一般ポスター

一般ポスター神経伝達物質・受容体-2

7月7日（金）　13:50-14:50　ポスター会場① 2P②-1 マウス小脳におけるグリシントランスポーター１(GlyT1)が担うアストロサイトへのグリシン取り込み機構の構築 Development of glycine uptake into astrocytes by glycine transporter 1 (GlyT1) in the mouse cerebellum. 清水　千草, 濱田　麻美, 下地　美寧, 高山　千利琉球大学大学院　医学研究科　分子解剖学講座 Chigusa Shimizu, Asami Hamada, Miya Shimoji, Chitoshi Takayama Dept. of Molecular Anatomy, School of Medicine, Univ. of the Ryukyus, Okinawa, Japan Glycine is an inhibitory transmitter in the cerebellum. Glycine released into synaptic cleft is removed by glycine transporter1 (GlyT1) localized in astrocytes and GlyT2 in nerve terminals. However, the development of the glycine uptake system into astrocytes in the cerebellum is still unclear.　We performed immunostaining for GlyT1 and examined its relationship with Purkinje cells and glycinergic terminals by using frozen sections of mouse cerebellum. The results showed that GlyT1 was expressed on short projections extending from nestin-positive fibers, a marker of radial glia, when the three layers of the cerebellar cortex is incomplete. At P0, GlyT1 was expressed around migrating Purkinje cells stained with calbindin. At P7, when the Purkinje cells were arranged in monolayer, GlyT1 was expressed surrounding all layer of neurons. After P21, GlyT1 expression became more on the axonal side of Purkinje cells and around dendrites spreading into the molecular layer. In the granular layer, GlyT2, a marker of glycinergic terminals, began to be expressed around the granule cells, and GlyT1 was expressed outside of them.　These results suggest that there is a glycine uptake　system into astrocytes before the formation of glycinergic terminals and that glycine uptake into astrocytes is observed throughout the cerebellar cortex regardless of the presence or absence of glycinergic ones.		7月7日（金）　13:50-14:50　ポスター会場① 2P②-2 DHAによる L-グルタミン酸トランスポーター電流増強作用にロイシン434は必要である Leucine 434 is essential for docosahexaenoic acid-induced augmentation of L-glutamate transporter function 高橋　華奈子¹, Luying Chen³, 佐山　美砂³, Mian Wu³, 林　真理子⁴, 入江　智彦², 大和田　智彦³, 佐藤　薫² 1. 東京大学大学院　薬学系研究科　天然物化学教室, 2. 国立医薬品食品衛生研究所　薬理部, 3. 東京大学大学院　薬学系研究科　薬化学教室, 4. 昭和女子大学　食健康科学部　管理栄養学科 Kanako Takahashi¹, Luying Chen³, Misa Sayama³, Mian Wu³, Mariko Kato Hayashi⁴, Tomohiko Irie², Tomohiko Ohwada³, kaoru sato² 1. Laboratory of Natural Products Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 2. Laboratory of Neuropharmacology, Division of Pharmacology, National Institute of Health Sciences,, 3. Laboratory of Organic and Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 4. Faculty of Food and Health Sciences, Department of Food Science and Nutrition, Showa Women University Astrocytic EAAT2 plays a role in removing the L-glutamate (L-Glu) from synaptic clefts to prevent excitotoxicity. Docosahexaenoic acid (DHA, 22:6) enhance synaptic transmission and their target molecules include EAATs. We studied the effect of DHA on EAAT2 and identify the key amino acid for DHA/EAAT2 interaction using two-electrode voltage clamp technique in Xenopus oocytes. DHA increased EAAT2 currents (A), but tended to decrease EAAT1, another astrocytic subtype, currents. Single mutation of Leu 434 to Ala suppressed the DHA-induced enhancement, while back mutation of EAAT1 Ala 435 (corresponding to EAAT2 Leu434) to Leu changed the effect from inhibition to enhancement. Docking analysis of EAAT2 and DHA suggested the most stable docking site is the lipid crevice, exists at the interface between trimerization domains and transport domains (B), close to the L-Glu and Na ⁺ binding site.

7月7日（金）　13:50-14:50　ポスター会場① 2P②-3 リン酸化情報基盤データベースKANPHOSが紐解くドーパミンシグナル伝達の分子基盤 Molecular basis of dopamine signaling unraveled by KANPHOS, a database of protein phosphorylation in the brain 坪井　大輔¹, 大塚　岳², 拓史　下村³, 船橋　靖広¹, 佐野　裕美^1,4, 永井　拓⁵, アナスタシオス　タジンゴニス⁶, 南部　篤⁴, 川口　康雄², 久保　義弘³, 貝淵　弘三¹ 1. 藤田医科大学　精神神経病態解明センター　細胞生物学部門, 2. 生理学研究所　大脳神経回路論研究部門, 3. 生理学研究所　神経機能素子研究部門, 4. 生理学研究所　生体システム研究部門, 5. 藤田医科大学　精神神経病態解明センター　行動薬理学部門, 6. 米国コネティカット大学　神経生理学講座 DAISUKE TSUBOI¹, Takeshi Otsuka², Takushi Shimomura³, Yasuhiro Funahashi¹, Hiromi Sano^1,4, Taku Nagai⁵, Tzingounis Anastasios V⁶, Atsushi Nambu⁴, yasuo Kawaguchi², Yoshihiro Kubo³, Kozo Kaibuchi¹ 1. Division of Cell Biology, International Center for Brain Science, Fujita Health University, 2. Division of Cerebral Circuitry, National Institute for Physiological Sciences, 3. Division of Biophysics and Neurobiology, National Institute for Physiological Sciences, 4. Division of System Neurophysiology, National Institute for Physiological Sciences, 5. Division of Behavioral Neuropharmacology, International Center for Brain Science, Fujita Health University, 6. Department of Physiology and Neurobiology, University of Connecticut Protein phosphorylation plays a critical role in various intracellular signaling pathways and physiological functions controlled by neuromodulators, including dopamine. We built "KANPHOS", a novel online database for protein phosphorylation, to understand the links of phosphorylation to upstream signaling events. Dysregulation of dopamine signaling in the nucleus accumbens (NAc), a part of the basal ganglia in the brain, has been implicated in psychiatric disorders. We have previously reported that dopamine increases excitability and firing rates of D1 receptor (D1R)-expressing medium spiny neurons (MSNs) in the nucleus accumbens (NAc) via the PKA/Rap1/ERK pathway to promote reward behavior. However, the mechanism by which ERK acts downstream of dopamine signaling regulates MSN excitability. Substrate search with KANPHOS identified an ERK phospho-substrate known as potassium voltage-gated channel subfamily Q member 2 (KCNQ2). We found that agonist activation of D1R receptors inhibited KCNQ-dependent currents and led to increased D1R MSN firing rates in mouse NAc slices. MSN firing was halted upon inhibition of ERK. Further analysis confirmed that direct phosphorylation of KCNQ2 by ERK occurs through activation of the dopamine signaling pathway for reward behavior. Hence, D1R-ERK signaling controls MSN excitability via KCNQ2 phosphorylation to regulate reward behavior.		7月7日（金）　13:50-14:50　ポスター会場① 2P②-4 マウス脳海馬CA3領域への特異的なPP5過剰発現は不安様行動を示す Specific PP5 overexpression in the CA3 region of the mouse brain hippocampus is associated with anxiety-like behavior 宇野　恭介, 河原井　康介, 上田　舞那水, 金城　俊彦, 倉本　展行摂南大学　薬　機能形態学 Kyosuke Uno, Kosuke Kawarai, Manami Ueda, Toshihiko Kinjyo, Nobuyuki Kuramoto Lab. Mol. Pharmacol., Fac of Pharm. Sci., Setsunan Univ. We have explored the possibility that protein phosphatase 5 (PP5), together with PP1 and PP2A, promotes desensitization of GABAB receptors and attenuates receptor action by dephosphorylating the 892nd serine residue of mouse GABAB receptors. In this study, we investigated the possibility that dephosphorylation of GABAB receptors in the hippocampal CA3 region affects the behavior of mice.The PP5 gene was cloned in an adeno-associated viral vector (AAV). Mice in which the PP5 gene was overexpressed in the hippocampal CA3 region (PP5 mice) using the created AAV or mice in which AAV containing only the GFP gene was introduced as a control group (GFP mice) were created, and the effect on mouse behavior was analyzed.In the Open field test, PP5 mice tended to stay in the center area longer than GFP mice. In an elevated cross-maze test, PP5 mice tended to stay longer in open zones than GFP mice. In the Y-maze test, PP5 mice showed a decreasing trend in alternation behavior compared to GFP mice. In the novel object search test, a significant increase in the time of interest in the novel object was observed in GFP mice, but in PP5 mice there was no significant difference in the time of interest in the two objects.The behavioral experiments show that overexpression of PP5 in the CA3 region may affect mice to reduce anxiety-like behavior and cognitive function.

7月7日（金）　13:50-14:50　ポスター会場① 2P②-5 非ステロイド性抗炎症薬Flurbiprofenは、分子シャペロン誘導剤として働き、セロトニントランスポーターの機能を制御する。 The nonsteroidal anti-inflammatory drug flurbiprofen regulates serotonin transporter function. 酒井　規雄, 平川　明樹, 田口　慧, 村川　青矢, 浅野　昌也, 野口　颯真, 吉川　慧, 原田　佳奈, 秀　和泉, 田中　茂広島大学　医系科学研究科　神経薬理学 Norio Sakai, Haruki Hirakawa, Kei Taguchi, Seiya Murakawa, Masaya Asano, Soma Noguchi, Satoru Kikkawa, Kana Harada, Izumi Hide, Shigeru Tanaka Dept of Pharmacol Neurosci, Grad Sch Biomed Sci, Hiroshima Univ Serotonin transporter (SERT) function is regulated by membrane trafficking. We have been searching for drugs that promote membrane trafficking of SERT. Flurbiprofen has been reported to exhibit chemical chaperone activity. Therefore, we investigated the effects of Flurbiprofen on SERT function.We used COS-7 cells transiently expressing wild-type (WT) SERT or a C-terminal-deleted mutant of SERT (SERTΔCT), a misfolded protein, to examine the effects of flurbiprofen on serotonin uptake activity, protein expression level, and glycosylation state of SERT. Treatment of WT-SERT-expressing cells with Flurbiprofen (＞0.5 mM) for 24 hours decreased the immaturely-glycosylated SERT and increased the maturely- glycosylated SERT, suggesting that flurbiprofen modulates SERT function by promoting SERT membrane trafficking. Studies with enantiomers of flurbiprofen revealed that these effects of flurbiprofen are not mediated by cyclooxygenase inhibition. We investigated whether Flurbiprofen could ameliorate SERTΔCT-induced ER stress by evaluating the level of the GRP78/BiP, a marker of ER stress. Interestingly, we found that flurbiprofen induced GRP78/BiP expression only under ER stress conditions, but not under steady-state conditions. These results suggest that flurbiprofen may also function as an inducer of molecular chaperones in addition to as a chemical chaperone.