. study, we did not find a significant effect of emotion on behavioral variables; we found an enhanced LPP for target faces compared to non-­‐target faces, especially for fear. Thus, there is some doubt about whether perception of emotion in faces has similar effects as do words. Further research is needed to determine the source of these possible differences. Email: lilaetibbets@bethelks.edu 126.1 Mary Pearce J. Matthew Kittelberger Mapping immediate early gene activity in the brains of naturally vocalizing midshipman fish (Porichthys notatus) M.E. PEARCE, J.M. KITTELBERGER Gettysburg College Midshipman fish (Porichthys notatus), a highly vocal species of toadfish, have become a model system for studying the structure, function, and evolution of vertebrate vocal-­‐motor systems. Territorial males nest in intertidal zones where they hum nocturnally to attract females, and grunt or growl to defend their nests 87 from intruders. The vocal circuit has been mapped and areas involved in vocalization have been identified by a variety of lesion, chemical inactivation, stimulation, and a few single unit recording studies, as well as anatomical tract-­‐tracing experiments. Historically, functional studies of the vocal-­‐motor system have been performed in a fictive vocal preparation, in which vocal activity is elicited by electrical stimulation and monitored by electrodes placed on the motor nerve innervating the swim bladder (the vocal organ). No experiments have yet been done confirming that brain areas identified by these methods are indeed active in naturally humming fish, or if there are other important vocal areas active in natural humming but not in the fictive vocal context. Here, we used an antibody against the phosphorylated form of the ribosomal S6 protein, recently characterized as a downstream target of immediate early genes classically used to identify active neurons, to label neurons in naturally humming and non-­‐humming territorial male midshipman. Fish were day-­‐night reversed and monitored during dark hours using a hydrophone in tanks with artificial nests and females to induce natural courtship behavior. Males classified as humming were actively humming for at least one hour prior to sacrifice, and had inflated swim bladders. Non-­‐humming males were observed to be quiet, but listening to humming of other males in the same tank, for one hour prior to sacrifice, and were confirmed as non-­‐hummers by their deflated swim bladders. Brains of humming males were sectioned and stained with standard immunohistochemical methods for the expression of pS6. Labeling was cataloged, with attention paid to areas of the brain previously shown to be part of the vocal-­‐motor pathway. Robust staining was observed in the hindbrain vocal pattern generator in humming male midshipmen, in the vocal motor, vocal pacemaker, and pre-­‐pacemaker nuclei. However, no structures previously identified as vocal areas in the midbrain, hypothalamus, preoptic area, or forebrain were labeled. These results were based on a small number of fish, and further experiments will be performed to replicate the results and compare to non-­‐humming fish. Supported by the Grass Foundation and by a Howard Hughes Medical Institute Undergraduate Science Education grant to Gettysburg College. Email: pearma01@gettysburg.edu 126.2 Alexandra Turano J. Matthew Kittelberger The effect of early adolescent play behavior in rats on late adolescent and adulthood alcohol consumption A. TURANO, C.C. GARLISS, E.J. ACHTERBERG, J.G. LOZEMAN-­‐VAN 'T KLOOSTER, A.M. BAARS, L.J.M.J. VANDERSCHUREN, S.M. SIVIY, H.M.B. LESSCHER Gettysburg College; Utrecht University Play behavior during early adolescence has a strong correlation to social behavior later in life. Access to play during adolescence can increase social ability in late adolescence and adulthood, while denied access to play during adolescence can hinder future social capabilities. Individuals with impaired social aptitude are at a higher risk for self-­‐isolation and/or substance addiction. There is a high degree of individual variation in the risk for addiction. There is a possibility that individual variation in play behavior exhibited during adolescence is predictive of the risk for substance addiction habits in the future. The purpose of the current study was to determine if play behavior of early adolescent rats was related to late adolescent and adulthood alcohol consumption. 48 Lister-­‐Hooded rats were housed socially upon arrival and isolated for two and a half hours prior to play testing on pnd 28 and pnd 35, when play behavior is highly abundant. Play pairs were determined by weight and novelty to each other; individual rats were coded for pins and pounces. Subsequently, a ranking score for the number of pins and pounces in the two play sessions was calculated for each individual rat to differentiate high and low playing rats. Between pnd 42 and 56, half of the rats were given a free choice between water and 20% ethanol in a home cage intermittent-­‐every-­‐day paradigm, 7 hours a day on Monday, Wednesday and Friday for a total of 6 days. The other rats consumed merely water. A tertile split for play behavior indicated that high playing animals made significantly more pounces and pins. Moreover, the high playing animals consumed more alcohol when compared to low playing animals. Ongoing experiments with these animals will assess the relation between early adolescent play behavior and alcohol consumption in adulthood. Supported by NIMH grant R15MH100585 to S.M.Siviy 88 Email: turaal02@gettysburg.edu 127 Leigh Andrews enyhus@bowdoin.edu Selective attention and memory: Event related potentials and the IOR effect. L.A. ANDREWS, J.M. MACDONALD, J. MARKANT, E. NYHUS Bowdoin College Visual learning is dependent on the focusing of the attentional system. Studies show that attentional enhancement and inhibition modify the encoding of otherwise identically presented stimuli. fMRI has shown that activation for the attended stimulus was stronger with simultaneous inhibition at the distractor location utilizing inhibition of return (IOR), in which an elongated cue to target interval inhibits attention at the cued location and enhances attention at the non-­‐cued location. EEG research has linked the P1 component to attentional inhibition that occurs early in IOR while the Nd250 component is related to excitatory processes that override IOR. It has also been proposed that the FN400 component indexes familiarity and the parietal old/new effect indexes recollection during retrieval. The present study utilized IOR to study the impact of target enhancement and distractor inhibition on memory encoding. In the task a cue appeared on the left or right 600 ms before the stimulus appeared, subjects then responded to the target location. Participants were not informed that the images used would be the subject of a subsequent recognition memory test. EEG was used to look for component differences between target enhancement and distractor inhibition and between recognition and familiarity. Subjects’ RTs were faster for non-­‐cued than cued targets showing an IOR effect. Importantly, subjects’ memory was better for non-­‐cued than cued targets. In addition, preliminary results suggest an increased P1 and Nd250 for non-­‐cued relative to cued targets. These results indicate that subjects’ memory is affected by the selective attention during encoding. Paller Neuroscience Fellowship Loewy Neuroscience Fellowship Bowdoin College Neuroscience Department. Email: landrews@bowdoin.edu 128 Michele Amato Jeffrey L. Calton NMDA blockade and disorientation disrupts the establishment of a stable head direction signal M.E. AMATO, A.M. RODRIGUEZ, L.E. BERKOWITZ, I. YBARRA, J.A. JONES, I.A. PASTOR, J.L. CALTON California State University, Sacramento Head direction (HD) cells, found in many areas of the rodent Papez circuit, are thought to reflect the spatial orientation of the animal. Each HD cell fires maximally when the head is oriented towards a particular direction, known as the preferred direction of that cell. The preferred direction of the cell is typically different between environments, and this preference is determined during the initial exposure to that environment. Given the known role of NMDA receptor-­‐mediated glutamatergic transmission in many forms of neuroplasticity, we sought to determine whether NMDA transmission is necessary for the HD cell to maintain directional specificity when encountering a new environment. Anterior thalamic HD cells were recorded after the animals were administered the competitive NMDA antagonist CPP (RS-­‐3-­‐2-­‐
carboxypiperazin-­‐4-­‐yl-­‐propyl-­‐1-­‐phosphonic acid; 10 mg/kg) or isotonic saline. In the first session following injection, the cells were recorded in a familiar cylindrical environment to determine the baseline directional activity of the HD cell. Then, the animals were exposed to a new environment through one of two methods. In the Saline/Walks and CPP/Walks conditions, a door was opened and the animals were allowed to walk from the familiar cylinder to a novel square environment. In the Saline/Manual and CPP/Manual conditions, the animals were removed from the cylinder, placed in a cardboard box, and 89 disoriented before being manually placed in the novel square environment. The cells were then recorded in the novel square environment to determine if there were differences in the quality of the directional signal in the new environment. In general there was an attenuation of the directional signal between the familiar and novel environments, providing evidence that the signal suffers some degradation during the initial exposure to a new environment. This attenuation was larger in the CPP conditions, suggesting that NMDA transmission may be involved in maintaining the directional signal in a new environment. Finally, the attenuation was greatest when the drugged animals were disoriented prior to being placed in the novel square, suggesting that the effects of NMDA blockade are greatest when the animal has no stable directional reference to carry into the new environment. In accordance with this observation, several cells in the CPP/Manual condition showed slow drifts in preferred direction during the sessions, an effect not observed in the other conditions. These results suggest that NMDA blockade interrupts the maintenance of the directional signal carried by the HD cell network when the animal is exposed to a new environment. This research was supported by NIH grant 1R15NS071470-­‐01. Email: mea97@csus.edu 129.1 Lana McDowell Stephen Siviy Effects of risperidone treatment on 5-­‐HT2a and DRD2 mRNA expression in F344 and Sprague-­‐Dawley rats L.S. MCDOWELL, N. LI, K.A. LIPSETT, S.M. SIVIY Gettysburg College Imbalances in dopamine and serotonin functioning appear to be underlying problems in several neuropsychiatric disorders, including schizophrenia and autism. Rats have been used as an effective model organism due to their exhibition of complex social behavior, particularly play. F344 rats are an inbred rat strain that exhibit less play behavior than other rats and are known to have impaired dopamine and possibly impaired serotonin modulation. In an attempt to elucidate the neurological differences between F344 rats and an outbred strain commonly used in behavioral research, such as Sprague Dawley rats, real time quantitative PCR was performed on homogenized frontal cortex RNA samples from F344 and Sprague Dawley rats that had been treated with Risperidone, a 5-­‐HT2A/D2 antagonist, or vehicle for two weeks. Risperidone is a common atypical antipsychotic that is also used for the treatment of autism. It was hypothesized that blocking these receptors will result in an increase in the expression of the mRNA for these receptors. The target genes were DRD2 and 5-­‐HT2A, while PPIA and YWhaz were used as reference genes. Preliminary data suggests that vehicle-­‐treated F344 rats express less mRNA for 5HT2A and D2 receptors than vehicle-­‐treated Sprague Dawley rats, but that risperidone-­‐treated F344 rats have increased expression of both receptors, possibly greater than that of untreated Sprague Dawley. If maintained after further data collection, these differences may reveal systematic variability in serotonin and dopamine receptors between these strains, which then may account for some of the observed behavioral variability. Support from HHMI grant to Gettysburg College Email: mcdola03@gettysburg.edu 129.2 Samantha Eck Stephen Siviy Early experience, oxytocin, and dysfunctional play in the F344 rat S.R. ECK, J. SOROKA, L.S. MCDOWELL, S.M. SIVIY Gettysburg College Previous work from our laboratory has shown that the inbred Fischer 344 (F344) rat is consistently less playful than other inbred and outbred strains. In order to determine the extent to which these strain differences can be accounted for by strain differences in maternal care, a cross-­‐fostering study was done 90 with F344 and Lewis rats. Entire litters were either cross-­‐fostered or in-­‐fostered and maternal care was quantified using a timed-­‐sampling procedure over 10 days starting on post-­‐natal day 2. F344 mothers spent significantly less time licking and grooming their pups than Lewis mothers, although there were no strain differences in the amount of time spent in arch-­‐back nursing. After weaning at 21 days of age, rats were tested for play with a standard Sprague-­‐Dawley play partner. Rats were tested after both 4 and 24 hours of isolation, with F344 rats found to be less playful overall than Lewis rats in terms of both nape contacts and complete rotations. Rearing did have an impact on play but only in Lewis rats. In particular, cross-­‐fostered Lewis rats were as playful after 4 hours of isolation as they were after 24 hours of isolation. Cross-­‐fostered Lewis rats were also more likely to respond to a nape contact with a complete rotation than in-­‐fostered Lewis rats. To assess whether strain differences in trait anxiety could account for any differences in play, anxiety was assessed through activity in an elevated plus maze (EPM) and novel open field. When tested in the EPM, F344 rats spent more time in the open arms than Lewis rats and this measure of anxiety was not affected by rearing condition. When tested in a novel open field, F344 rats spent more time in the center than did Lewis rats. However, cross-­‐fostered Lewis rats spent more time in the center than in-­‐fostered Lewis rats. These data suggest that, contrary to our original working hypothesis, juvenile F344 rats are less anxious than same-­‐aged Lewis rats. As a result, these data suggest that the dysfunctional play of the F344 rat is not a consequence of enhanced anxiety in this strain. While early postnatal experiences may be having an impact on Lewis rats, maternal factors do not appear to contribute significantly to the dysfunctional play of the F344 rat. To assess if differences in oxytocin functioning may account for these strain differences, an additional group of male and female F344, Lewis, and SD rats were used to quantify oxytocin (OT)-­‐positive neurons in the hypothalamus. While still preliminary, no discernible differences in the number of hypothalamic OT-­‐positive neurons were observed between the strains. Support from NIMH grant R15MH100585 to S.M.S. and HHMI grant to Gettysburg College Email: ecksa01@gettysburg.edu 130 John Mootz Sharon Furtak Perirhinal cortex involvement in fear conditioning to a discontinuous light stimulus J. MOOTZ, C. CALUB, A. BECKNER, S.C. FURTAK California State University, Sacramento Previous studies have shown that lesions to the perirhinal cortex (PER) impair fear conditioning to complex auditory cues, such as pre-­‐recorded rat ultrasonic vocalizations (Linquist, Jarred, & Brown, 2004). In particular, the discontinuous nature of ultrasonic vocalizations was found to depend upon PER processing during fear conditioning to these stimuli (Kholodar-­‐Smith, Allen, & Brown, 2008). The current experiment examined whether PER is lesions induced similar impairments in fear conditioning to a discontinuous visual conditioned stimulus (CS). In this study, Sprague-­‐Dawley derived Albino male rats were broken into two groups: Lesion and Sham. The Lesion group underwent surgery and received intracranial injections of NMDA to produce bilateral excitotoxic lesions targeted at PER, while the Sham group underwent a similar surgical procedure with the exception that no injection was made. Following recovery, all subjects were trained on a three-­‐day fear conditioning paradigm. Day 1, Fear Acquisition, consisted of 5 presentations of the CS (discontinuous light) paired with an unconditioned stimulus (US), a foot shock. The subsequent two days consisted of a Context Test, the rat was placed back into the conditioning chamber with no CS or US presentations, and a Light Test, the rat was placed into a new chamber (a context shift) and presented with the light CS with no US presentations for 6 mins. The order of tests was counterbalanced across rats in each group. Freezing behavior (no movement except that necessary for breathing) was monitored and recorded throughout the experiment. Results show animals with PER lesions froze significantly less than Sham animals to the discontinuous light CS during the Light Test. Additional histological analysis will be necessary to eliminate the possibility that amygdala damage contributed to the observed deficit. These results further support PER in the processing of discontinuous stimuli and extend this hypothesis to include modalities outside of the audition, in particular stimuli within the visual modality. 91 Email: jrkm1.0@hotmail.com 131 Raquel Candal Greg Butcher Music therapy and its effect on Alzheimer's disease and dementia R. CANDAL, S. IRONS, A. ROSS, G. BUTCHER Centenary College of Louisiana and Thiel College Alzheimer’s is a devastating disease with symptoms ranging from memory loss to behavioral changes, all of which can be detrimental to a person’s quality of life. In some cases, behavior changes such as agitation have been shown to improve after participation in music therapy. However, our understanding concerning which type of music therapy is most effective is incomplete. Given this, we hypothesized that patients participating in music therapy sessions designed around a customized playlist would show greater improvements, both quantitatively and qualitatively, in behavioral symptoms than participants with a random playlist or a no-­‐music control. The following study was designed to investigate whether random music can improve quality of life or if familiar music, and its associated memories, holds the key to better outcomes. Participants that had scored a 19 or less on the Mini Mental State Examination (MMSE) were included in the study. The participants were randomly sorted into a customized playlist group, randomized playlist group, and a no-­‐music control group. The effects of the therapy were measured using a pre-­‐ and post-­‐test MMSE and Neuropsychiatric Inventory (NPI). Preliminary analysis indicates that a customized playlist significantly reduced agitation compared to the random playlist and the no-­‐
music control. If consistent, this finding could be used to develop more effective music therapy programs for people living with Alzheimer’s disease. https://www.youtube.com/edit?o=U&video_id=_SaXoH_VHuE Email: rcandal@my.centenary.edu 132 Nicole Comfort Jade Zee Linear integration for perceptual behavior in mouse primary auditory and visual cortex M.H. HISTED, N.T. COMFORT, R.T. OHMAN, A.R. PERILLO, J.H.R. MAUNSELL Dept. of Neurobiology, Harvard Medical School and Program in Behavioral Neuroscience, Northeastern University Many mammalian species can treat sensory information similarly whether it is distributed over time or concentrated over a short interval. Such linear integration means that the product of stimulus duration and amplitude predicts perceptual performance, a rule called Bloch’s Law. This phenomenon can be created by computations in the cerebral cortex, as we have shown by directly stimulating mouse visual cortex (Histed and Maunsell, 2014). Linear behavior of cortical circuits might have been unexpected because cortical neurons’ spike activity is a non-­‐linear function of their inputs. However, behavioral linearity can be explained by a decoder that applies a threshold rule to total population spike count. Because near-­‐perfect sensory integration is seen in many species and many sensory systems, it is likely that linear integration is a computation that many cortical circuits can perform. To determine if auditory cortex as well as visual cortex can use linear integration to guide behavior, we trained mice to perform an auditory task in which we determine the limits of their perception for pure tones of varying duration and amplitude embedded in noise. We find that mice show linear integration, following Bloch’s law, for audition: detection threshold is related to the product of duration and tone amplitude (duration: 2-­‐100ms, N=2 mice). We previously found, using direct stimulation of visual cortex, that very small changes in neuronal activity can be detected and reported by mice. These small changes mean that individual neurons do not carry enough information for animals to do the task. Instead, a population of neurons must be decoded to achieve the observed behavioral performance. We find that neurons in mouse primary 92 auditory cortex are also modulated only weakly by stimuli near detection threshold (median change in firing rate over interval before behavioral response: 0.5 spk/s; 75th percentile 1.7 spk/s; N=13 single units and 47 multiunits, recorded extracellularly in awake mice). Thus, despite the widely different types of sensory input processed in primary visual and auditory cortex, both areas can support linear integration. The cerebral cortex might have evolved to support linear integration as one important computational regime. Email: nicolecomfort3@gmail.com 133 Cassidy White Andrea Tracy The effect of high-­‐fat diet consumption on hippocampal dendritic spine density and spatial memory performance C.M. WHITE, A.L. TRACY Grinnell College Though an abundance of evidence now correlates diet-­‐induced obesity with deficiencies in learning and memory, much remains unknown about the physiological mechanisms underlying these effects. To gain more insight about how prolonged high-­‐fat diet (HFD) consumption alters function in the hippocampus, a brain region associated with spatial memory, this study investigates the effect of diet on hippocampal dendritic spine density. 16, male Long-­‐Evans rats consumed either standard chow (4% fat) or a HFD (40%) ad libitum for 16 weeks. All rats then learned to escape a Morris water maze onto a fixed platform. Rate of acquisition across training trials was equivalent across diet conditions. One week after the training period, a recall test was conducted with the platform removed from the maze. HFD animals were significantly slower to reach the platform's previous location (M = 41.78, SD = 25.93; M = 5.64, SD = 6.17; p = 0.005) and crossed that location significantly fewer times (M = 0.63, SD = 0.74; M = 3.50, SD = 1.07; p = 0.001) than standard chow animals. Brain tissue was then collected and Golgi stained to investigate neuronal morphology in the hippocampus. Results of spine density analysis revealed that HFD rats had significantly greater densities than their chow-­‐fed counterparts (M = 0.076, SD = 0.010; M = 0.054, SD = 0.011; p = 0.001). These results contrast previous research suggesting that increased spinal density is associated with better learning and memory performance. However, the data are consistent with spinal density changes expected to occur following increased exposure to leptin, an adipocyte-­‐derived hormone that is elevated with HFD consumption. Further research is needed to more completely understand the functional relationship between hippocampal dendritic spine density and the consolidation and recall of spatial memory. Email: whitecas@grinnell.edu 134 Emily Mortimer Cara M. Constance Determining the developmental stage of onset of behavioral circadian rhythms in Hyla versicolor (Gray treefrog) E.N. MORTIMER, C.E. LOYOLA, A.B. MYER, C.M. CONSTANCE Hiram College Circadian rhythms play a crucial role in the biological functions of many organisms on Earth. Internal clocks control basic cycles in an organism, including the rest/wake and feeding cycles. It is beneficial to study model organisms such as Xenopus laevis which possess similar clock genes as humans. However, it is also informative to study rhythms of animals in a natural setting. We hypothesized that tadpoles of the tree frog, Hyla versicolor, are active at a different time of day from the adults of the same species as a defense mechanism from cannibalistic tendencies and to avoid competition for similar resources. H. versicolor tadpoles procured from the field were monitored in constant darkness and temperature 93 controlled environment with unlimited access to food for five circadian periods with a video camera and software that measured movement. Of the 18 tadpoles tested, only one of the monitored tadpoles showed a circadian rhythm in behavior with peak behavior occurring in the nighttime. The data suggests that tadpoles do not have a stable circadian rhythm that contributes to their survival, and that they are consistently active throughout the day and night. Even though a stable circadian rhythm for tadpoles was not discovered, the conclusion reached is important because it gives rise to a further hypothesis that circadian rhythms emerge in a later developmental stage of H. versicolor as they mature into adult frogs. Further testing will be performed during metamorphosis to pinpoint when rhythmic behavior initiates in this species. Email: mortimeren@hiram.edu 135 Max Feinstein Ryan Lacy The effects of aerobic exercise on impulsive choice in a rodent model M.A. FEINSTEIN, R.T. LACY, J.C. STRICKLAND, M.A. SMITH Davidson College Impulsivity is a cardinal component in several psychiatric conditions resulting in maladaptive behaviors. Binge eating, attention deficit hyperactivity disorder, and substance-­‐use disorders all involve some component of impulsivity in their respective pathologies. Additionally, impulsivity can be influenced by a multitude of environmental conditions, and various procedures exist to objectively measure and quantify impulsivity in both human and animal subjects. We investigated the effects of aerobic exercise on impulsivity in a rodent model using a delay discounting paradigm. Delay discounting is the phenomenon where subjects will decrease their valuation of a reward as the time to receive that reward increases. For example, an impulsive choice observed from a delay discounting perspective would be selecting a bowl ice cream now rather than abstaining and enjoying better physical health later. To assess the effect of exercise on delay discounting, we placed rats in two cohorts, an exercising group that had ad libitum access to a running wheel, and a sedentary condition that was only allowed locomotion within their home cages (interior dimensions of 50 x 28 x 20 cm). Every day rats were placed in an operant chamber with two levers; one lever was programmed to deliver one 45 mg grain pellet immediately after each lever press, and the second lever was programmed to deliver three 45 mg grain pellets at increasingly delayed intervals. As the time required for the rat to wait for the larger, later reward (LLR) increased, we observed both groups would discount this option and choose the smaller, sooner reward (SSR). By logarithmically transforming the ratio of responses for the LLR and SSR and plotting those values as a function of delay (also a logarithmically transformed ratio of delay for LLR and delay for SSR), we were able to isolate two different behavioral mechanisms contributing to impulsivity. We found that exercising rats were less sensitive to the effect of delay than sedentary rats, indicating exercising rats were less likely to discount a reward as a function of delay. In contrast, we found that exercising rats were more likely to discount the value of an LLR due to the effect of the magnitude of the reward, meaning that sedentary rats valued the LLR more than the exercising rats, assuming an equal delay. In conclusion, we found a two opposing effects of exercise on impulsivity. Exercise decreased impulsive choice by reducing sensitivity to delay, but increased impulsive choice by reducing sensitivity to reward magnitude. Future investigations of these effects may involve analyzing the neuroadaptations resulting from exercise and their relevance to impulsivity. Funding provided by Davidson Research Initiative. Email: mafeinstein@davidson.edu 94 136 Caitlin John Margaret J. Gill Impact of differential rearing on set-­‐shifting ability C.A. JOHN, M.R. CROMWELL, M.J. GILL North Central College Rearing rats in enriched (EC) or impoverished (IC) conditions induces long term neurological, behavioral, and cognitive changes. EC rats exhibit improved cognitive abilities, such as increased responding for sucrose and visual stimuli, compared to IC rats. In contrast, it appears that EC rats have poorer reward discrimination compared to IC rats, when given the choice between a smaller-­‐sooner or larger-­‐later reward. To determine if these differences were due to cognitive deficits in EC and IC rats, the current study utilized an automated operant task, that is equivalent to the Wisconsin card sorting task often used in humans (Floresco, Block, & Tse, 2008). Rats arrived at 21 days of age and were reared in their respective conditions for 30 days prior to testing in standard two lever operant conditioning chambers. After rearing, rats began training on a FR1 schedule with sucrose pellets as rewards. Rats were tested for their side-­‐bias prior to learning their first “rule” involving visual cue discrimination. A cognitive shift was then quantified as rats were required to shift responding to the non-­‐biased lever in order to obtain a reward. During the visual cue task, IC rats displayed a greater number of correct choices and a fewer number of incorrect choices compared to EC rats. Similarly, in this same task, IC rats required fewer trials than EC rats to reach criterion. Since EC and IC differences were observed during the second session of visual cue discrimination, while not following a shift to the new rule, results suggest that EC and IC differences in reward discrimination may be due to learning deficits, rather than cognitive deficits impeding discrimination of the reward. Research supported by North Central College Summer Undergraduate Research Program 137 Monica Doring Larry Normansell Effects of FG7142 on isolation induced stress in Sprague-­‐Dawley and Wistar rats M. DORING, S. SIVIY Gettysburg College In the current study, we examined the effects of early social deprivation on the sensitivity of GABAA receptors in Wistar and Sprague-­‐Dawley rats. We examined the behavioral effects associated with the benzodiazepine inverse agonist N-­‐methyl-­‐β-­‐carboline-­‐3-­‐carboxamide (FG7142). The effects of β-­‐
carbolines are stress sensitive and can have enhanced behavioral effects when tested under conditions of chronic stress. In previous open field tests, isolation reared rats were more immobile, less active, and spent more time in the center of the field than the socially housed rats. Methods. Male Sprague-­‐Dawley and Wistar rats were housed either socially or in isolation for 2 weeks beginning at about 30 days old. Rats were tested in an open field for a 60 minute baseline period, injected with FG7142 (10 mg/kg IP), and then tested for an additional 60 minutes. Results. During baseline testing, rats housed in isolation were more active, reared more, and spent more time in the center of the arena than those housed socially. The effects of isolation were comparable between the 2 strains although Wistar rats were more active than SD rats overall and spent more time in the center. After treatment with FG7142, rats housed in isolation continued to be more active and reared more than those housed socially although this was only apparent in Wistar rats. This suggests that Wistar rats reared in isolation are particularly sensitive to the effects of FG7142. Conclusions. Wistar rats housed in isolation seem to be more sensitive to the effects of FG7142 than Sprague-­‐Dawley rats. The results support the idea that GABAA receptors are stress-­‐sensitive. The results show that there are strain differences in responses to stress, such as chronic isolation, which can alter sensitivity to drugs acting at the GABAA receptor complex. Funded by Jack Shand Summer Student Research Fellowship to M.D. Email: dorimo01@gettysburg.edu 95 138 Jason Freedman Michael Loose Identifying the relative influence of multiple prior events on predictions of a probabilistic future: an artificial neural network analysis J. FREEDMAN, A. AMLIE-­‐WOLF, R. WITTENBERG, O. SHORHAM, S.R. ARONSON, M.D. LOOSE Oberlin College Department of Neuroscience When predicting future events most individuals tend to match their rate of predicting an outcome to the probability of that outcome occurring. We used an artificial neural network trained via back-­‐propagation (ANN) to test the hypothesis that multiple types of information influence such predictions. The model used events of recent previous trials as inputs (14 input nodes) with two or five hidden nodes and two output nodes. The sign of the difference between the two output nodes determined the network’s guess. Training sets were constructed (N=22) from behavioral data collected for an event-­‐related potential study in which the probability ratio was 63:37 (Society for Neuroscience 2012, 101.22). Each training set contained a 50:50 ratio of high and low probability predictions while the test set retained the original choice ratio (range of high predictions: 54% -­‐ 82%). Ten networks, each initialized with random weight values, were trained for each of the 22 data sets. After training, the ANNs were more accurate compared to before training at predicting subjects’ choices with the training set (mean improvement: 14.9 %, SD = 5.0%, p < 0.001), as well as with the test set (mean improvement: 9.5%, SD = 11.2%, p < 0.001). We compared 3 metrics for rank ordering the contributions of the various inputs to accuracy of the networks. We tested a new metric that used the magnitude of output node difference values (as a measure of network confidence) and two established metrics, a lesion method and a connection weight strength algorithm. We examined four types of input patterns that could have influenced each decision: Patterns that defied the expected probability, behavioral streaks, accuracy of the previous guess, and speed of the previous guess. The strongest correlations between input pattern metrics and actual behavior were achieved with the synaptic weight algorithm (mean r = .83) and the confidence metric (mean r = .81). Both metrics were more highly correlated (p < 0.05) than the lesion metric (mean r = .49). Each input pattern was found to be important for a subset of individuals. The interactions of two or more patterns often were important to successful predictions by the ANN, and no one strategy was found to be strongly predictive for all participants. Our results suggest that for simple probabilistic predictions people vary their strategies depending upon recent events and their decisions are influenced simultaneously by multiple types of information. We infer that attempting to fit probabilistic decisions with a single strategy like the win-­‐stay lose-­‐shift or the expectation matching strategy will fail to capture significant components of the decision making process. Email: jason@freedman.com 139 Melissa Rose Adrienne Betz Behavioral and immune responses in chronic and acute unpredictable restraint stress M. ROSE, C. LITTLE, B. DALENA, C. WHITELOCK, A.J. BETZ Quinnipiac University Restraint stress has been shown to cause structural and functional changes in the hippocampus and prefrontal cortex. First, we characterized the behavioral effects of chronic unpredictable restraint stress in male Sprague-­‐Dawley rats. Chronic unpredictable restraint stress was induced over a 21-­‐day period. Two, 30-­‐minute sessions of restraint stress were performed at unpredictable times. We found that there were significant decreases in weight change in animals subjected to chronic unpredictable repeated restraint stress. Thirty minutes post the second session of unpredictable restraint stress, glucose levels were measured for 21 days. We found that there were no significant differences between the control and restraint group but glucose levels significantly decreased across the 21 day period. Spleen and adrenal weights were decreased in chronic unpredictable restraint stress exposed animals on day 21. Additionally, 96 there were exploratory differences in the elevated plus maze in animals exposed to chronic unpredictable restraint stress compared to control animals. Next, we performed an experiment with animals subjected to acute unpredictable restraint stress with two, 30 minute sessions. Animals were immediately sacrificed and single cell dissociations were performed with the hippocampus and prefrontal cortex. Immunomagnetic isolation of microglia were performed and confirmed with flow cytometry. Preliminary evidence suggests an elevation of microglia in both the hippocampus and prefrontal cortex brain regions; however, further research will be required to validate this. These findings suggest expression of these immune cells is increased with exposure to restraint stress. Email: melissa.rose@quinnipiac.edu 140.1 Emma Brockway Paul J Currie An investigation of the role of central ghrelin on appetite, metabolism, and stress activation E.T. BROCKWAY, C.B. MOCTEZUMA, E.C. AGAN, J.A. SELVA, P.J. CURRIE Reed College Ghrelin is a gut-­‐brain peptide secreted peripherally and active at ghrelin 1a receptors distributed throughout the rat central nervous system. In addition to stimulating eating, the peptide is known to elicit alterations in energy homeostasis, metabolism, and anxiogenesis. In the present report adult male Sprague Dawley rats were implanted with unilateral guide cannula aimed at the hypothalamic paraventricular or arcuate nuclei, the midbrain ventral tegmental area, or basolateral amygdala. Each region expresses the 1a receptor protein. Ghrelin was administered at doses ranging from 25-­‐800 pmol and was dissolved in sterile water vehicle in a volume of 0.2 ul. In addition to food intake, we examined the effect of the peptide on energy substrate utilization (respiratory quotient; VCO2/VO2), measured via indirect calorimetry, as well as the impact of ghrelin treatment on performance in the elevated plus maze paradigm. While ghrelin elicited eating after injection into all targeted brain regions, the peptide most robustly increased RQ after hypothalamic administration. Higher doses of the peptide increased avoidance of the open arms in the elevated plus maze, indicative of the induction of stress-­‐related or anxiety-­‐like behavior. If rats were allowed to eat after ghrelin treatment, and then tested in the EPM, no anxiety-­‐like behavior was observed. Overall our findings further delineate the role of forebrain and midbrain structures mediating the effects of ghrelin in the expression of appetitive and emotional behaviors. Supported by the Murdock Charitable Trust. Email: pcurrie@reed.edu 140.2 Cloe Moctezuma Paul J Currie Systemic and brain ghrelin signaling in ethanol reward C.B. MOCTEZUMA, E.C. AGAN, E.T. BROCKWAY, J.A. SELVA, P.J. CURRIE Reed College In the central nervous system mesotelencephalic neurons are implicated in the control of reward and reinforcement. Numerous neurotransmitter and peptides are synthesized or expressed by these neurons and act locally in the mediation of drug reward, including ethanol reinforcement. Recent research indicates that the gut-­‐brain hormone ghrelin plays an important role in this process. For example ghrelin microinjection directly into the midbrain ventral tegmental area increases operant responding for palatable food, an effect blocked by central dopamine depletion. In the present study we investigated the impact of ghrelin on ethanol intake in male Sprague Dawley rats. Animals were initially exposed to increasing concentrations of ethanol. Midway through this exposure period rats were implanted with chronic indwelling guide cannula aimed at the ventral tegmental area or the nucleus accumbens. After rats 97 had showed stabilized intake of 8% ethanol, they were then injected with either ghrelin or sterile water vehicle using a repeated measures design. A period of at least 3 non-­‐injections days separated various treatments. A separate group of rats received peripheral ghrelin injections. Intakes were assessed at 2 and 6 hrs postinjection. Our findings indicated that both systemic and central ghrelin administration elicited reliable increases in ethanol consumption. Overall this work is consistent with the role of mesotelencephalic ghrelin signaling in the mediation of ethanol reward. Supported by the Murdock Charitable Trust. Email: pcurrie@reed.edu 141 Maxwell Anderson Gary Muir Influence of visual cues on head direction (HD) cell firing following anesthesia M. ANDERSON, M. SEVERSON, A. THOMSON, G. MUIR St. Olaf College One class of neuron thought to play a role in navigation and spatial orientation are head direction (HD) cells. HD cells fire relative to a rat's directional heading, and this firing is thought be controlled by the animal's use of both idiothetic (internal) and allothetic (external) cues. Our study investigates the effects of manipulating a prominent visual (allothetic) cue in a cylindrical testing chamber on HD cell directionality following a period of unconsciousness. Four HD cells were recorded from the Anterior Dorsal Thalamic Nucleus (ADN) of animals before (baseline), during, and following a period of unconsciousness induced by Isoflurane gas anesthesia. All HD cells appeared to lose their directionality during the period of unconsciousness, firing randomly in all directions, but quickly regained their directionality upon a return to consciousness. If the visual cue was visible during recovery, all HD cell firing returned to its original (baseline) direction. Surprisingly, when the visual cue was removed, three of the cells still reverted back to their original firing direction on recovery from anesthesia. These results indicate that when available, visual information significantly controls the directional firing of HD cells. When the visual cue is removed, however, unknown allothetic cues, or a preserved idiothetic signal despite the apparent randomness of the HD cell firing during unconsciousness, could possibly explain how the HD cell returns to its original firing direction. The next step in our ongoing research is to determine whether there is another, uncontrolled external cue influencing this outcome; primarily, whether directionality is maintained when visual and auditory cues are diminished (e.g., using complete darkness and loud white noise). Email: andersmr@stolaf.edu 142 John Connell Shelly Dickinson The impact of caffeine on the motivational effects of alcohol in mice J. CONNELL, M. NORBY, P. VUE St. Olaf College Combined caffeine and alcohol consumption is a common practice during adolescence, a period of acute neurobiological vulnerability. It is important to understand the behavioral and neuropharmacological effects of these substances on the adolescent brain as early exposure may contribute to the development of alcohol abuse. The present study employed place conditioning (PC) and conditioned taste avoidance (CTA) procedures to assess the impact of caffeine on the rewarding and aversive effects of ethanol in adolescent and adult mice. The PC procedure produced an aversion to a 20 mg/kg dose of caffeine after four trials as well as a preference for a 2 g/kg dose of ethanol after eight trials in adolescent mice. Both effects were subject to extinction, with the ethanol preference extinguishing most rapidly. In addition, the CTA data showed that the presence of caffeine, when combined with either ethanol or saline, produced a stronger aversion in both age groups compared to the saline and ethanol. Although it is unclear whether 98 these results were produced by behavioral summation or pharmacological interactions, such data provides further insight into the combined effects of caffeine and alcohol. Email: connell@stolaf.edu 143 Jordan Buck Jessica Siegel Effects of early adolescent methamphetamine and nicotine exposure on behavior and cognition in adolescent mice J.M. BUCK, A. SCHULTHEIS, J.A. SIEGEL The University of the South The neurotoxic effects of methamphetamine (MA) can lead to deficits in behavior and cognition. The rising rates of adolescent MA use necessitate that we understand effects of MA exposure on the adolescent brain. Adolescents in treatment for MA abuse show higher levels of depression and suicide ideation compared to those being treated for other substances. Adolescents using MA also show high rates of nicotine use. Previous research has shown that nicotine can mediate the effects of MA in the brain. However, the interaction between MA and nicotine in the adolescent brain, and the effects of these two substances, has not been examined. This research assesses the effects of early adolescent MA and nicotine exposure on cognition and behavior in male C57BL/6J mice later in adolescence. The effects of early adolescent MA and nicotine exposure on behavior in the open field test, the novel object recognition test, the Porsolt forced swim test, the Morris water maze test, and MA conditioned place preference were examined. Mice exposed to MA or nicotine in early adolescence showed increased time spent in the center of the open field compared to mice exposed to saline or both MA and nicotine together. There were no effects of early adolescent MA and/or nicotine exposure in early adolescence in any other test, including the conditioned place preference test. These findings suggest that early adolescent exposure to MA or nicotine increases risk taking behavior and decreases anxiety, but concurrent exposure to both MA and nicotine reduces this effect. Current experiments examining dopamine transporter density and corticosterone levels are ongoing to better understand the mechanisms underlying these behavioral results. These findings contribute to a greater understanding of how MA and concurrent nicotine exposure alters behavior and cognition in an age group that has been relatively understudied. These studies supported by: James D. Kennedy III Faculty Fellowship from The University of the South. Email: buckjm0@sewanee.edu 144 Torrie Summers Brian Burrell Endocannabinoids/endovanilloids attenuate injury-­‐induced hyperalgesia but not mechanical allodynia T. SUMMERS, B. HANTEN, W. PETERSON, B. BURRELL University of South Dakota The endocannabinoid (eCB) system is thought to play a role in modulating nociceptive signaling at both the central and peripheral levels, thus making it a potential therapeutic target for modulating pain signaling. The analgesic effects of eCBs are thought to be due to depressing excitatory synaptic transmission within pain neural pathways. However, results from both clinical and laboratory-­‐based studies have found that eCBs can also contribute to nociceptive sensitization (e.g. mechanical hyperalgesia or allodynia) due to a depression of inhibitory synapses that leads to disinhibition of pain circuits (Pernia-­‐
Andrade et al., 2009). Using the medicinal leech as a model system, our lab has observed that while eCBs depress nociceptive synapses, they enhance non-­‐nociceptive synaptic transmission (Yuan & Burrell, 2010). Here, we examined the functional relevance of these synaptic effects by testing whether eCBs have similar bidirectional effects on behavioral responses to nociceptive vs. non-­‐nociceptive stimuli. Leeches 99 were injected with either the eCB 2-­‐arachidonoylglycerol (2-­‐AG; 75µM) or anandamide (AEA; 100µM) and then tested for nociceptive and non-­‐nociceptive stimulus response thresholds. Both AEA and 2-­‐AG significantly sensitized the animals to non-­‐nociceptive stimuli while simultaneously decreasing responses to nociceptive stimuli. Both the pro-­‐ and anti-­‐nociceptive effects of AEA and 2-­‐AG were blocked by co-­‐
injection of an inhibitor of the transient receptor potential vanilloid (TRPV) channel, which is thought to function as an eCB receptor. Further experiments were conducted to determine if eCBs have an effect on non-­‐nociceptive and nociceptive stimuli responses in an injured animal. Animals were given a crush injury or injection of LPS (1mg/mL) to their posterior sucker and their behavioral responses to nociceptive and non-­‐nociceptive stimuli was observed daily for 14 days. Animals received an injection of 2AG (75µM) on the third day after injury. The injured animals had a significantly increased response to non-­‐nociceptive (allodynia) and nociceptive (hyperalgesia) stimuli. Treatment with 2AG restored responses to nociceptive stimuli to pre-­‐injury latencies; however, 2AG treatment did not attenuate the sensitized response to non-­‐
nociceptive stimuli. These results indicate that eCB treatment may be effective in treating mechanical hyperalgesia, but may not be useful in treating mechanical allodynia. 145 Jenna Goldstein Eric Wiertelak Huang lian jie du wan: traditional chinese medicine formulation for pain treatment J. GOLDSTEIN, C.SEMLA, G. THAYER, E. WIERTELAK Macalester College TCM is more than 5,000 years old and has its roots in the ancient philosophy of Taoism. Whereas Western medicine tends to focus on maladies of particular organs or parts of the body, TCM focuses on the balance and imbalance of various patterns, and utilizes these patterns to diagnose and treat patients. Some of the most important principles are cold/heat, damp/dry, excess/deficiency, and yin/yang. Huang Lian Jie Du Wan is a TCM formula that focuses on repletion heat, heat toxin, and damp-­‐heat. It contains 4 primary herbs, huang lian, huang qin, huang bai, and zhi zi, as well as 3 supplementary herbs that are specific to the formulation used in this study, including chuan niu xi, shi gao, and dan pi. The different herbs in Huang Lian Jie Du Wan have individually demonstrated health benefits but have not been studied together as a formula. This study used 3 different pain assays – tail flick, hot plate, and formalin – to assess Huang Lian Jie Du Wan’s analgesic properties in a rat model. The rats received a 5mL oral decoction of the formulation via gavage 1 hour prior to testing and were compared to a control group that received water. There was no significant change in pain response for any of the assays, running contrary to the hypothesis that the formulation would reduce pain responses. This result was unexpected, as prior research indicated that the herbs had potentially analgesic effects individually, therefore it was reasonable to assume that they would maintain, if not increase their analgesic properties when combined in a formulation. The lack of significant results may be due to the fact that Huang Lian Jie Du Wan is generally administered over the course of several days, as opposed to a one-­‐time dosage, offering insight into areas for future study. 146 Kathryn Hathaway Darcy Burgund Processing distracting text: A Biopac EEG test case K.V. HATHAWAY, E.P. WIERTELAK Macalester College The present study aims to validate the use of the Biopac Systems, Inc. EEG technology in a laboratory setting where higher-­‐end brain imaging hardware is not available. To test for validity and reliability, a replication of a cognitive task from a 2003 ERP study by Phillips and Lesperance was initiated using Biopac's MP36 console and Acqknowledge data recording software. In the original study, participants 100 were presented with sentences with imbedded distracter words and were asked to read aloud the sentences while ignoring the distracters. Participants were then presented with a probe word and the N400 waveform was analyzed for probing for either sentence-­‐related words, distracter-­‐related words or unrelated words. The researchers found that the electronegativities for distracter and sentence-­‐related words was not as high as those seen for unrelated probe words, demonstrating that distracters were being cognitively processed along with the sentences themselves. The results of the present study showed a similar pattern of priming, but only for distracter-­‐related probe words and not for those that were sentence-­‐related. Future studies should re-­‐visit the Biopac hardware in order to determine if the results were due to the equipment itself or to experimental error. Email: khathawa@macalester.edu 147 Brett Campbell Julia E. Meyers-­‐Manor Lending a helping paw: GABAergic mechanisms in empathy and pro-­‐social behaviors in rats B.A. CAMPBELL, J.E. MEYERS-­‐MANOR, N.D. MATHEWS, & E.P. WIERTELAK Macalester College Non-­‐human animals, here specifically rats, are capable of both pro-­‐social behavior and responses implicating emotional contagion. The current studies set out to: 1) determine whether rats may demonstrate the ability to recognize a conspecific’s distressed state from a non-­‐distressed state, through patterns of behavior indicative of differential levels of accord with an empathetic response, and 2) Examine the role that GABAergic mechanisms may play in such responsivity. The activation of GABAergic mechanisms is highly correlated with relief from anxiety-­‐related symptoms in humans; the question here was whether alterations in GABAergic activity might affect the performance of empathy-­‐related activity in rats. Physiological symptoms of anxiety and distress originate in activation of brain areas associated with the limbic system. Administration of chlordiazepoxide, a prototypical GABAergic benzodiazepine agonist results in decreased levels of such responsivity. To examine whether activation of limbic system structures and GABAergic mechanisms are necessary for the emotional contagion involved in empathetically motivated behavior, in study 1, subjects from two groups of rats were placed in individual plexiglas restraining apparatuses; one habituated to the restrainer, one non-­‐habituated (and therefore distressed). A free-­‐roaming cagemate was then placed into the open-­‐field containing the restrainer, which offered the option of opening the restrainer and freeing their cage-­‐mate. Here, rats released the distressed cagemate at a greater rate on the first day of testing than those in the habituated, non-­‐
distressed group. In study 2, non-­‐habituated distressed group received either 2 mg/kg of chlordiazepoxide or vehicle to evaluate the impact of benzodiazepines on pro-­‐social behavior. Email: bcampbe2@macalester.edu 148 Simon Sangaard Eric Wiertelak Kappa-­‐Opioid receptor agonist analgesia and antianxiety: Effects of Salvinorin A in rats E.P. WIERTELAK, S.A. ANDERSON, JR., H.C. GEMRICH, T.M. NICHOLS-­‐MEADE, S. SANGAARD Macalester College Salvinorin A is the main active component of Salvia Divinorum, a plant indigenous to Oaxaca, Mexico. The subjective effects of S. Divinorum are described as being akin to lysergic acid diethylamide (LSD) and other classical hallucinogens that interact with the serotonin 5-­‐HT2A receptor subtype. However, Salvinorin A does not interact with this receptor (Listos et al., 2011). Therefore, Salvinorin A can be defined as a structurally unique, non-­‐nitrogenous, highly selective kappa opioid receptor (KOR) agonist (Roth et al, 2002). Results from various studies exemplify a range of effects similar to known KOR 101 agonists, although reports of analgesia from Salvinorin A have been somewhat varied and collectively limited (McCurdy et al., 2006). Braida et al. (2009) found no dose-­‐dependent effects of Salvinorin A in the Elevated Plus Maze, which tests for anxiety. Their results suggest that Salvinorin A could possess a slight anxiolytic effect. This present study expands upon previous work by testing the analgesic and anxiolytic effects of Salvinorin A on rats using the hot plate, tail-­‐flick, formalin and Elevated Plus Maze assays at three different dosages of Salvinorin A: 0.25 mg/kg, 0.50 mg/kg, and 0.75 mg/kg. Significance was found in open-­‐arm time between the control and 0.50 mg/kg and between 0.50 mg/kg and 0.75 mg/kg. Behaviorally, rats given the 0.50 mg/kg dosage exhibited exploratory activity, as opposed to the 0.75 mg/kg, which induced hypolocomotion and avoidance of the open-­‐arms. These results suggest that 0.50 mg/kg of Salvinorin A has a significant anxiolytic effect in rats. Studies of the potential analgesic effects are currently ongoing. In the future, Salvinorin A could serve as a template for non-­‐addictive opioids, provided dose-­‐dependent dysphoria and hallucinations are eliminated. More research needs to be done to understand the mechanisms behind Salvinorin A, in order to utilize its possible anxiolytic and analgesic therapeutic effects. Email: ssanggaa@macalester.edu 149 Josh Rogers Kenneth Renner Organic cation 3 antagonist infusion into the mediobasal hypothalamus inhibits female sexual behavior J.T. ROGERS, M.W. BUCHANAN, J. CHILUWAL, K.J. RENNER University of South Dakota The dorsomedial hypothalamus (DMH) is believed to modulate stress through the integration of autonomic and neuroendocrine responses that mediate appropriate changes in behavior. Delivery of stress hormone, corticosterone (CORT) into the mediobasal hypothalamus, which includes DMH and ventromedial hypothalamus, of rats primed with estradiol (E2) and progesterone (P) markedly increases extracellular serotonin (5-­‐HT). CORT may exert rapid effects on 5-­‐HT by blocking organic cation transporters (OCTs), a nonspecific class of membrane transporters that are highly expressed in the DMH. Since OCTs are believed to function as a clearance mechanism for monoamines, including serotonin, it is possible that stress-­‐induced increases in CORT may exert behavioral effects by enhancing and prolonging 5-­‐HT effects. Normetanephrine (NORMET), a metabolite of norepinephrine, also blocks OCT3. We hypothesized that CORT and NORMET, by interfering with OCT-­‐mediated serotonin clearance in the DMH, would increase 5-­‐HT; acutely suppress the expression of the lordosis reflex and increase the expression of anxiety-­‐like behavior. Anxiety was tested using the open field test and the elevated plus maze. Female sexual behavior was tested by evaluating the expression of lordosis in response to 10 mounts by a male. In all tests, ovariectomized females were primed with E2 (5 µg/0.1 mL V, 24 hr) and P (0.5 mg/0.1 mL V, 4-­‐6 hr) and treated with bilateral infusions of CORT (48 pg/0.5 µL), NORMET (45µg/0.5 µL) or vehicle into the DMH 10 min prior to behavioral evaluation. Surprisingly, preliminary results suggest that neither CORT nor NORMET infusions into the DMH affect anxiety-­‐like behaviors. Previous work indicates that restraint stress markedly increases DMH 5-­‐HT activity in females. It is possible that the EPM and open field tests were not sufficiently stressful to elicit a difference in the expression of anxiety-­‐like behavior. However, our results indicate that infusions of CORT and NORMET into the DMH reversibly suppress sexual behavior. The effect of the OCT3 antagonists in inhibiting sexual behavior is consistent with an earlier finding that CORT increases mediobasal hypothalamic 5-­‐HT in steroid-­‐primed females and that 5-­‐HT in the hypothalamus is inhibitory to lordosis. Support: NSF IOS 0921874 Email: Joshua.rogers@coyotes.usd.edu 150 Shaydel Engel Lee Baugh 102 The role of corticotrophin releasing factor in mediating dopamine and serotonin in the nucleus accumbens: a potential mechanism for how stress may impact a reward system S. ENGEL, J. SCHOLL, L. BAUGH, G. FORSTER University of South Dakota Stress-­‐induced relapse is one of the major reasons for drug relapse. There are no current treatments, because the mechanisms are not fully understood. Corticotropin-­‐ releasing factor (CRF) is a stress hormone that causes an increase in the release of the neurotransmitter serotonin in the reward-­‐seeking brain region, the nucleus accumbens (NAc). It is known that infusion of serotonin into the NAc leads to an increase in the reward-­‐related neurotransmitter dopamine in the NAc. High levels of dopamine are related to drug seeking behavior and increased cravings during withdrawal, which often lead to relapse. This study examined whether CRF-­‐induced serotonin results in increased dopamine levels in the NAc, testing whether a neural pathway activated by stress can then activate a neural pathway related to drug seeking and relapse. Monoamines were collected from a probe in the NAc of male rats. Samples collected from the probe were injected into a high performance liquid chromatogram system (HPLC) to measure dopamine and serotonin. After baseline levels for each were established, a vehicle, or CRF (500ng) and vehicle were infused via cannula in the serotonin cell-­‐body region, the dorsal raphe nucleus (dRN). When CRF was infused into the dRN, serotonin levels increased in the NAc for a short time (20 min), coinciding with a larger and more prolonged increase in NAc dopamine levels. These results suggest that stress-­‐related increases in serotonin within the NAc may increase dopamine in this region, although the exact mechanisms need to be explored in further studies. Overall, these findings suggest that blocking CRF receptors during withdrawal periods may help minimize cravings, making recovery more achievable. Supported by: NIH NIDA grant R25-­‐DA033674. Email: shaydie.engel@coyotes.usd.edu 151 Alexis Tarter Jennifer Roxanne Prichard The Invisible Variable: Sleepiness and suicidality in a large national college health survey A.N. TARTER, J.R. PRICHARD University of St. Thomas Introduction Insomnia is both comorbid for and a consequence of depression. Longitudinal studies in adults with PTSD and population studies with school aged children have demonstrated that sleep problems are also independent predictors of suicidal thoughts and attempts. More research is needed to evaluate the interactions between poor sleep and suicidality in emerging adults, a vulnerable population who is at high risk for both mood disorders and profoundly disturbed sleep. Our study evaluated the relationships between sleep difficulties and self-­‐harm behaviors and suicidal thoughts in a large national sample of college students. Methods Data from the Spring 2009 American College Health Association National College Health Assessment-­‐II were analyzed for trends in daytime sleepiness, suicidality (self-­‐
harm behaviors, suicidal thoughts, and suicide attempts in the last year) among undergraduate students (n = 72,966) with diagnosed insomnia, with probable undiagnosed insomnia, and without major sleep initiation and maintenance problems. Results Five percent of the total poulation had been dignosed with or treated for insomnia within the last year. Of these students, 71.3% had a comorbid anxiety disorder and 62.2% had a comorbid depression diagnosis. Students with diagnosed insomnia were at elevated risk for self-­‐harm behaviors [O.R. 3.02, C.I. 2.74-­‐3.34], suicidal ideation [O.R. 3.28, C.I. 2.99-­‐3.60] and suicide attempts within the last year [O.R. 5.89, C.I. 5.01-­‐6.02]. Another 8.3% of the population had probable undiagnosed insomnia (defined here as sleep iniation and maintence problems >1.5 s.d. from the mean; trouble falling asleep approximately 6 days/week and awakening too early 4 days/week.) These students were also at elevated risk for self harm behaviors, suidicidal ideation and suicide attempts [O.R. > 2.7, for all cases]. Conclusion College health professionals can provide more effective and targeted interventions for students by understanding the elevated risk factors for self-­‐harm, suicidal ideation, and suicidal attempts in those with insomnia. 103 Email: tart4718@stthomas.edu 152 Samantha Herdegen Robert Calin-­‐Jageman Characterization of the rapid transcriptional response to long-­‐term sensitization training in Aplysia californica S. HERDEGEN, G. HOLMES, A. CYRIAC, I.E. CALIN-­‐JAGEMAN, R.J. CALIN-­‐JAGEMAN Dominican University We used a custom-­‐designed microarray and quantitative PCR to characterize the rapid transcriptional response to long-­‐term sensitization training in the marine mollusk Aplysia californica. Aplysia were exposed to repeated noxious shocks to one side of the body, a procedure known to induce a long-­‐lasting, transcription-­‐dependent increase in reflex responsiveness that is restricted to the side of training. One hour after training, pleural ganglia from the trained and untrained sides of the body were harvested; these ganglia contain the sensory nociceptors which help mediate the expression of long-­‐term sensitization memory. Microarray analysis from 8 biological replicates suggests that long-­‐term sensitization training rapidly regulates at least 81 transcripts. We used qPCR to test a subset of these transcripts and found that 83% were confirmed in the same samples, and 86% of these were again confirmed in an independent sample. Thus, our new microarray design shows strong convergent and predictive validity for analyzing the transcriptional correlates of memory in Aplysia. Fully validated transcripts include some previously identified as regulated in this paradigm (ApC/EBP and ApEgr) but also include novel findings. Specifically, we show that long-­‐term sensitization training rapidly up-­‐regulates the expression of transcripts which may encode Aplysia homologs of a C/EBPg transcription factor, a glycine transporter (GlyT2), and a vacuolar-­‐protein-­‐sorting-­‐associated protein (VPS36). http://authors.elsevier.com/a/1PabZ3qNa9iMVH Email: herdsama@my.dom.edu 153.1 Haley Turner Deanna Buffalari The effect of social interaction on ethanol consumption H. TURNER, D. BUFFALARI Westminster College Alcohol use has been defined as a public health issue in the United States, and the consumption of alcohol has increased over the years (Jones, Chryssanthakis & Groom, 2014). This is due to many different reasons, including social acceptance and negative peer pressure placed on individuals by the media and peers (Teunissen et al., 2014). Alcohol is often viewed as a coping mechanism for social anxiety (Huot, Thrivikaman, Meaney & Plotsky, 2001). Alcohol is sometimes used heavily while in the social setting because it relaxes people, masking the fear of social interaction that many individuals feel. Levels of consumption also increase based on social acceptance (Huot, Thrivikaman, Meaney & Plotsky, 2001). Presenting an intoxicated familiar rat to a sober one increased alcohol intake over time (Maldonado, Finkbeiner, & Kirstein, 2008). This may be due to social acceptance and/or social anxiety (Tomie et al., 2005). The current study examined change in EtOH consumption over time after no social interaction or social interaction with a novel or familiar partner. A two bottle choice limited access paradigm will be used for voluntary consumption of 12% EtOH. Social interaction involved 20 minute exposure to novel or familiar partner in a novel environment. It is hypothesized that after exposure to social interaction with familiar partners, rats will consume more EtOH compared to rats with no social interaction and social interaction with novel partners. This research is important in understanding the relationship between social acceptance and EtOH consumption. Support was received from the Drinko Center at Westminster College. 104 Email: turnhc22@wclive.westminster.edu 153.2 Anthony Sloan Deanna Buffalari Concomitant effects of methylphenidate and ethanol on working and reference memory in a rat model of ADHD A.R. SLOAN, D. BUFFALARI Westminster College There has been an increase in the number of college students who have reported taking Ritalin while they are drinking alcohol. Ritalin and alcohol are being abused by people who are diagnosed with ADHD and have a prescription for Ritalin but are also being abused by people who do not have a prescription. However, not much work has examined how these drugs interact to affect working and reference memory. The purpose of this research was to examine the concomitant effects of methylphenidate (MPH) and ethanol (EtOH) on working and reference memory in a rat model of ADHD. Twelve rats were trained on the Radial Arm Maze task and tested after injections of saline, MPH alone, EtOH alone, and MPH + EtOH. These tests were given before and after a 6-­‐hydroxydopamine lesion to the medial pre-­‐frontal cortex, which was used to model ADHD. Prior to surgery, the combination of MPH and EtOH decreased working and reference memory compared to other conditions, with more moderate effects of EtOH alone. MPH did not significantly affect working and reference memory. Additionally, it is predicted that the number of errors following surgery will be higher across drug treatments. By understanding how the combination of Ritalin and alcohol affects memory, we can better educate youth on the dangers of taking both substances simultaneously. Research Support: Westminster College Drinko Center, Westminster College Psychology Department Email: sloaar22@wclive.westminster.edu 153.3 Jacob Pletz Deanne Buffalari The effects of methylphenidate and ethanol on impulsivity in a rat model of ADHD J.D. PLETZ, D. BUFFALARI Westminster College The percentage of adults co-­‐abusing ethanol and methylphenidate has increased in recent years as adults have more access to methylphenidate through others with a prescription (Darredeau, Barrett, Jardin, & Pihl, 2007). The coabuse of methylphenidate and ethanol can lead to impulsive decisions due to the increases of catecholamines and the cognitive effects of ethanol. Therefore purpose of this study is to examine the effects of methylphenidate (MPH) and ethanol on impulsivity in a rat model of ADHD. Impulsivity will be assessed using a T-­‐maze, which will ask rats to choose between a large delayed reinforcement (correct choice) and a small but immediate reinforcement (incorrect choice). Animals will be assessed in four conditions: MPH, ethanol, their combination, and saline. These conditions will be tested before and after a 6-­‐hydroxydopamine lesion to the mPFC. This study will enhance our understanding of the interactions of MPH and ethanol on impulsivity in those with and without ADHD. Preliminary data suggest that healthy rats injected with ethanol are more impulsive than those injected with the combination, methylphenidate and saline. Drinko Center at Westminster, Westminster College Psychology Department. Email: pletjd22@wclive.westminster.edu 105 154 Sahba Seddighi Matthew Cooper A neurocognitive study of second language learning: learning conditions, memory profiles and outcomes S. SEDDIGHI, A. BRITO, M. SARRETT, H. BOWDEN University of Tennessee, Knoxville Though it is often assumed that immersive experiences are more effective than classroom learning in the acquisition of a new language, the underlying neurocognitive basis of implicit and explicit learning remains an important topic of investigation. In this project, we examine the effects of different pedagogical methods on brain activity in the acquisition of a second language (in this case, Latin). While utilizing a computer-­‐administered program to train and test participants in aspects of Latin, we compare their behavioral and neurocognitive variations in linguistic processing. Participants are divided into two groups: those who are taught using an implicit mode of instruction (mimicking immersion learning) and those taught using an explicit mode (mimicking classroom learning). The resulting linguistic consolidation is assessed via a battery of linguistic tasks and through the use of electroencephalography (EEG) to measure event-­‐related potentials (ERPs) elicited by the learners’ brains when reading sentences that are either correct or that contain a grammatical (syntactic) or lexical (semantic) violation. In addition, we are interested in exploring whether how well a second language is learned in a given environment correlates with participants’ relative strengths of different types of memory. To examine this question, we test declarative and procedural memory strength prior to language training. Thus, we can examine not only which method is more effective overall, but also whether they lead to different neurocognitive outcomes as reflected by ERPs. In addition, we can assess the effect of learners’ memory profiles on both their learning in those conditions and their neurocognition of the newly acquired language. We predict that (1) participants will show different ERP signatures while reading incorrect syntactic and semantic sentences in Latin, that (2) the method of instruction will affect language learning, and finally, that (3) a difference in neurocognitive processing and/or Latin proficiency will be seen with participants’ varying strengths of declarative and procedural memories. Email: sseddigh@vols.utk.edu 155 Alex Schultz Meg Waraczynski Blocking CaV1.3 channels in the sublenticular extended amygdala has a delayed effect on MFB self-­‐stimulation A.V. SCHULTZ, M. WARACZYNSKI University of Wisconsin -­‐ Whitewater Past research done in our lab has shown that the sublenticular central extended amygdala (SLEAc) has a role in the reward efficacy of medial forebrain bundle (MFB) stimulation. Previous work has shown that stimulating D2 dopamine receptors in the SLEAc impairs MFB self-­‐stimulation more than D1 receptor blockade does. Therefore, we are currently exploring the role of D2-­‐mediated cellular mechanisms in MFB self-­‐stimulation. D2 stimulation indirectly blocks calcium currents through CaV1.3 (L-­‐type) somatodendritic calcium channels, which in turn challenges neural excitability. If D2 stimulation acts on MFB stimulation reward via this mechanism, then directly blocking these channels should also impair MFB stimulation reward efficacy. Male Long Evans rats received unilateral stimulation electrodes aimed at the MFB and bilateral guide cannulae aimed at the SLEAc. CaV1.3 channels were blocked ipsilateral and contralateral to the stimulation site with 2.5µg and 5µg of the phenylalkylamine verapamil and 5µg and 10µg of the benzothiazepine diltiazem. Drug-­‐induced changes in the stimulation’s reward efficacy were measured with the rate-­‐frequency curve shift technique. There was no significant effect of drug condition on the day of the drug injection. However, there was a significant and substantial increase in the frequency required to maintain half-­‐maximal responding for the stimulation (i.e., a decrease in the stimulation’s reward efficacy) on the day after injection of 10 μg of diltiazem ipsilateral to the stimulation 106 site. No other drug condition produced this delayed effect. The frequency required to maintain half-­‐
maximal responding returned to baseline levels by the next testing session. The time course of this effect suggests that blocking Cav1.3 calcium currents in the SLEAc ipsilateral to the stimulation site may trigger a delayed but transient change in some cell signaling mechanism that is important to the stimulation’s rewarding effects. The fact that the effect occurred only with ipsilateral injections -­‐-­‐ whereas we have observed substantial effects of other drug injections contralateral to the stimulation site – supports a hypothesis that backpropagating action potentials produced by the MFB stimulation may be important to its rewarding effects. This work was supported by grant #IOS 1050256 from the National Science Foundation to M. Waraczynski and by grants from the UW-­‐Whitewater Undergraduate Research Program and the Biology Research Fund to A. V. Schultz. Email: schultzav03@uww.edu 156 Sneha Gupta Catherine Marie Davis Behavioral Effects of Dopaminergic Drugs in Irradiated Rats S.R. GUPTA, C.M. DAVIS, R.D. HIENZ Washington and Jefferson College Previous studies have shown that head-­‐only proton radiation exposure to rats results in performance deficits in the rat Psychomotor Vigilance Test (rPVT). Two groups of rats emerged based on rPVT performances following exposure: “radiation sensitive” rats and “radiation insensitive” rats. More specifically, sensitive rats showed changes in accuracy and premature responding on the rPVT, while insensitive rats performed like sham-­‐irradiated control rats. Since the rPVT generally tests sustained attention, integrity of the dopaminergic system in these rats was subsequently tested using methods that elicit dopaminergic-­‐mediated behaviors. Dopaminergic receptor agonist-­‐induced yawning occurs following activation of the dopamine D3 receptor; activation of the dopamine D2 receptor inhibits this yawning response. Thus, a dopamine D2/D3 receptor agonist, Quinpirole (0.01-­‐1.0 mg/kg) was used to assess D3-­‐induced yawning and its inhibition by concurrent activation of D2 receptors. A dopamine D2 receptor antagonist, L-­‐741,626, was used to assess changes in D2 receptor levels in radiation sensitive or insensitive rats. The two drugs were used in combination to produce dose response curves, which were compared within groups of sensitive and insensitive rats. Email: guptasr@jay.washjeff.edu 157 Quentin Richardson Chris Goode Method for analyzing c-­‐Fos colocalization in vasopressin and oxytocin cells within the PVN Q. RICHARDSON, M. PAUL, G. DE VRIES Georgia State University Social play behavior in juvenile animals has been shown to have a significant impact on the development of adult social skills. By studying social play behavior in juveniles, we can get a better understanding of social skill development, and a better understanding of social development disorders such as autism spectrum disorder and attention deficit hyperactive disorder. However, little is known about what neural systems regulate social play behavior in juveniles. In this study, we investigate the periventricular nucleus of the hypothalamus (PVN) of juvenile rats to see if it plays a role in social play behavior in juveniles. In this experiment, we devised a method using fluorescent confocal microscopy to analyze the PVN of juvenile rats. Prior to this experiment, juvenile rats had social play interaction, then their brain tissue was stained for vasopressin, oxytocin, and c-­‐Fos, which is an indication for neural activation. With the use of practice tissue, a standard exposure was determined for each fluorescent staining. Furthermore, a method 107 for analyzing and counting vasopressin and oxytocin cells, as well as confirming if these cells are colocalized with c-­‐Fos has been determined. Based on our standard exposure for each staining, the oxytocin and vasopressin cells, as well as c-­‐Fos colocalization can be distinguished through tissue. Currently, micrographs of experimental tissue are being analyzed. In conclusion, we developed a procedure to be ideal for unbiased analysis of the experimental tissue using fluorescent confocal microscopy. Analysis of experimental tissue is not complete, but possible outcomes are the PVN plays a role in social play behavior in juvenile rats, or the PVN does not play a role in social play behavior in juvenile rats. If the PVN does play a role in social play behavior, future studies would be to manipulate the PVN. 158 Damaris Pop Shane Perrine Changes in brain norepinephrine and serotonin levels in an animal model of posttraumatic stress disorder D.E. POP, R.J. KOHLER, M.J. LISIESKI, M.G. BAUER, A.L. EAGLE, S.A. PERRINE Wayne State University Posttraumatic Stress Disorder (PTSD) is a debilitating condition that is characterized by re-­‐experiencing the traumatic event, avoidant behavior, hyper-­‐arousal, and negative cognition and mood. Although extensive pre-­‐clinically and clinically research has been done exploring PTSD, its neurobiology remains to be fully understood. The aim of this study was to examine the effects of single prolonged stress (SPS), an animal model of PTSD, on monoamine levels in the brain. Male Sprague-­‐Dawley rats were exposed to SPS treatment, consisting of a consecutive series of stressors (2 h restraint, 20 min group forced swim, and ether exposure until unconsciousness) followed by a 7 d incubation period. Both SPS and control rats were decapitated after the incubation period and their brains dissected for analysis of monoamines (norepinephrine, NE; serotonin, 5-­‐HT) using high pressure liquid chromatography (HPLC). Regions of interest involved in PTSD were examined: dorsal hippocampus (dHC), intermediate-­‐ventral hippocampus (i-­‐vHC), and amygdala. Results showed significant increases in 5-­‐HT within the dHC and i-­‐vHC and NE within the i-­‐vHC. There were no SPS-­‐induced differences in the amygdala. These data indicate that SPS alters NE and 5-­‐HT levels in key brain regions involved in regulating emotional states in response to environmental stimuli. Email: damarisepop@gmail.com 159 Eileen Cho Virginia Quinan Color categorization in macaque monkeys. E. CHO, E.LEE, B. CONWAY Wellesley College Color categorization differs between languages. However, common color classifications across many languages suggests the existence of innate color categories. To investigate the nature of categorization without the effect of language, we will implement two different paradigms to see whether macaque monkeys, a model for human color vision, have innate color categories. Furthermore, we will investigate how monkey categories compare to human categories. Our first paradigm is modeled after studies in infants. Subjects will detect a colored target stimulus on a differently colored background. The varying colors of the target and the background will be equally sampled between two colors in an isoluminant hue circle in DKL color space (Derrington et al. 1984). The target detection time will be measured as an indicator of categorization. Subjects will react faster to the target if two colors are of different categories than of same categories. Thus, a significant drop in reaction time will illustrate the categorical boundaries between the two colors. Our second paradigm is modeled after adult human studies. Subjects will be 108 shown sets of four colors, also sampled in DKL space in the same manner as the first experiment. In each set, subjects will pick out the color that appears different from the rest. Based on response and reaction times, we will derive the color category boundaries. Since perception of colors depends not only on hue but also on luminance and saturation, further experimentation is needed to explore those effects on the categorization of color in both humans and monkeys. Acknowledgements: Wellesley College Harvard Medical School Claudine Malone '63 Summer Science Research Scholars Gift Sherman Fairchild Foundation Email: echo4@wellesley.edu 160 Melissa Gorham Janet Finlay Effects of NMDA receptor dysfunction in ventral medial prefrontal cortex on a delayed spatial reference memory radial maze task in adult mice M. M. GORHAM, T. V. NGUYEN, R. M. WESTON, A. M. SCHILLER, T. V. NGUYEN, , R. F. KYDD, M. J. MANA, R.W. GREENE, J.M. FINLAY Western Washington University Dysfunction of glutamate N-­‐methyl-­‐D-­‐aspartate (NMDA) receptors may contribute to cognitive deficits in schizophrenia. In the present study, we examined the effects of chronic NMDA receptor dysfunction in the ventral medial prefrontal cortex (VmPFC) on acquisition of a spatial reference memory radial maze (SRM) task as employed by Niewoehner et al. (2007). Localized NR1 gene deletions were induced in the VmPFC of floxed NR1 mice (DEL, n=10) using an AAV-­‐Cre vector; Control mice (CON, n=10) received sham deletions. In the SRM task, food was placed in 3 arms of a 6-­‐arm radial maze at the start of each trial; the location of the food was consistent for each mouse across all trials, but randomly varied between mice. At the start of each trial, each mouse was placed in the central chamber of the maze and allowed to enter any of the 6 arms; once an arm was visited and a mouse returned to the central chamber, the entrance to each of the 6 arms was blocked for 10 sec. At the end of each 10 sec timeout, the arms that had not yet been visited during the trial were made accessible and a mouse could choose to enter one of the remaining unvisited arms. Thus, this task only assessed reference memory errors (RME); that is, entries to arms that were never baited. Mice were tested in 4 trials/each daily session. We found that NMDA receptor dysfunction in the VmPFC had no effect on acquisition of the SRM task. There were no differences in behavior observed in the first session (mean arms visited, DEL = 5.3 arms and CON = 5.2 arms; mean baited arms visited DEL and CON = 3.0; mean unbaited arms visited, DEL = 2.3 arms and CON = 2.2 arms). Mice in both the groups acquired the SRM task over 24 sessions (mean arms visited, DEL = 3.5 arms and CON = 3.6 arms; mean baited arms visited DEL and CON = 3.0; mean unbaited arms visited (that is, RME), DEL = 0.5 errors and CON = 0.6 errors). After the 24-­‐session Acquisition Phase, we rotated the extra-­‐maze spatial cues to confirm the use of such cues in SRM performance by both groups (mean RME during the Rotation session, DEL and CON = 1.8 errors). The lack of significant between-­‐group differences in the SRM task in our study contrasts with recent reports that Grin1DGCA1 mice with NMDA receptor deletions in the dentate and CA1 regions of the hippocampus show impaired acquisition in this task (Bannerman et al., 2013), reflecting different contributions of the VmPFC and hippocampus in the task. Given the well-­‐known contribution of the PFC to working memory, we are currently assessing PFC NR1-­‐deleted mice in a working memory variant of this task. Email: gorhamm@students.wwu.edu 161 Edwin Glueck Jeff Grimm Abstinence and environmental enrichment related changes in Fos expression in rats responding for a sucrose-­‐paired cue E. GLUECK, J. W. GRIMM, J. BARNES, J. KOERBER, D. GINDER, J. HYDE, L. EATON 109 Western Washington University Exposure to environmental enrichment (EE) profoundly reduces sucrose seeking by rats. The present experiment was conducted to examine whether brain Fos levels correlate with levels of cue-­‐reactivity in rats exposed to either acute or chronic EE. The experiment also included rats tested for cue-­‐reactivity after either 1 or 30 days of forced abstinence to examine whether EE and/or Fos effects would vary with the incubation of sucrose craving. METHODS: Fos expression was examined in 18 regions of rat brain following 10 d (2h/d) of sucrose self-­‐administration and a 2h cue-­‐reactivity test on a subsequent day. Prior to this test, rats experienced either 1 or 30 d of forced abstinence and either overnight (acute) or 29 d (chronic) EE. EE consisted of 3 rats housed in a large, multi-­‐level cage with novel toys exchanged 3 times a week. Controls were returned to single housing. Regions sampled across the rostral-­‐caudal extent of the brain included several cortical regions including the hippocampus, divisions of the striatum, and midbrain regions including the VTA. Fos was identified in brain slices using immunohistochemistry. Immunolabeled cells were counted using NIH Image J at 10X magnification within a 250 micrometer square sample of regions identified using an atlas of the rat brain. RESULTS: Fos expression generally paralleled the amount of cue-­‐reactivity. That is, Fos levels were higher in most regions after 30 d of forced abstinence (“incubation of Fos expression”) and were decreased in most regions by either acute or chronic EE. One interesting dissociation was observed between striatal sub regions where acute EE just prior to the first day of forced abstinence reduced Fos expressed in the dorsolateral striatum but not in the core or shell of the nucleus accumbens. CONCLUSION: Further correlational analyses of these data may yield patterns of connectivity predictive of cue reactivity following exposure to control or EE living conditions and/or in either early or late forced abstinence. Supported by NIH DA016285-­‐03 Email: gluecke@students.wwu.edu 162.1 John Georgino Kelly Weixel Utilizing Neurotropic Viruses to Define Circuit Architecture in Rodent Brain J.R. GEORGINO, J.P. CARD Washington and Jefferson College An understanding of neural circuitry is crucial in the analysis of healthy brain function. A neural circuit is a structural unit of interconnected neurons that work together to maintain homeostasis. When comparing neural circuit tracing techniques, transneuronal viral tracing is superior to classic tracing techniques due to the virus’ ability to cross synapses. To characterize the invasiveness of an Alpha herpesvirus,, a rodent model was sterotaxically injected into the well-­‐documented striatal circuit with a novel strain of Herpes Simplex Virus (HSV) that spreads retrogradely through a neural network. . These viruses are DNA viruses whose preferred host are neurons, and cause encephalitis in humans. Rats were then perfused and prepared for immunohistochemistry to identify the infection. Results from this experiment indicated that the substantia nigra pars compacta was severely infected while the globus pallidus external segment was intermediately infected. Infection of the nuclei were then scored as moderate, intermediate, or severe depending on parameters such as dendritic labeling, density, and presence of nuclear invaginations. Understanding the invasiveness of a retrograde tracer such as this one can make it possible to perform dual injection studies concerning an anterograde tracer and a retrograde tracer. Studies were also conducted using this tracing technology to examine the functional organization of the suprachiasmatic nucleus (SCN). HSV was injected into the dorsomedial subfield of the SCN and perfusions and immunohistochemistry was performed following a 48 hr infection period. Analysis of these images revealed the dense commisural projections between the injection site and the contralateral dorsomedial subfield as well as projections between the ventrolateral subfields along with efferent projections to the subparaventricular zone, networks which are poorly characterized. Contralateral paraventricular nuclear labeling was also identified in these studies, indicating the potential presence of efferent networks. However, further studies need to be done to analyze these contralateral projections, which could be revealed by alternative viral tracing tools. Wojaczynski, G.J., Engel, E.A., Steren, K.E., Enquist, L.W., Card, 110 J.P. (2014) The neuroinvasive profiles of H129 (herpes simplex virus type 1) recombinants with putative anterograde-­‐only transneuronal spread properties. Brain Struct Funct. Email: georginojr@jay.washjeff.edu 162.2 Leah Vaughan Kelly Weixel Adolescent high-­‐fructose diet alters metabolism and the HPA axis without exacerbating behavior following adult ischemic stroke L.E. VAUGHAN, C.S. HARRELL, G.N. NEIGH Washington & Jefferson College The recent surge in fructose consumption parallels the rise in obesity, metabolic syndrome, and cardiovascular disease. High-­‐fructose diets (HFD) are associated with insulin resistance, hyperglycemia, and visceral adiposity in animal models, but the effects on neurological disease and behavior remain poorly understood. Preliminary research from this lab has revealed altered metabolism and stress responses, including elevated basal corticosterone, amongst rats fed a HFD throughout puberty. This altered stress response could be associated with neuroinflammatory and cerebrovascular changes that have the potential to affect outcomes after neurological injury. Thus, this study sought to determine the extent to which consumption of a high-­‐fructose diet throughout adolescence compromises adult ischemic stroke outcomes. We hypothesized that a HFD will exacerbate the effects of ischemic stroke to exaggerate altered stress responses and impair neurological and affective behaviors. Adult male rats were fed either standard chow or 55% high-­‐fructose diet from two days post-­‐weaning through adulthood. Rat food consumption, blood glucose, and weight gain were tracked regularly for all animals, fat pad weight was collected from a non-­‐surgical cohort after 10 weeks on the diet. As expected, HFD increased blood glucose, caloric efficiency, and fat pad mass. Middle cerebral artery occlusion (MCAO) or sham surgeries were then performed on two cohorts of rats after 8 (cohort 1) or 10 (cohort 2) weeks on the diet. Neurological testing (open field, sticky dot, rotorod, and grip strength) was performed prior to surgery and again three days post-­‐surgery. While MCAO did worsen performance in the open field, sticky dot, and rotorod, no additional effect of HFD was apparent. Tests of affective-­‐like behavior (open field, social interaction, elevated plus maze, forced swim) were performed two weeks post-­‐surgery. HFD significantly reduced grooming behavior, while both MCAO and HFD significantly altered social interaction behavior. However, the effects of MCAO and HFD did not appear to be additive to exacerbate effects in the social interaction test or other tests of affective-­‐like behavior. Analysis of corticosterone levels at baseline and following a forced swim test (FST) by enzyme-­‐linked immunosorbant assay indicated that while HFD significantly increased baseline corticosterone, there was no interactive effect of HFD and MCAO on corticosterone at baseline or after HFD. Ongoing studies are assessing the impact of HFD on additional outcomes after MCAO, including infarct size, cerebrovascular remodeling, blood-­‐brain-­‐barrier permeability, and neuroinflammation. Email: vaughanle@jay.washjeff.edu 163 Chad Wagner Kevin Crisp Development of wireless, passive electrophysiological technologies C.L WAGNER, A.W. BIGELOW, C.J. STEWART, K.M. CRISP St. Olaf College Modern techniques for electrophysiological stimulation require that an organism either be tethered to a long wire (Anderson et al., 2006) or carry a battery (Zhang et al., 2011) to power the stimulator. Wires limit mobility, while batteries can be large, generate excess heat, and occasionally need to be replaced. These techniques also risk injury and infection, making them suboptimal to study natural animal 111 behaviors. We are trying to create a wireless, battery-­‐free, electrophysiological stimulator that could be implanted in an animal to provide a controllable stimulation to a signal neuron or ganglia. We developed a resonant circuit that could receive a signal from a near-­‐field, radio frequency source and deliver a measurable stimulus to an L.terrestris (common earth worm). While effective, our circuit must still be miniaturized, and a higher efficiency tuned antenna must be developed. We must also create our stimulator in a manner that does not interfere with tissue or cause an immune response. One way we seek to accomplish this is with development of self-­‐sealing, bio-­‐invisible cuff electrodes (Korivi et al., 2011). Our cuff electrodes were not only able to record neural impulses non-­‐invasively and with high fidelity, but were also able to aid in the deliverance of an isolated stimulus. If we can miniaturize our antenna, stimulator circuit, and cuff electrodes, it would allow us to wirelessly stimulate select neurons while affording the organism freedom of motion. This accomplishment would provide greater accuracy (Arfin et al., 2009) when performing studies on animal behavior and could serve as a first step towards the development of an advanced era of microscopic medical devices Email: wagnerc@stolaf.edu 164 Evan Meiman Lora A. Becker Behavioral effects of magnesium oxide versus magnesium L-­‐threonate diet in adolescent sprague dawley rats E.J. MEIMAN, M.A. ERMLER, L.A. BECKER, R. WAWORUNTU, B. BERG University of Evansville Magnesium is an essential mineral that serves as an intracellular cofactor in enzymatic reactions and has an important regulatory role of gating of the N-­‐methyl-­‐D=aspartate receptor which is involved in learning and memory processing). The purpose of this study was to assess the impact of Magnesium Oxide (MgO) versus Magnesium L-­‐Threonate (MgT) during normal early postnatal development on anxiety-­‐related behaviors in rodents. Newly weaned Sprague-­‐Dawley rats, postnatal day (PD)21, were fed a diet that either contained 0.1% MgO or 0.1% MgT. No significant differences were found for body weights or food intake across the study. Open field (OF) exploration of PD28 or PD35 rats resulted in a Diet X Gender interaction (F= 9.46, p < 0.05) wherein female rats on MgO spent more time moving around the OF but female rats fed MgT displayed more rearing frequency (F=4.11, p<0.05) and duration (F=3.83, p<0.05). There was a interaction of Diet X Age X Area within the OF Arena (F=2.66, p0.05) wherein rats fed MgT spent more time sitting away from the walls than rats fed MgO, which increased from PD28 to PD35. Preliminary analysis of behavior in an acoustic startle (AS) chamber suggests that rats fed MgT tended to have a faster habituation curve to the startle stimulus and lower velocity of startle than rats fed MgO. Overall, MgT fed rats performed more investigative-­‐like behavior and less anxiety-­‐like behavior in the OF arena than rats fed MgO. Mead Johnson Pediatric Nutrition Institute grant to Lora Becker Email: em162@evansville.edu 165 Galen Chuang Bevil Conway Processing and displaying fMRI data by color selectivity. G. CHUANG, B.R. CONWAY Wellesley College Functional magnetic resonance imaging, or fMRI, measures brain activity over time by detecting increased blood flow to certain areas. Processing involves motion correction, registration of functional data to anatomical MRI scans, smoothing, creating contrasts, and upsampling. The functional data can then be mapped computationally onto high resolution, three-­‐dimensional surfaces. After processing, fMRI data is typically displayed based on significance values from statistical t-­‐tests; these show the probability that the 112 same results could be obtained again. However, our project sought to display fMRI data with selectivity indices, or which stimulus each voxel responds to the most strongly. By manipulating the fMRI processing stream, we developed techniques to calculate selectivity indices and display them on a cortical surface. Our final goal is to display the selectivity index of each voxel using the color that voxel was most selective toward, allowing spatial patterns and groupings of colors to be more easily detected. Once implemented, this may give clues to understanding the hierarchal organization and evolution of the visual cortex. Email: gchuang@wellesley.edu 166 Dana Cobb Leslie Sargent Jones The value of the IMPULSE experience for non-­‐neuroscience majors. D. COBB, E. ARTZ, H. JOHNSON, B. MANSKY, A. ROSSI, D. RUSSELL, C. ESTER, T. SASSER, R. SLEDGE, W. STEVENS Appalachian State University The journal IMPULSE was created over a decade ago to provide undergraduates with an opportunity for an authentic experience reviewing original neuroscience submissions, as well as to serve as an outlet for publishing undergraduate research. While initially designed with the neuroscience major in mind, many of the students working with the journal are not in neuroscience or related majors. For example, of the 21 reviewers at one of the Reviewer Training Sites, four are targeting neuroscience careers, seven are pre-­‐
medical students, while the other majors are math (3), geology (2), journalism (1), religious studies (1), and undecided (3). They all joined to learn how to review and write science papers; they were indifferent to the discipline of the vehicle used for that training. In support of this are results from a survey conducted in 2012-­‐13 with previous journal participants, indicating the value of the experience to non-­‐neuroscience majors. As reported on the data from that survey, the majority of respondents felt it was useful for their professional writing and literature research skills, and that this applied to their long-­‐term goals regardless of their field. While the outreach to recruit participants has historically highlighted the relevance of the journal experience for students pursuing careers in neuroscience, the past decade has shown that other majors are benefiting from this opportunity and that it should be extended intentionally to students in other fields. 167 Leanne Harris Page Baluch Engaging students in brain awareness through hands on science L. HARRIS, A. GONZALES, J.J. FAUST, I. SINAKEVITCH, D.P. BALUCH Arizona State University Engaging a student’s interest in STEM education requires not only great mentorship but activities that can capture their attention and stimulates their interest to learn more. Hands on Science is an annual event held at Arizona State University which encourages young scientists to pursue a career in STEM fields such as neuroscience by giving them the opportunity to experience cutting edge research in a university environment. Students, especially women, minorities and those with disabilities, are invited to attend this one day event to tour various labs at ASU and participate in hands on activities ranging from investigating crime scenes using DNA fingerprinting, engaging in alternative energy experiments using anaerobic chambers, observing insect behavior studies, viewing meteorites and images taken by the Lunar Reconnaissance Orbiter camera to mounting slides and scanning images of neurons with laser based microscopes. The W.M. Keck Bioimaging lab, within the School of Life Sciences, specifically focuses on the Brain Awareness portion of the event by providing multiple stations that give students the opportunity to learn how image based neurobiology materials can be made accessible to the visually impaired, how behavioral neuroscientists trace neurons to observe changes in plasticity in response to 113 stress, and how fluorescently labeled brain sections and neural cultures are imaged using instrumentation such as the laser scanning confocal microscope. Students share their microscope images online at the ASU Hands on Science Facebook page (www.facebook.com/ASUhandsonscience) which provides a portal for ongoing education and gives them the opportunity to share their experience with friends and family. https://www.awis-­‐caz.org/hos.html Email: lmwillis@asu.edu 168 Hillary Blakeley Hillary Blakeley Effects of a gamified classroom on student self-­‐efficacy. H.J. BLAKELEY, B.J. BLAKELEY Merrimack College Gamification refers to the use of game mechanics in a non-­‐game settings. Though commonly misunderstood as using games in the classroom, gamification has been used extensively in business and marketing as well as in education. It often contains components like achievements, badges, “leveling up,” reward points, and specific tasks or challenges to be met. This poster reflects the author’s experience teaching a “gamified” Honors Introduction to Psychology class, designed as a series of levels that students progressed through as the semester proceeded. Each level had to be passed by successfully completing a task, and tasks differed throughout the semester generally becoming more demanding of time, effort, and skill. Students were explicitly told that they might have to attempt a task multiple times, but that there was no penalty regardless of how many failures preceded the successful attempt. Final grades in the course were determined entirely by what level the student attained by the end of the semester. Because gamification has been associated with higher student self-­‐regulation, the authors used the MSLQ as a pre-­‐ and post-­‐test measure for motivation. By the end of the semester, students’ scores on self-­‐efficacy for learning and performance, one of the sub-­‐scales of the MSLQ, had increased significantly from pre-­‐test levels (p<.01). This suggests that classroom gamification may empower students to take control of their own success and progress through the course. This work was supported by a Merrimack College Faculty Development Grant. Email: blakeleyh@merrimack.edu 169 Nicholas Hardy Ellen M. Carpenter UCLA neuroscience outreach to LA schools N.F. HARDY, D. ALEXANDER, M. DESALVO, R. ROMERO-­‐CALDERON, C.A. GHIANI, W. GE, C.E. EVANS, J.B. WATSON, E.M. CARPENTER University of California Los Angeles UCLA's Interdepartmental Program in Neuroscience and the Brain Research Institute conduct major outreach activities throughout the year through Project Brainstorm and Brain Awareness Week. Both of these activities aim to promote an interest in neuroscience and higher education to underserved schools around the greater Los Angeles Area. Project Brainstorm (PB) is an outreach course offered to 3rd and 4th year undergraduate majors in neuroscience at UCLA. The course offers undergraduates an opportunity to design and implement an interactive classroom lesson on neuroscience with the objective of providing K-­‐12 students a framework in neuroscience including brain structure, features of a neuron, and basic principles of synaptic communication. In the first part of the course, students select a topic and develop their presentations while receiving feedback from a group of faculty, staff, graduate students, and their peers. This year's topics included reflexes, the visual system, pain perception, motor movement, and olfaction. The undergraduates are then invited into classrooms primarily in Title 1 K-­‐12 schools in the Los 114 Angeles (LA) area to present their lesson plans and engage younger students in a variety of interactive neuroscience activities. UCLA's Brain Awareness Week (BAW) is an annual event that brings students from LA schools to campus for a day of neuroscience related activities. The event is organized by graduate students from the Neuroscience Interdepartmental Ph.D. program and is staffed with over 60 volunteers from the graduate and undergraduate neuroscience programs, the David Geffen School of Medicine, and UCLA faculty. Over the course of the week, we hosted 300 students between 4th and 12th grade from five LA schools. Each day began with the visiting students rotating through the following six interactive stations: 1) Brain anatomy, 2) Homunculus mapping, 3) Vision-­‐altering goggles, 4) Human brain specimens, 5) Animal brain specimens and evolution, and 6) Brain injury. Students then participated in lab tours that included hands-­‐on demonstrations such as extracting DNA from cheek cells and viewing aplysia or drosophila and in campus tours. To evaluate the efficacy of BAW and PB in increasing neuroscience knowledge and inspiring an interest in science in the students we administered pre-­‐ and post-­‐evaluation forms for the students to complete. Preliminary data analysis indicates that students, across all age groups, knew significantly more basic neuroscience after their BAW and PB experiences than before. Further, many students indicated a more positive attitude toward science after their visit. Email: nhardy@mednet.ucla.edu 170 Rylie Hightower Elba Serano MICAL2: A necessary ectoderm development modulator R. HIGHTOWER, E. BATES New Mexico State Univ. During embryonic development, ectoderm differentiates into external tissues and structures of a developed human, such as skin, hair, nails, and teeth. The CNS also develops from differentiating ectoderm. Currently, ectodermal dysplasia (ED) is considered a hereditary, congenital disorder that results in abnormalities of those external tissues and structures. The analysis of one specific family shows that some members have ED and some members have autism. MICAL2 is a gene that has shown to be mutated in each family member that has ED or autism. Previous work shows MICAL2 is needed for axon guidance and actin depolymerization. This study was to determine the role MICAL2 plays in ED and Autism. Site-­‐directed mutagenesis is being used to clone and replicate the affected family members’ mutation. Further work will use “Knockdown” and “Rescue” methods to “knockdown” MICAL2 function in wildtype cells where cell function can then be assessed. We predict that normal cell developmental function will occur with a wildtype MICAL 2 rescue but normal cell development will cease with a mutant MICAL2 rescue. The exact abnormalities that will occur on the cellular level with the mutant MICAL2 rescue are unknown and will be assessed at the time of observation. Email: swimmer1@nmsu.edu 171 Kathryn Sanchez Elba Serano Amyloid beta truncated fragments: Relevance for Alzheimer's disease pathogenesis K. SANCHEZ, J. GHISO New Mexico State Univ. Alzheimer’s disease (AD) is the most common form of dementia, which currently affects 5 million Americans. Amyloid beta (Abeta) is the main component of parenchymal plaques and vascular deposits, two of the major neuropathological lesions of AD. In addition to full-­‐length Abeta species, biochemical and proteomic analysis of AD deposits reveal high degree of Abeta heterogeneity at both N-­‐and C-­‐terminal ends likely resulting from the local action of multiple proteolytic enzymes. Interestingly, many of these fragments -­‐particularly those truncated at the C-­‐terminus-­‐ are also normal components of cerebrospinal fluid and have biophysical properties supporting their association with brain clearance mechanisms. 115 Conversely, very little is known about the role of N-­‐terminally truncated fragments, which we postulate are contributors to the disease pathogenesis. To better understand their properties and evaluate their potential association with the brain lesions, we generated antibodies specifically recognizing N-­‐terminal epitopes and performed biochemical and immunohistochemical analyses. The tendency of N-­‐terminal truncated species to rapidly aggregate and fibrillize together with their topographic localization at the core of amyloid plaques in mouse transgenic models support the notion of their participation in the mechanism of amyloidogenesis. Email: ktsanch@nmsu.edu 172 Tiffany Faull Mary Morrison Purkinje neuron developmental markers in vivo and in vitro. T.N. FAULL, L.M. ROBERTSON, A.R. DEMCHAK, M.E. MORRISON Lycoming College The cerebellum coordinates movement and balance. It is composed of only a few cell types including Purkinje neurons, whose dendrites receive inputs via synapses from granule cell parallel fibers and olivary climbing fibers. The Purkinje neurons integrate these inputs, calculate motor error, and send corrective signals through their axons which synapse onto the deep cerebellar nuclei. The early development of Purkinje neuron axons and dendrites is the subject of this study. The growth of Purkinje neurons in the mouse can be divided into several stereotyped stages. In the prenatal stage, the Purkinje cells are arranged in masses within the cerebellar anlage. From postnatal day 0 to postnatal day 3 in vivo, the cells extend numerous, very simple processes. As the cell continues to develop during the first two postnatal weeks, these processes recede back into the cell, and apical dendrites begin to appear. By the second or third postnatal week, the Purkinje neurons have a highly branched dendrite studded with spines. This series of developmental changes is recapitulated in cell cultures made from neonatal mouse cerebella, with a slight time delay of a few days as the cells recover from the cell dissociation process. Knowing more about the nature of the early Purkinje cell processes could help in the design of treatments to support Purkinje cell regeneration after injury or in the face of cerebellar ataxias. Our goal is to characterize the primitive processes of Purkinje cells in early postnatal mice and in cerebellar cultures derived from these mice: are they axonal, dendritic, both, or neither? In this study, immunohistochemistry of cryostat sections and immunocytochemistry of cultured cerebellar cells was used to establish the locations of several proteins in developing Purkinje neurons, including calbindin D28k, MAP2 (a dendritic marker), and neurofilament H (an axonal marker). Email: fautiff@lycoming.edu 173 Kofi Boateng James Dearworth The Red-­‐eared Slider Turtle: A model for neurodegenerative disease and conservation biology K. BOATENG, R. LAROSA, J. DEARWORTH, M. ROTHENBERGER Lafayette College The red-­‐eared slider turtle (Trachemys scripta elegans), originating from the Mississippi River Basin, is an invasive species present in the Lehigh Valley of Pennsylvania and around the world. Possible reason for its success is possession of a tolerant nervous system that can function under environmental extremes such as anoxia. This study examined turtle populations and the environment, while measuring environmental parameters, to determine if the amount of oxygen in water could be responsible for making the red-­‐eared slider so successful compared to other species. Water sampling and trapping were conducted at sixteen locations from May 2013 to August 2014 within various parts of Pennsylvania 116 (including a pristine conservation site), Maryland, and Michigan. Environmental measures collected included oxygen levels, pH, air and water temperature, salinity, total dissolved solutes, nitrate, phosphorous, turbidity, and conductivity. Turtles were captured by hooped nets and/or photographed at sites to confirm their identities. Data were analyzed using non-­‐metric multidimensional scaling (MDS) ordinations using PC ORD version 5.0 software. The distance between samples in MDS plots reflected relative similarities in environmental and turtle species compositions; r2 cutoff value equal to 0.2 was used to detect vectors of significance for environmental gradients; and r2 cutoff value equal to 0.1 was used to detect significance for turtle species compositions. Analysis of the data showed that red-­‐eared sliders were most frequent when oxygen concentrations in water were low. Turtles also were most visible when air temperature was higher than temperature of water. Of the sites surveyed in Pennsylvania, few turtles were seen or caught at locations having high oxygen concentrations and low saline levels (e.g., Bushkill Creek, Pennsylvania, 40.695251˚, -­‐75.211126˚). Data suggest that environmental parameters do indeed impact species distribution and the ability of red-­‐eared sliders ability to thrive. Sites with greater abundance of red-­‐eared sliders have lower oxygen levels, higher conductivity, and more solutes dissolved, which may explain tolerance of physiology and environmental adaptability of red-­‐eared sliders. The findings support use of the nervous system of the red-­‐eared slider as model for study on how to improve the tolerance levels of human nervous tissue to anoxic conditions that might occur during neurodegenerative diseases (e.g., stroke). Email: boatengk@lafayette.edu 117 Althoff, Alyssa. 67.2 Amato, Michele. 128 Anderson, Maxwell. 141 Andrews, Leigh. 127 Annand, Kathryn. 100 Austin, Paul. 53 Ayabe, Asia. 46.1 Barlow, Molly. 20 Barnard, Elyse. 68 Barnes, Hensley. 124 Barrientos, Alicia. 74.2 Behling, Stuart. 98 Bisnauth, Subrina. 79 Blakeley, Hillary. 168 Boateng, Kofi. 173 Brockway, Emma. 140.1 Buck, Jordan. 143 Buirkle, Julia. 108 Burnham, Veronica. 21 Campbell, Brett. 147 Candal, Raquel. 131 Childs, Ariel. 102.3 Cho, Eileen. 159 Christensen, Jennifer. 115 Chuang, Galen. 165 Claudel, Sophie. 59 Cobb, Dana. 119 Cobb, Dana. 166 Comfort, Nicole. 132 Cone, Katherine. 104 Connell, John. 142 Cottrell-­‐Cumber, Sarah. 11 Crawford, Samuel. 66.1 Cruz, Jayson. 114 Curran, Maura. 107 Curtiss, Sally. 85 Dairaghi, Leigh. 109.1 Day, Marilyn. 41 DeSouza, Andre. 103 Dixon, Paul. 62.1 Dominah, Gifty. 72 Dominguez, Michelle. 64 Doring, Monica. 137 Duncan, Spencer. 33 Dunkerson, Jacob. 90 Ealey, Ashley. 25.1 Eck, Samantha. 129.2 First Author Index (Last, First. Board) Engel, Shaydel. 150 Faull, Tiffany. 172 Feinstein, Max. 135 Ferro, Austin. 80.2 Fettinger, Natalie. 82 Fish, Kyle. 31 Foster, Benjamin. 34.1 Fraser, Kurt. 17 Freedman, Jason. 138 Gehring, Bradley. 40.1 Gentchev, Monica. 91 Georgino, John. 162.1 Gerlach, Matthew. 121 Ghoweri, Adam. 69 Gifford, Janace. 120 Glassman, Caleb. 44 Glueck, Edwin. 161 Goldstein, Jenna. 145 Gorham, Melissa. 160 Grisham, Alanah. 25.2 Gupta, Sneha. 156 Hamilton, Kelly. 122.2 Hamitlon, Sarah. 58.2 Hanten, Brandon. 144 Hardy, Nicholas. 169 Harmon, Katherine. 12 Harris, Brianna. 112.2 Harris, Leanne. 167 Hathaway, Kathryn. 146 Hazim, Manuel. 54.2 Herdegen, Samantha. 152 Hewes, Kelly. 81 Hightower, Rylie. 170 Hossain, Mir Shanaz. 29 Huang, Johnny. 19 Huffman, Heather. 36 Huffman, Lucas. 10 Hughes, Elizabeth. 30 Huynh, Kimberly. 101 Hyde, Alec. 70 Jackson, Kelsey. 50.2 Jang, Eric. 22 John, Caitlin. 136 Juras, J. Anna. 60 Kalidindi, Anisha. 112.1 Kay, Yuni. 46.2 Kechner, Megan. 8 118 Kelly, Kaela. 118.2 Kershberg, Lauren. 66.2 Kim, Juyun. 61 Kish, Eszter. 106 Knabe, Melina. 86 Laufmann, Rachel. 75 Laurenzo, Will. 62.2 Lefevre, Anna. 105.2 Lemma, Eyerusalem. 116 Leonard, Joshua. 28 Lewis, Arille. 43.2 Li, Ranran. 43.1 Liao, David. 111 Lincoln, Cassie. 24 Lopez, Ariel. 15 Lozier, Nicholas. 40.2 Luckett, Kathleen. 49 Luu, Dan. 96.1 Lyons, Ashley. 105.1 Mahajan, Ashley. 37.2 Mathur, Nirav. 67.1 McDonnell, Bobby. 27.2 McDowell, Lana. 129.1 McNamara, Tanner. 110 Meiman, Evan. 164 Melendez, Keyshla. 77.2 Melendez, Zahra. 77.1 Menosky, Megan. 57 Miller, Dylan. 92 Minnick, Kyle. 83 Mitchell, Bridget. 102.1 Moctezuma, Cloe. 140.2 Moncion, Jenny. 74.1 Monuszko, Karen. 58.1 Mootz, John. 130 Mortimer, Emily. 134 Moyer, Andrew. 34.2 Necarsulmer, Julie. 65.2 Newton, Scott. 47 Niro, Biagio. 37.1 Oliver, Khallyl. 71.2 Ordoobadi, Alexander. 84 Pandian, Ashvini. 123 Pearce, Mary. 126.1 Pham-­‐Lake, Camille. 71.1 Phan, Jessica. 14 Phillips, Matthew. 113 Pletz, Jacob. 153.3 Pop, Damaris. 158 Powers, Emily. 5 Pritchard, Amy. 94 Quesada, Pompeyo. 39 Redwine, McKenna. 54.1 Rice, Sharena. 26 Richardson, Quentin. 157 Risi, Cristina. 50.1 Roberge, Kayla. 27.1 Rogers, Josh. 149 Rojas, Jennifer. 52 Rose, Melissa. 139 Ross, Deanna. 99.2 Rotolo, Renee. 9 Rus, Susan. 95 Russo, Craig. 1 Sajjad, Sana. 78 Sakong, Tae-­‐Hyun. 51 Salois, Garrick. 48 Sanchez, Kathryn. 171 Sangaard, Simon. 148 Santos, Maribel. 35 Sautter, Dana. 55 Schlussel, Maura. 23 Schultz, Alex. 155 Schultz, Joseph. 99.1 Schurter, Brandon. 73 Seamon, Kimberly. 102.2 Searles, Madeleine. 89 Seddighi, Sahba. 154 Sewell, Emily. 88 Shoats, Michael. 42 Sloan, Anthony. 153.2 Smallwood, Melissa. 38 Smith, Sarah. 96.2 Sosnowik, Shayna. 97 Srinivasan, Vinay. 65.1 Stafford, Jacob. 3 Stone, Sophia. 118.1 Strand, Helen. 109.2 Stubbeman, Bobbie. 16 Swygart, David. 13 Szolusha, Kerri. 18 119 Tarter, Alexis. 151 Thibault, Mackenzie. 7 Thompson, Cort. 93 Tibbets, Lila. 125 Tishler, John. 122.1 Toker, Nick. 80.1 Turano, Alexandra. 126.2 Turner, Haley. 153.1 Upright, Nick. 6 VanDerhoef, Daniel. 2 Vaughan, Leah. 162.2 Vinzant, Nate. 76 Voigt, Thomas. 45 Wadia, Varun. 87 Wagner, Chad. 163 Wakim, Kathryn-­‐Mary. 56 Wetherell, Joseph. 32 White, Alexandria. 4 White, Cassidy. 133 Willner, Megan. 63 Wise, Bradley. 117 

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