Poster Sessions

Poster Sessions 神経ネットワーク、その他

1P-46 Autism-like behavioral abnormalities in N-ethylmaleimide - sensitive factor knockout mice Min Jue Xie^1,2,3，Keiko Iwata^1,2,3，Yugo Fukazawa^1,4，Hideo Matsuzaki^1,3 ¹Div Dev Mental Func, Res Center Child Mental Dev, Univ of Fukui，²Life Sci lnnovation Center, Univ of Fukui，³United Grad Sch Child Dev, Osaka Univ, Kanazawa Univ, Hamamatsu Univ Sch Med, Chiba Univ, Univ of Fukui，⁴Div brain struc func, Dept Morphol Physiol Sci, Grad Sch Med Sci, Univ of Fukui, Fukui, Japan Autism, characterized by profound impairment in social interactions and communicative skills, is the most common neurodevelopmental disorder. Many studies on the mechanisms of autism have focused on the serotonergic system but its underlying molecular mechanisms remain controversial. In our previous report, we reidentified N-ethylmaleimide-sensitive factor (NSF) as new serotonin transporter binding protein, which expression reduced in postmortem brains and lymphocytes of autistic subjects (Iwata et al 2014). In this study, we generated the NSF+/- mice and investigated their behavioral phenotypes. As previous report has already shown that NSF is necessary for AMPA receptors location in the synapse, we examined AMPA receptors location in the synapse of the NSF+/- mice by using freeze-fractured replica-immunolabeling study at first, and revealed the significant decrease in postsynaptic expression of AMPA receptors in their brain. Then, we assessed the social interaction behaviors using the three-chambered task and found that the spending time near the chamber containing the novel mouse were significantly reduced in the NSF+/- mice, compared with wild mice. It is suggesting that cellular trafficking turbulence of synaptic molecules by lacking NSF gene might be related to the pathophysiology of autistic behavior.		1P-47 The mitochondrial dysfunction due to the accumulation of mtDNA deletions in the brain causes behavioral abnormalities in heterozygous Polg mutant mice Satoshi Fuke^1,2，Yasufumi Shigeyoshi³，Jin Nakatani⁴，Saori Yoshida⁵，Ken-ichi Mukaisyo⁵，Mie Kubota-Sakashita²，Natsu Koyama¹，Hiroyuki Sugihara⁵，Seiji Hitoshi¹，Tadafumi Kato² ¹Department of Integrative Physiology, Shiga University of Medical Science，²Laboratory for Molecular Dynamics of Mental Disorders, RIKEN BSI，³Department of Anatomy and Neurobiology, Kinki University School of Medicine，⁴Biomedical Magnetic Resonance Science Unit, Molecular Neuroscience Research Center, Shiga University of Medical Science，⁵Department of Pathology, Division of Molecular and Diagnostic Pathology, Shiga University of Medical Science Chronic progressive external ophthalmoplegia (CPEO) with accumulations of multiple mitochondrial DNA (mtDNA) deletions in the brain and muscles often accompanied with mood disorders including recurrent depression and bipolar disorder. However, a causal relationship between the mutant mtDNA in the brain and psychiatric symptoms remains unclear. We previously reported that the age-dependent accumulations of mtDNA deletions were accelerated in the brain and muscles of heterozygous knock-in mice with mutant gene encoding the proofreading-deficient Polg (Polg^D257A), which is one of causative gene of CPEO (Fuke et al., Ann Clin Transl Neurol. 2014). Here, we demonstrate that Polg^+/D257A mice showed the altered behavioral rhythm. Furthermore, we observed that the accumulation of mtDNA deletions in a number of subregions in the brain was significantly higher in Polg^+/D257A mice than in Polg^+/+ mice, and found the significant subregional variation of the fold change. In particular, mtDNA deletions were markedly increased in prelimbic, infralimbic cortices and the nucleus accumbens, which have been implicated in mood disorders. Immunohistochemistry staining of mitochondrial proteins, the mtDNA-encoded MTCOI and the nuclear-encoded SDHA, revealed the presence of COX-deficient cells in prelimbic cortex of Polg^+/D257A mice. Proton magnetic resonance spectroscopic investigations in Polg^+/D257A mice showed a tendency of elevated lactate levels in regions containing prelimbic/infralimbic cortices and the nucleus accumbens. These results suggest that local mitochondrial dysfunctions due to the accumulation of mtDNA deletions can induce neuropsychiatric symptoms.

1P-48 Selective serotonin reuptake inhibitors with affinity for σ₁ receptors exert anti-anhedonic effect in picrotoxin-induced model of anxiety/depression Yukio Ago¹，Shigeru Hasebe^1,2，Momoko Higuchi¹，Hitoshi Hashimoto^1,3,4，Kazuhiro Takuma^1,2,3，Toshio Matsuda⁵ ¹Lab. Mol. Neuropharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ.，²Dept. Pharmacol., Grad. Sch. Dent., Osaka Univ.，³United Grad. Sch. Child Dev., Osaka Univ.，⁴Inst. Datability Sci., Osaka Univ.，⁵Lab. Med. Pharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ. Selective serotonin (5-HT) reuptake inhibitors (SSRIs) are first-line agents in the treatment of mood and anxiety disorders. The SSRIs fluvoxamine and S-(+)-fluoxetine have high affinity for the σ₁ receptor, while paroxetine does not. The σ₁ receptor is involved in psychiatric disorders, but it remains unclear whether the effect on the receptor plays a key role in the pharmacological effects of SSRIs. We previously found that fluvoxamine, but not paroxetine, activates prefrontal dopaminergic system under the conditions of decreased brain GABA_A receptor function via combined activation of 5-HT_1A and σ₁ receptors. Here we aimed to clarify whether SSRIs with an affinity for σ₁ receptors have anti-anhedonic effects. Male mice were treated with picrotoxin to decrease GABA_A receptor function. Their anhedonic behavior was measured using the female encounter test, a novel method for measuring sexually motivated behavior. Picrotoxin decreased the preference for female encounter and attenuated the female encounter-induced increase in accumbal c-Fos expression. Picrotoxin-induced anhedonia was ameliorated by fluvoxamine and S-(+)-fluoxetine, but not by paroxetine. The effect of fluvoxamine was blocked by a 5-HT_1A or a σ₁ receptor antagonist, and co-administration of the σ₁ receptor agonist (+)-SKF-10047 and the 5-HT_1A receptor agonist osemozotan mimicked the effect of fluvoxamine. The effect of fluvoxamine was also blocked by a dopamine-D_2/3 receptor antagonist, and methylphenidate, an activator of the prefrontal dopamine system, ameliorated picrotoxin-induced anhedonia. These findings suggest that SSRIs with agonistic activity at σ₁ receptors exert anti-anhedonic effect under the conditions of decreased GABA_A receptor function such as anxiety and depression.		1P-49 Cortical interleukin-1 facilitates learning the spatial memory task in the water maze in young mice Takako Takemiya¹，Kanato Yamagata²，Yoichiro Iwakura³，Marumi Kawakami¹ ¹Med Res Insti, Tokyo Women’s Med Univ，²Synaptic Plasticity Proj, Tokyo Metro Insti of Med Sci，³Insti for Biomed Sci, Tokyo Univ of Sci Interleukin-1 (IL-1) is produced in the periphery and glia and neurons in the brain. The type I IL-1 receptor (IL-1r1) is primarily responsible for transmitting the inflammatory effects of IL-1 and mediates several biological functions by binding to either IL-1α or IL-1β. IL-1β activation is associated with hippocampus-dependent memory task. Although IL-1β impairs spatial memory under certain pathophysiological conditions, IL-1 β may be required for the normal physiological regulation of hippocampal plasticity and memory. In addition, brain IL-1β levels are thought to be increased in the hippocampus in an age-dependent manner. Therefore, we hypothesized that hippocampal IL-1β signaling has a beneficial effect on spatial learning and memory in young mice via IL-1r1 that is diminished in adults. We investigated the performance of young (3-month-old) and adult (6-month-old) wild-type mice (wt), IL-1β  knockout mice (IL-1βko) and IL-1r1 knockout mice (IL-1r1ko) in learning a spatial memory task with fixed platform (PF) in the water maze (WM) and measured the levels of IL-1β and IL-1α in the hippocampus and cortex with IL-1β immunohistochemistry using adult and young mice brain tissue. Learning was significantly impaired in training trials of the spatial memory task in the WM in young IL-1βko and young IL-1r1ko but not in adult IL-1βko or adult IL-1r1ko mice. This suggests that IL-1β has a crucial role in learning in the spatial memory. Furthermore, the cortical concentration of IL-1β and IL-1α was significantly increased in adult wt mice compared to young wt mice, and IL-1β was increased in neurons in the cortex in adult mouse. Our results suggest that cortical IL-1 accelerates learning the hippocampal spatial memory task in the WM in young mice.

1P-50 Prepulse inhibition in Drosophila reveals possible candidate genes that have functions in sensory gating and relate to the psychiatric disorder. Takako Morimoto，Yutaro Matsumoto，Koki Takei，Kazuya Shimizu，Junji Yamauchi Lab of Neuroscience and Neurology, Sch of Life Sci, Tokyo Univ of Pharm and Life Sci The neural mechanisms of the psychiatric disorder like autism spectrum disorder and schizophrenia has been intensively studied and a number of genes have been identified as candidates that cause the disorder. However, the inclusive understanding of relation between the gene and the function of neural system has not been clarified yet. To solve this problem, studies on a model organism, Drosophila, may serve as a powerful tool because of its small size of neural cell number and established genetic methods. In the last meeting, we reported a prepulse inhibition (PPI) phenomenon by using Drosophila larval hearing. PPI is a neurological phenomenon found in humans and other organisms and using for the diagnostic test of schizophrenia as well as autism. A weaker prestimulus (prepulse) inhibits the reaction to a subsequent strong startling stimulus (pulse). Our finding is the first report of PPI in Drosophila. Here, we have examined the PPI of two mutants in which their responsible genes are supposed to relate to the psychiatric disorder. One is fmr1, which is a responsible gene of fragile x syndrome. Another is centaurin. Centaurin family members commonly have domains, such as GTPase, PH, ArfGAP and ANK domains. In addition, it has been reported that one of centaurin family members is included in the chromosomal deletion region found in autistic patients. Further, we showed its potential function as a negative regulator of neurotransmitter release (Homma, et al, 2014). We found that these mutants showed reduced PPI. In fmr1, they also showed the defective in the pulse-induced startling response itself, but not in centaurin. We are further examining the genetic interaction of these proteins and will discuss more.		1P-51 The interaction between respiratory rhythm and body movement in the Parabrachial nucleus in the developmental stage Akiko Arata¹，Chiaki Uchida¹，Hirotaka Ooka¹，Suguru Iwano^1,2，Toshiki Noma^1,2，Sotatsu Tonomura^1,3，Akira Tamaki² ¹Dept. of Physiol. Hyogo Coll. of Med.，²Hyogo Univ. of Helth Science, Physical Therapy for Internal Disorders，³Dept. of Anatomy, Hyogo Coll. of Med. Parabrachial nucleus (PB) is thought to be correlated with the autonomic function and the relationship of sensory-motor integration. The PB is also known as a respiratory modulating center and the PB plays a crucial role in the inspiratory-expiratory (I-E) phase switching. In this study, we focused to examine how the PB participates in the relationship between respiration and body movement using pons-medulla-spinal cord preparations obtained from 0 to 4 days old rats. First, the effects of C8/L4 dorsal root stimulation on C4 ventral root inspiratory activity were examined. The electrical stimulation of dorsal root of C8 (brachial) level or L4 (pedal) level induced respiratory rhythm resetting. We describe the respiratory neurons in the dorsal pons and their interacting the respiration and body movement. We found I-E neurons which was major of the recorded neuron in the PB using whole cell patch clamp method. When PB was stimulated electrically, respiratory rate was increased. I-E neurons might make respiratory cycle faster using inspiratory-expiratory phase switching system. The body movement remained activity even if medulla was removed, but the frequency significantly decreased. It was suggested that the body movement generated in spinal cord, but might receive some excitatory input from the medulla. The projection to PB by the dorsal root stimulus of C8 was confirmed using optical imaging. The C8 (or L4) dorsal root stimulation induced respiratory excitation via PB. These results suggested that the sense of hand and foot is projected on PB, then I-E neuron in the PB activated respiratory rhythm. In the relationship between respiratory activity and body movement, the entrainment of both rhythmic activities were induced by PB as a sensory-motor coordination.

1P-52 Enhanced locomotor activity and impaired prepulse inhibition induced by presynaptic cytomatrix protein Piccolo knockdown in the amygdala of mice Kohei Hamatani¹，Yoshiaki Miyamoto¹，Yuki Oketani¹，Shin-iti Muramatsu^2,3，Kyosuke Uno¹，Atsumi Nitta¹ ¹Department of Pharmaceutical Therapy & Neuropharmacology, Faculty of Pharmaceutical Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama，²Division of Neurology, Department of Medicine, Jichi Medial University，³Center for Gene &Cell therapy, The Institute of Medical Science, The University of Tokyo Schizophrenia is defined as a mental disorder that causes one to suffer positive symptom, negative symptom and cognitive dysfunction. The multiple risk factors including genetic predisposition are involved in the onset of schizophrenia. A previous report revealed elevated PCLO mRNA in the amygdala of the patients with schizophrenia. In contrast, the same research group reported that the expression levels of PCLO mRNA increased in SH-SY5Y cells after treatment with atypical antipsychotic drugs, but not typical antipsychotic drugs. PCLO encodes presynaptic cytomatrix protein Piccolo. In this study, we investigated the causal relationship between Piccolo expression and schizophrenia using mice. Like the aforementioned in vitro study, the protein expression levels of Piccolo increased in the amygdala of mice repeatedly treated with risperidone, an atypical antipsychotic drug, but not haloperidol, a typical antipsychotic drug. These findings suggest that elevated PCLO mRNA in the postmortem brain of schizophrenia patients is due to the therapeutic efficacy, rather than the pathogenesis of schizophrenia. Accordingly, we investigated the phenotypical changes in mice with Piccolo knockdown in the amygdala. To generate animals with reduced Piccolo expression, we injected AAV-miPCLO vector into the amygdala of the 8-week-old mice. Four weeks later, the Piccolo knockdown mice showed hyperlocomotion in the novel environment and impaired acoustic prepulse inhibition. These observations suggest that dysfunction of Piccolo in the amygdala associates with the schizophrenia-like behavior in mice. The investigation of the cognitive function and depressive behaviors is necessary in the Piccolo knockdown mice to evaluate as the novel animal model for schizophrenia.		1P-53 Links among molecular clock, protein modification and adaptive protection systems to support 24hr-lifestyle Teruya Tamaru¹，Genki Kawamura²，Hikari Yoshitane³，Yoshitaka Fukada³，Takeaki Ozawa²，Ken Takamatsu¹ ¹Dept Physiology, Toho Univ Sch Med，²Dept Chemistry, Sch Sci, Univ Tokyo，³Dept Biological Sciencence, Sch Sci, Univ Tokyo In modern society without clear day-night boundary, humans are often exposed to lifestyles departing beyond adaptation capacity of molecular clock program, which was acquired after a long evolution. Thereby, it gradually undermines our health based on regular 24hr-lifestyle, and occasionally causes diseases. Cellular molecular clocks, which work through the negative feedback loop composed with clock genes/ proteins, are timekeepers for various physiological functions by brain SCN (central clock) -orchestrating temporal harmony of peripheral clocks of tissues/ organs in whole body. In this topic, we will present/ discuss on; 1) molecular clock function at single cellular level and cellular clock synchronization, 2) protein modifications (phosphorylation, ubiquitination, acetylation, etc.), interaction, degradation and nucleocytoplasmic shuttling processes, which are critical in the linking between clock and various adaptive protection systems, such as heat shock and anti-oncogenic system, etc.

1P-54 Effects of SHATI/NAT8L overexpression in the striatum on social stress sensitivity in mice Hajime Miyanishi¹，Kyosuke Uno¹，Toh Miyazaki¹，Kengo Sodeyama¹，Toshiyuki Fujiwara¹，Shin-ichi Muramatsu^2,3，Yoshiaki Miyamoto¹，Atsumi Nitta¹ ¹Department of Pharmaceutical Therapy and Neuropharmacology, Faculty of Pharmaceutical Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama，²Division of Neurology, Jichi Medical University，³Center for Gene & Cell Therapy, The Institute of Medical Science, The University of Tokyo The number of depression patients is increasing. The mechanism of depression onsets has not been completely clarified. It is reported that the SHATI/NAT8L-overexpressed mice exhibited depression like-behaviors. SHAT/NAT8L synthesize N-asetyl-aspartate (NAA) from L-aspartate and acetyl-CoA. NAA is converted to N-acetylaspartylglutamate (NAAG), an agonist of mGluR3, by condensation with glutamate. In this study, we investigated the relationship between SHATI/NAT8L and depression model mice exposed to social defeat stress.C57BL/6J mice after exposure to repeated social stress (exposure to ICR mice for 10min/day, 10days) showed depression-like behaviors, decreasing social interaction and sucrose preference. Next, we generated the SHATI/NAT8L-overexpressed mice by injecting an adeno-associated virus into the dorsal striatum. The SHATI/NAT8L-overexpressed mice exhibited decreased social interaction and sucrose preference after the subthreshold social defeat stress (exposure to ICR mice for 5min, 3times) which showed normal behaviors in the control mice. These results suggest that SHATI/NAT8L was related to social stress sensitivity. The depression-like behavior was restored by the selective serotonin reuptake inhibitor fluvoxamine and the selective mGluR3 antagonist LY341495.Taken together, SHATI/NAT8L has an important role in social stress sensitivity which is related to depression onsets by regulating serotonergic neuronal system. Striatal SHATI/NAT8L might be a new target for medical tools addressing depression. Further investigation is required to clarify the detailed relationship between stress sensitivity and SHATI/NAT8L.		1P-55 Regulation of lipid droplets by Mycobacterium leprae infection in Schwann cells Masumi Endoh Dept Mycobacteriol, Leprosy Research Center, NIID Leprosy is disease caused by infection of Mycobacterium leprae (ML). ML shows high affinity to Schwann cells (SCs), and serious peripheral neuropathy thereby occurs. The mechanisms responsible for peripheral neuropathy in leprosy need further elucidation. However, it is demonstrated that a lipid droplet (LD) is formed after ML infection in SCs. And it is speculated that the accumulation of the LD has an influence on the peripheral nerve repair mechanisms such as the remyelination by adjusting the lipid metabolism of SCs. The present study investigated the LD formation in ML-infected SCs. The immortalized Schwann cell line derived from rat sciatic nerve were used. ML prepared from the footpads of athymic nu/nu mice which maintained in our laboratory. SCs were incubated with ML at a multiplicity of infection (MOI) of 10:1 to 100:1. Alternatively, Schwann cells were stimulated with other acid-fast bacillus or latex beads. After 24 hours to 72 hours incubation, cells were fixed in 4% PFA. For fluorescent LD-labelling, cells were incubated with BODIPY493/503 dye. We evaluated the intracellular localization of the LD with a confocal laser scanning microscope. Also, we evaluated the lipids cumulative dose by used flow cytometer in some cultured SCs. The formation of the LD was found in the cytoplasm of SCs by infection of ML, but not found in circumference of the cell membrane and in the nucleus. The adipose cumulative dose depended on MOI and the duration of infection. The LD formation in SCs was observed only at ML infection and was not found at other acid-fast bacillus infection such as the BCG. Our data indicated that host cell lipid metabolism to ML infection are intimately related in SCs, and contributing to bacterial survival in the nerve and peripheral neuropathy.

1P-56 Involvement of BRINP1 in regulation of neural excitation Miwako Kobayashi¹，Shota Yamasaki¹，Yuichi Hayashi¹，Kono Yasuhiro¹，Kenshi Takechi^2,3，Akihiro Tanaka²，Hiroaki Araki²，Ichiro Matsuoka¹ ¹Col. of Pharm. Sci., Matsuyama Univ.，²Division of Pharmacy, Ehime Univ. Hospital，³Clinical Trial Ctr. for Developmental Therapeutics, Tokushima Univ. Hospital BRINP (BMP/RA-inducible Neural Specific Protein)-1, 2, 3 are family genes that are specifically expressed in the nervous system. Among the three family members, BRINP1 is expressed abundantly in various brain regions, and its expression is further up-regulated in dentate gyrus (DG) by administration of excitatory substances. Furthermore, BRINP1-KO mouse showed abnormal behaviors that are relevant to symptoms of schizophrenia and/or ADHD. In this study, to clarify the physiological role of BRINP1 in epileptic conditions, we examined c-Fos expression of DG neurons in pentylenetetrazol (PTZ) induced kindled BRINP1-KO mouse. We compared the number of c-Fos immunoreactive neurons between non-kindled and kindled mice at either steady state or post-seizure condition. At steady state, BRINP1 negatively regulated c-Fos expression in DG of kindled mouse. On the other hand, BRINP1 positively regulated c-Fos expression in DG of non-kindled mouse after severe epileptic seizure. Furthermore we examined the number of Parvalbumin (PV) immunoreactive neurons in prefrontal cortex in BRINP1-KO mouse. The number of PV neurons in BRINP1-KO mouse were half of those in WT mouse in both medial orbital cortex and prelimbic cortex, those are responsible for social behavior. This reduced PV immunoreactivities in prefrontal cortex in BRINP1-KO mouse is consistent with reduced PV-mRNA levels in human schizophrenic reported previously. These results suggest that BRINP1 regulates neuronal excitation in different ways; regulating c-Fos expression in excitatory neurons and regulating population of PV-expressing inhibitory neurons. Loss of these BRINP1 functions may disturb higher brain function and consequently lead to appearance of abnormal behaviors.		1P-57 Study of Teneurin-4 function to establish the bipolar disease mice model using CRISPR-Cas9 system Masaya Miyashita^1,2，Kyousuke Uno³，Fumitaka Nakano^1,2，Hirofumi Nishizono⁴，Shin-ichi Muramatsu^5,6，Yoshiaki Miyamoto³，Atsumi Nitta³ ¹Faculty of Pharmaceutical Science, University of Toyama, Toyama, Japan，²Department of Pharmaceutical Therapy and Neuropharmacology, Faculty of Pharmaceutical Science, University of Toyama, Toyama, JAPAN，³Department of Pharmaceutical Therapy and Neuropharmacology, Faculty of Pharmaceutical Science, Graduate School of Medicine and Pharmaceutical Science, University of Toyama, Toyama, JAPAN，⁴Life Science Research Center, University of Toyama, Toyama, JAPAN，⁵Division of Neurology, Jichi Medical University, Shimotsuke, Japan，⁶Center for Gene & Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Recently, the number of the patients with bipolar disorder is increasing and becoming one of the most seriously problems. It is too difficult to develop the remedy of bipolar disorder because of its characterization which repeat the depression and aggression. There are no agents to cure bipolar disorder at the present. In these years, genome-wide association study (GWAS) reported that single nucleotide polymorphism (SNP) at Odz4 which encodes Teneurin-4 was associated with bipolar disorder. Teneurin-4 is not revealed the detail of the function and role in the brain. In this study we investigated the function of Odz4 /Teneurin-4 to find neuronal mechanism of cause of bipolar disorder.Teneurin-4 was detected in the various brain regions of the normal mice by Western blotting, such as olfactory bulb, nucleus accumbens and hippocampus. Next, we designed three guide RNA to cut a part of exon of Odz4, and assessed cleavage ability by transfection of gRNA and Cas9 protein in the Neuro-2A cells. One of these gRNA could cut DNA and insert some mutations. To make Teneurin-4 knock-out Neuro-2A cell, we introduced vector containing Cas9-gRNA-GFP into Neuro-2A cells, and try to isolate a mutated cell line. We used these cells to investigate what kind of mutation was there or how weak Teneurin-4 expressed in mRNA level and protein content. To make Teneurin-4 knock-out mouse, we also did introduced gRNA and Cas9 protein into fertilized mice eggs by electroporation methods. We got two types of mutation in a F1 mouse.To establish Teneurin-4 knock-out mouse and Neuro-2A cell lines, we will able to investigate the roles of Teneurin-4, and supply a bipolar disorder mice model. They are expected to help developments for a new medical tool of bipolar disorder.