一般演題（ポスター）

一般演題（ポスター） Schizophrenia・Developmental Disorders

2P-47 Translational regulation by the neuronal RNA binding protein Elavl2 in the brain Ohtsuka Takafumi¹，YANO MASATO²，KOYA IKUKO¹，BANNO SATOE¹，SHIBATA SHINSUKE¹，OKANO HIDEYUKI¹ ¹Department of physiology, Keio university school of medicine，²Department of 2nd Anatomy, Niigata university Neuronal Elavls（nElavl）are the first defined neuron specific RNA binding protein（RNABP）, implicated in the nervous system development, making spatialmemory or RNA regulation in drug induced seizure brain. However, their roles have not fully been elucidated, in part due to their in vivo handful targets and redundancy. Here we focused on a specific member of nElavls, Elavl2, which mRNA is expressed in the earliest stage in developing cortical neuronsamong other nElavls. Our histological analysis by Elavl2 specific antibody revealed that Elavl2 protein showed unique expression patterns in the adult mouse brain, especially inhibitory neuron in hippocampus compared with othernElavls. To understand a comprehensive role of Elavl2, we generated genome-wide Elavl2-RNA binding map on the mouse embryonic brain by using HITS-CLIP,High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation methods. We are now validating Elavl2 specific RNA targets with Elavl2 knock-out mouse and this genome-wide Elavl2-RNA binding map, biological functions of Elavl2 by Gene Ontology and in vivo Elavl2 binding motif by CIMS（Crosslinking induced mutation site）analysis, detecting the in vivo Elavl2 binding motif in single nucleotide resolution. In addition, we are also analyzing the mechanisms of Elavl2 function in the brain, especially translational control of target mRNAs with Ribosome profiling analysis. Ribosome profile using Elavl2 KO mice revealed specific RNA targets of Elavl2 in the brain.Lastly, we will discuss how Elavl2-RNA targets link to brain complexity usingthese our two layered in vivo foot printing analysis.		2P-48 Overactivation of the VPAC2 receptor during postnatal brain maturation induces changes in synaptic proteins and selective alterations in prepulse inhibition in mice Ago Yukio^1,2，Hayata Atsuko^1,3，Kawanai Takuya¹，Hashimoto Hitoshi^1,3，Waschek James² ¹Lab. of Mol. Neuropharmacol., Grad. Sch. of Pharm. Sci., Osaka Univ.，²The Semel Inst. and Dept. of Psychiatry, Univ. of California, Los Angeles，³United Grad. Sch. Child Dev., Osaka Univ. Kanazawa Univ. Hamamatsu Univ. Sch. Med. Chiba Univ. ＆ Univ. Fukui Clinical studies have shown that microduplications at 7q36.3, containing VIPR2, confer significant risk for schizophrenia. VIPR2 gene encodes the VPAC2 receptor for VIP（vasoactive intestinal peptide）and PACAP（pituitary adenylate cyclase-activating polypeptide）. Lymphocytes from patients with these mutations exhibited higher VIPR2 gene expression and VIP responsiveness, but mechanisms by which overactive VPAC2 signaling may lead to these psychiatric disorders are unknown. Here we aimed to determine in a C57BL/6 mouse model if the VIPR2-linkage to mental health disorders might be due to overactive VPAC2 receptor signaling during postnatal brain maturation by daily administration of the highly-selective VPAC2 receptor agonist Ro 25-1553 from postnatal day 1（P1）to P14. Western blot analyses on P21 revealed significant reductions of synaptophysin and postsynaptic density protein 95 in the prefrontal cortex, but not in the hippocampus, in Ro 25-1553-treated mice. The same postnatally-restricted treatment resulted in a disruption in prepulse inhibition of the acoustic startle in adult mice. No effects were observed in locomotor activity, sociability in the three-chamber social interaction test, or fear conditioning or extinction. In addition, Ro 25-1553 and VIP, but not PACAP, caused reductions in total numbers and length of neuronal dendrites and length of axon in mouse primary cortical neurons. These results suggest that overactivation of the VPAC2 receptor in the postnatal mouse results in a reduction in synaptic proteins in the prefrontal cortex and selective alterations in prepulse inhibition. These findings imply that the VIPR2-linkage to mental health disorders may be due in part to overactive VPAC2 receptor signaling during a critical time of synaptic maturation.

2P-49 Microglial property changed in a maternal immune activation animal model with relevance to schizophrenia. Yamada Kohei¹，Takahashi Taro²，Mundalil Vasu Mahesh²，Suzuki Katsuaki^1,2，Iwata Yasuhide²，Wakuda Tomoyasu²，Matsuzaki Shinsuke^3,4，Katayama Taiichi⁴，Mori Norio^1,2 ¹Research Center for Child Mental Development, Hamamatsu Univ. School of Medicine.，²Dept. of Psychiatry, Hamamatsu Univ. School of Medicine，³Dept. of Anat. and Neurosci., Osaka Univ. Graduate School of Medicine，⁴Dept. of Child Development and Molecular Brain Science, United Graduate School of Child Development, Osaka Univ. Schizophrenia is a serious psychiatric disorder with a disabled neurodevelopmental basis. Inflammatory and immunological events interfering with brain development are discussed as one of causes of schizophrenia. In our analysis of post-mortem brain, mRNA expression of PBR/TSPO, which is known as microglial activation marker, increased in patients with schizophrenia. Based on this an aberrant neuro-immune system and epidemiological study, maternal immune activation（MIA）as a neurodevelopmental animal model with high validity for schizophrenia has been developed. However microglial property in MIA is unclarified well. The goal of this study is to investigate whether microglia is activated, and then microglial activation is a neurobiological correlate to the altered behavior in the MIA model. In the present study, MIA was induced in pregnant SD rats by injecting intraperitoneally 20 mg/kg poly I：C 2 times per a day at gestational day 13. We examined the number and morphology of microglia in prefrontal cortex（PFC）and hippocampus（HIP）at 4 or 8 weeks old by immunohistochemical stain for Iba1. And we checked mRNA expression of microglial activation markers（IL6, CD68, CD86, IL1b, IL10）in PFC and HIP at 4 or 8 weeks old using real time qPCR. As a result, the cell body of microglia at 8 weeks old in MIA model became bigger and their processes was thicker than control. Although the number of microglia in PFC was not different from control in PFC. On the other hands, the mRNA expression of PBR was upregulated in HIP in MIA at 8 weeks old. And mRNA expression of CD86, which is known as M1 marker, increased in PFC and HIP at 4 and 8 weeks old. These results suggested that microglial property was changed in MIA model and might support the hypothesis that MIA contributes to microglial activation in the offspring. We are examining whether microglial activation relate to abnormal behavior in MIA.		2P-50 Activation of prefrontal dopamine system by attention deficit/hyperactivity disorder drugs improves prenatal valproic acid-induced behavioral abnormalities in mice Hara Yuta¹，Taruta Atsuki¹，Katashiba Keisuke¹，Higuchi Momoko¹，Higashino Kosuke¹，Hashimoto Hitoshi^1,2，Matsuda Toshio³，Ago Yukio¹，Takuma Kazuhiro^2,4 ¹Lab. Mol. Neuropharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ.，²United Grad. Sch. Child Dev., Osaka Univ. Kanazawa Univ. Hamamatsu Univ. Sch. Med. Chiba Univ. ＆ Univ. Fukui，³Lab. Med. Pharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ.，⁴Dept. Pharmacol., Grad. Sch. Dent., Osaka Univ. Rodents prenatally exposed to valproic acid（VPA）are used as animal models of autism spectrum disorder（ASD）. We have also shown that male mice prenatally exposed to VPA at embryonic day 12.5 display ASD-like behavioral abnormalities such as social interaction deficits and recognition memory impairment at 8weeks old. More recently, we have demonstrated that the prenatal VPA exposure causes hypofunction of prefrontal dopamine（DA）system in male mice. The finding implies that hypofunction of prefrontal DA system is associated with behavioral abnormalities observed in the prenatal VPA-exposed mice and activation of prefrontal DA system may result in treatment of the abnormal behaviors. We have found that the attention deficit/hyperactivity disorder（ADHD）drugs methylphenidate（MPH）and atomoxetine（ATX）enhance prefrontal noradrenaline（NA）and DA functions in mice. Thus, the present study examined the effects of ADHD drugs on abnormal behaviors in VPA-treated mice. Chronic, but not acute, treatment with MPH or ATX for two weeks improved social interaction deficits and recognition memory impairment. These drugs also improved the decrements in dendritic spine density in the hippocampus, prefrontal and somatosensory cortices. Furthermore, the improvement of behavioral abnormalities by ATX was blocked by the DA-D₁ receptor antagonist SCH39166 or the DA-D₂ receptor antagonist raclopride, but not by the α₂-adrenergic receptor antagonist idazoxan. These results suggest that ADHD drugs improve VPA-induced abnormal behaviors via activation of DA-D₁ or DA-D₂ receptors. Furthermore, the finding supports that hypofunction of the prefrontal DA system is associated with behavioral abnormalities in VPA mice.

2P-51 Analysis for Mechanism of Autism spectrum disorder via serotonin transporter dysfunction Sato Hiroki¹，Matsuzaki Shinsuke^1,2，Iwata Keiko³，Miyoshi Ko¹，Takamura Hironori¹，Amano Genki¹，Han Sarina¹，Matsuzaki Hideo³，Katayama Taiichi¹ ¹Molecular Brain Science, United Graduate School of Child Development,，²Anatomy and Neuroscience, Graduate School of Medicine,，³Research Center for Child Mental Development, University of Fukui Autism Spectrum Disorder（ASD）is a pervasive development disorder characterized by（1）severe and sustained impairment of social interaction and communication, and（2）restricted or stereotyped patterns of behavior and interest. Since this disorder is thought to be a risk of secondary psychiatric disorders such as depression, the mechanism of ASD is being researched all over the world.Changes in serotonin transporter（SERT）function and expression have been implicated in autism. Our colleagues recently reported decreased SERT levels throughout the brains of autistic individuals compared with controls by using PET. Then we screened the SERT interacting proteins which may affect the function of SERT by IP-MS methods. To narrow down the candidates, we measured their mRNA expression levels in lymphoblast cells of autistic and control individuals by real-time RT PCR and Factor X showed significant increase in the autistic individuals（P＜0.05）. These data suggest the involvement of Factor X in autism via the SERT dysfunction. In this presentation, we show the results of Factor X related to SERT function.		2P-52 Effects of prenatal exposure to a sigma-1 receptor antagonist on behavior and neuronal morphology in rat offspring Yamamoto Toshifumi^1,2，Hirata Akira¹，Yamamoto Hideko² ¹Lab. of Mol. Psychopharmacol., Grad. School of Nanobiosci., Yokohama City Univ.，²Addictive Substance Project, Tokyo Metropolitan Inst. of Medical Sci. Sigma-1 receptors（Sig-1R）have been implicated in the regulation of neuronal differentiation and development. In the present study, we examined the effect of prenatal exposure of Sig-1R antagonist on postnatal neuronal development and behavior in rat offspring. Pregnant Long-Evans rats were administered NE-100, a selective Sig-1R antagonist, once daily at a dose of 1.0 mg/kg（i.p.）from gestation day 14 to 20. Their young-adult（6 weeks old）male offspring were subjected to behavioral tests and morphological analysis using Golgi-Cox staining. Behavioral studies using the Barnes maze and Y-maze showed that prenatal NE-100 exposure was associated with cognitive impairment in the young-adult offspring. Furthermore, anxiety-like behavior, which was assessed using the elevated plus maze, increased in NE-100-exposed rats. In addition, Golgi-Cox staining revealed that NE-100 exposure disrupted the morphologies of dendrites resulting in decreased dendritic length and branching in granular neurons in the hippocampal DG and CA4 regions. These findings suggest that prenatal exposure to a Sig-1R antagonist is associated with impaired cognition and anxiety states through disruption of dendritic morphology in hippocampal neurons.

2P-53 Effects of oxytocin and analog, Lipo-oxytocin 1 on paternal behavior and social memory in CD38-/- mice Cherepanov Stanislav^1,2，Yokoyama Shigeru²，Shuto Satoshi³，Higashida Haruhiro² ¹United Graduate School of Child Development, Osaka University, Kanazawa University, Hamamatsu University School of Medicine, Chiba University and University of Fukui，²Research Center for Child Mental Development, Kanazawa University，³Faculty of Pharmaceutical Sciences, Hokkaido University Autism is neurodevelopmental disorder characterized by core deficits in sociability, repetitive behavior and restricted interests. One of the perspective fields of research of autism therapy is nanopeptide oxytocin（OT）. We have a lot of proofs about roles of oxytocin in the social interactions and social recognition. Now oxytocin is used intranasaly like an experimental drug for treatment patients with autism. To ensure such roles, an analog with long-lasting and effective blood-brain barrier penetration properties should have a benefit, like a therapeutic drug. To assess this, we synthesized a new oxytocin analog, lipo-oxytocin-1（LOT-1）, which conjugates two palmitoyl groups at the amino group of the cysteine and the phenolic hydroxyl group of the tyrosine in the OT molecule. In previous research, LOT-1 demonstrated long-effect on recovery of sociability in open field test in CD157-/- mice. Now, were investigated OT and LOT-1 on paternal behavior and social memory in CD38-/- mice. That mice are established model of autism symptoms. In parental behavior test CD38-/- male mice demonstrate low pups retrieving scores and high time for completely retrieve pups in the nest. In the case of treatment by OT or LOT-1, 30 minutes after injection CD38-/- demonstrate decreasing of complete time for retrieving and increase retrieving scores. Also after 24 hours of injection, mice treated by LOT-1 demonstrate increased parental behavior than at 30 minutes, while mice treated by OT demonstrate parameters are constant at 30 minutes and 24 hours.In the social memory test CD38-/- male mice demonstrate low level of social discrimination between familiar and novel mice. The OT and LOT-1 recover social memory in CD38-/- mice and the mice demonstrate similar pattern of behavior with wild-type. Finally, we show LOT-1 in vitro effects for oxytocin receptors. Together, these results suggest that LOT-1 has a functional advantage for recovery of social behavioral impairment.		2P-54 Expression of CD38 and TRPM2 in activated microglia and behavioral impact in mice lacking CD38 Suematsu Fumiya¹，Kojima Yuichiro¹，Higashida Haruhiro²，Noda Mami¹ ¹Lab Pathophysiol, Grad Sch Pharm, Kyushu Univ，²Dept Bas Res Social Recognition ＆ Memory, Res Cent for Chil Ment Devel, Kanazawa Univ CD38, whose molecular weight is 45 kDa, mainly expresses in immune cells and is involved with Ca²⁺ signaling through synthesis of cyclic ADP-ribose（cADPr）. Recently, it was reported CD38 is required for maintaining social behavior by regulating the secretion of oxytocin, a hypothalamus hormone being responsible for trust and generosity, in mice（Jin et al., 2007）. Previously we showed that expression of CD38 protein was detected in microglia and more expression of microglial CD38 was observed in the lipopolysaccharide（LPS）-injected mouse brain in vivo（Akimoto et al., 2013）. Using primary cultured mouse microglia, we also reported that application of LPS（100 ng/mL）, but not ATP（100 μM）, for 24 h up-regulated the expression of microglial CD38. In addition, knock-down of TRPM2, which colocalize with CD38, significantly up-regulated the expression of microglial CD38（Noda et al., 2014）. Therefore, the question to be answered is whether the CD38-related system in microglial cells is involved in autism as an immuno-inflammation factor. In the present study, we examined that the expression of microglial CD38 was not up-regulated either at 1, 2, 4, and 6 h after application of ATP（1 mM）, suggesting that up-regulation of CD38 is limited to LPS-induced inflammatory condition. We will report the regulation of expression of TRPM2 in CD38-deficient microglia by quantitative RT-PCR. In addition, the behavioral change in wild-type and CD38-knock out mice after application of LPS will be reported.

2P-55 Rheb activation disrupts spine synapse formation through accumulation of syntenin in tuberous sclerosis complex Yasuda Shin¹，Sugiura Hiroko¹，Katsurabayashi Shutaro²，Kawano Hiroyuki²，Endo Kentaro³，Takasaki Koutaro²，Iwasaki Katsunori²，Ichikawa Masumi³，Kobayashi Toshiyuki⁴，Hino Okio⁴，Yamagata Kanato¹ ¹Synaptic Plasticity Project, Tokyo Metropolitan Institute of Medical Science，²Department of Neuropharmacology, Faculty of Pharmaceutical Science, Fukuoka University，³Center of Basic Technology Research, Tokyo Metropolitan Institute of Medical Science，⁴Department of Pathology and Oncology, Juntendo University, School of Medicine Rheb is a small GTP-binding protein and its GTPase activity is activated by the complex of Tsc1 and Tsc2 whose mutations cause tuberous sclerosis complex（TSC）. We previously reported that cultured TSC neurons showed impaired spine synapse morphogenesis in an mTORC1-independent manner. Here we show that the PDZ protein syntenin preferentially binds to the GDP-bound form of Rheb. The levels of syntenin are significantly higher in TSC neurons than in wild-type neurons because the Rheb-GDP-syntenin complex is prone to proteasomal degradation. Accumulated syntenin in TSC neurons disrupts spine synapse formation through inhibition of the association between syndecan-2 and calcium/calmodulin-dependent serine protein kinase. Instead, syntenin enhances excitatory shaft synapse formation on dendrites by interacting with ephrinB3. Downregulation of syntenin in TSC neurons restores both spine and shaft synapse densities. These findings suggest that Rheb-syntenin signalling may be a novel therapeutic target for abnormalities in spine and shaft synapses in TSC neurons.		2P-56 Prenatal administration of valproic acid or tributyltin alters developmental transient of hippocampal excitability in juvenile rats Fueta Yukiko¹，Sekino Yuko²，Yoshida Sachiko³，Ueno Susumu⁴ ¹Department of Environmental Management and Control, University of Occupational and Environmental Health，²Division of Pharmacology, National Institute of Health Sciences，³Department of Environment and Life Science, Toyohashi University of Technology，⁴Department of Occupational Toxicology, University of Occupational and Environmental Health Valproic acid（VPA）is commonly used as an antiepileptic drug and a mood stabilizer, but is also known as a developmental neurotoxicant because animal models of autism spectrum disorder has been established by prenatal exposure to VPA. In this study we aimed to clarify with our electrophysiological approach whether prenatal VPA exposure affects early postnatal development of neuronal circuitry, before the appearance of neurobehavioral change in adolescent period. VPA was orally administered to the pregnant day15 Wistar rats with the concentrations of 300 mg/kg. On the days from PND 13 to 18, field potentials were recorded from the CA1 area of hippocampal slices obtained from the control and VPA groups to test development of the local circuits. Stimulation/response curves of field excitatory postsynaptic potential and those of population spike（PS）enhanced at PND14 and 15 in the VPA group, suggesting that developmental transient of hippocampal excitability may be hastened. On the other hand, similar approach was applied to juvenile rats which were prenatally exposed to 20 mg/kg of tributyltin（TBT）, known to be typical endocrine disrupter, and we found a significant decrease in PS amplitude at PND16 in TBT group, which seems to be a retardation of developmental transient of excitability. These results suggest that our electrophysiological approach using hippocampal slices obtained from juvenile rats may be useful to predict the appearance of developmental neurotoxicity after adolescent period.