Pathophysiological analysis of epilepsy with focusing on the synaptic release mechanism

• Project/Area Number: 26460111
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pharmacology in pharmacy
• Research Institution: Osaka University of Pharmaceutical Sciences
• Principal Investigator: Yukihiro Ohno 大阪薬科大学, 薬学部, 教授(移行) (00432534)
• Co-Investigator(Renkei-kenkyūsha): KAWAI YOSHIKO 大阪薬科大学, 薬学部, 講師 (50268299) ; SHIMIZU SAKI 大阪薬科大学, 薬学部, 助手 (00630815)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: てんかん / シナプス分泌 / シナプス小胞蛋白SV2A / GABA遊離 / シナプトタグミン１ / てんかん原性 / キンドリング / 神経伝達物質 / GABA / 大脳辺縁系 / シナプス / シナプス小胞タンパク2A （SV2A) / 開口分泌 / グルタミン酸 / シナプス小胞蛋白2A (SV2A) / 動物モデル

Outline of Final Research Achievements

Synaptic vesicle glycoprotein 2A (SV2A) mediates action potential-dependent neurotransmitter release. To investigate the role of SV2A in regulating epileptogenesis, we generated a novel rat model carrying a Sv2a-targeted missense mutation (L174Q) and analyzed its seizure susceptibilities. The Sv2a mutant rats are highly susceptible to pentylenetetrazole (PTZ) seizure. Kindling with repeated PTZ treatments or amygdala stimulation was also markedly facilitated by the Sv2aL174Q mutation. In vivo microdialysis studies revealed that the Sv2aL174Q mutation reduced depolarization-induced GABA, but not glutamate, release in the limbic regions. Furthermore, the Sv2aL174Q mutation reduced the synaptotagmin1 (Syt1) level. The present results demonstrate that dysfunction of SV2A due to the Sv2aL174Q mutation impairs the synaptic GABA release by reducing the Syt1 level and facilitates the kindling development, illustrating the crucial role of SV2A-GABA system in regulating epileptogenesis.

Research Products

• [Journal Article] Therapeutic role of synaptic vesicle glycoprotein 2A (SV2A) in modulating epileptogenesis. (2017)
  • Author(s): Y. Ohno, K. Tokudome
  • Journal Title: CNS Neurol Disord Drug Targets Volume: 16 Issue: 4 Pages: 463-471
  • DOI: 10.2174/1871527316666170404115027
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Nicotine elicits convulsive seizures by activating amygdalar neurons. (2017)
  • Author(s): Iha HA, Kunisawa N, Shimizu S, Tokudome K, Mukai T, Kinboshi M, Ikeda A, Ito H, Serikawa T, Ohno Y
  • Journal Title: Frontiers in Pharmacology Volume: 8 Pages: 57-57
  • DOI: 10.3389/fphar.2017.00057
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Missense Mutation of the Gene Encoding Synaptic Vesicle Glycoprotein 2A (SV2A) Confers Seizure Susceptibility by Disrupting Amygdalar Synaptic GABA Release. (2016)
  • Author(s): Tokudome K, Okumura T, Terada R, Shimizu S, Kunisawa N, Mashimo T, Serikawa T, Sasa M, Ohno Y.
  • Journal Title: Frontiers in Pharmacology Volume: 7 Pages: 210-210
  • DOI: 10.3389/fphar.2016.00210
  • NAID: 120006331984
  • Peer Reviewed / Open Access
• [Journal Article] Synaptic vesicle glycoprotein 2A (SV2A) regulates kindling epileptogenesis via GABAergic neurotransmission. (2016)
  • Author(s): Tokudome K, Okumura T, Shimizu S, Mashimo T, Takizawa A, Serikawa T, Terada R, Ishihara S, Kunisawa N, Sasa M, Ohno Y.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 2742-2742
  • DOI: 10.1038/srep27420
  • NAID: 120006349333
  • Peer Reviewed / Open Access
• [Journal Article] Nicotine evokes kinetic tremor by activating the inferior olive via α7 nicotinic acetylcholine receptors. (2016)
  • Author(s): N. Kunisawa, H. A. Iha, S. Shimizu, K. Tokudome, T. Mukai, M. Kinboshi, T. Serikawa, Y. Ohno: Nicotine evokes kinetic tremor by activating the inferior olive via α7 nicotinic acetyl
  • Journal Title: Behav. Brain Res. Volume: 314 Pages: 173-180
  • DOI: 10.1016/j.bbr.2016.08.013
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Immunohistochemical analysis of Fos protein expression for exploring brain regions related to central nervous system disorders and drug actions. (2016)
  • Author(s): H.A. Iha, N. Kunisawa, K. Tokudome, T. Mukai, M. Kinboshi, S. Shimizu, Y. Ohno
  • Journal Title: Neuromethods Volume: 121 Pages: 389-408
  • DOI: 10.1007/978-1-4939-6490-1_17
  • ISBN: 9781493964888, 9781493964901
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] New insight into the therapeutic role of the serotonergic system in Parkinson’s disease (2015)
  • Author(s): Y. Ohno, S. Shimizu, K. Tokudome, N. Kunisawa, M. Sasa
  • Journal Title: Progress in Neurobiology Volume: 134 Pages: 104-121
  • DOI: 10.1016/j.pneurobio.2015.09.005
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Role of synaptic vesicle glycoprotein 2A (SV2A) in modulating epileptogenesis. (2017)
  • Author(s): Yukihiro Ohno
  • Organizer: 5th Congress of AsCNP
  • Place of Presentation: Indonesia (Bali)
  • Year and Date: 2017-04-27
  • Int'l Joint Research / Invited
• [Presentation] Function of Synaptic Vesicle Glycoprotein 2A as a Novel Therapeutic Target for Epilepsy (2017)
  • Author(s): Yukihiro Ohno
  • Organizer: 第90回日本薬理学会総会 （シンポジウム：てんかん研究の最前線－新たな治療薬を求めて）
  • Place of Presentation: 長崎
  • Year and Date: 2017-03-15
  • Invited
• [Presentation] Acute nicotine causes convulsive seizures by activating amygdala neurons, primarily through α7nACh receptors (2016)
  • Author(s): Higor A. Iha、國澤直史、清水佐紀、熱田侑大、野原孝紀、吉川健斗、徳留健太郎、大野行弘
  • Organizer: 第130回日本薬理学会近畿部会
  • Place of Presentation: 京都
  • Year and Date: 2016-11-19
• [Presentation] シナプス小胞蛋白SV2A遺伝子変異ラットにおける扁桃核の興奮メカニズム解析 (2016)
  • Author(s): 徳留健太郎、奥村貴裕、國澤直史、清水佐紀、金星匡人、玉田真弓、藤原汐里、三宅梨沙、芹川忠夫、真下知士、笹征史、大野行弘
  • Organizer: 第130回日本薬理学会近畿部会
  • Place of Presentation: 京都
  • Year and Date: 2016-11-19
• [Presentation] ニコチンによる運動興奮症状の発現メカニズム解析 (2016)
  • Author(s): 國澤直史、Higor A. Iha、清水佐紀、尾西美咲、野村有治、松原菜美、徳留健太郎、河合悦子、大野行弘
  • Organizer: 第66回日本薬学会近畿支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2016-10-15
• [Presentation] シナプス小胞蛋白SV2Aの遺伝子変異による扁桃核の興奮メカニズム (2016)
  • Author(s): 三宅梨沙、玉田真弓、藤原汐里、徳留健太郎、芹川忠夫、清水佐紀、大野行弘
  • Organizer: 第66回日本薬学会近畿支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2016-10-15
• [Presentation] ニコチン誘発振戦におけるセロトニン受容体の関与 (2016)
  • Author(s): 野村有治、尾西美咲、松原菜美、國澤直史、Higor A. Iha、清水佐紀、河合悦子、大野行弘
  • Organizer: 第66回日本薬学会近畿支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2016-10-15
• [Presentation] A missense mutation of the synaptic vesicle glycoprotein 2A (SV2A) gene confers seizure susceptibility by disrupting amygdala GABA release. (2016)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Naofumi Kunisawa, Saki Shimizu, Masato Kinboshi, Mayu Tamada, Shiori Fujiwara, Risa Miyake, Tadao Serikawa, Tomoji Mashimo, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第50回日本てんかん学会学術集会
  • Place of Presentation: 静岡
  • Year and Date: 2016-10-07
• [Presentation] シナプス分泌機構に着目したてんかん病態解析：シナプス小胞蛋白SV2Aとてんかん原性. (2016)
  • Author(s): 大野行弘
  • Organizer: 第50回日本てんかん学会学術集会 (シンポジウム：てんかん原性についての基礎研究：臨床に役立つ知見））
  • Place of Presentation: 静岡
  • Year and Date: 2016-10-07
  • Invited
• [Presentation] Nicotine evokes convulsive seizures by activating amygdala neurons partly through α7nACh receptors. (2016)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Takanori Nohara, Yudai Atsuta, Keito Yoshikawa, Kentaro Tokudome, Masato Kinboshi, Tadao Serikawa and Yukihiro Ohno
  • Organizer: 30th CINP World congress of Neuropsychopharmacology
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] Pharmacological analysis of nicotine-induced tremor (2016)
  • Author(s): Naofumi Kunisawa, Higor A. Iha, Saki Shimizu, Yudai Atsuta, Takanori Nohara, Kento Yoshikawa, Kentaro Tokudome, Yukihiro Ohno
  • Organizer: 30th CINP World congress of Neuropsychopharmacology
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] Enhancement of the accumbal dopamine release by the gene mutation of synaptic vesicle protein 2A (Sv2aL174Q). (2016)
  • Author(s): Kentaro Tokudome, Saki Shimizu, Naofumi Kunisawa, Azuki Hashimoto, Yuki Hiraoka, Kazuki Fukuda, Ayako Ikari, Yumiko Iguchi, Takafumi Sugahara, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 30th CINP World congress of Neuropsychopharmacology
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] Nicotine evokes convulsive seizures by activating amygdala neurons partly through α7nACh receptors (2016)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Takanori Nohara, Yudai Atsuta, Keito Yoshikawa, Kentaro Tokudome, Masato Kinboshi, Tadao Serikawa and Yukihiro Ohno
  • Organizer: The 30th CINP World congress of Neuropsychopharmacology
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] Enhancement of the accumbal dopamine release by the gene mutation of synaptic vesicle protein 2A (Sv2aL174Q) (2016)
  • Author(s): Kentaro Tokudome, Saki Shimizu, Naofumi Kunisawa, Azuki Hashimoto, Yuki Hiraoka, Kazuki Fukuda, Ayako Ikari, Yumiko Iguchi, Takafumi Sugahara, Tadao Serikawa, Yukihiro Ohno
  • Organizer: The 30th CINP World congress of Neuropsychopharmacology
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] Analysis of causative brain regions for nicotine-induced seizures. (2016)
  • Author(s): Naofumi Kunisawa, Higor A. Iha, Saki Shimizu, Yuto Mizuguchi, Takanori Nohara, Yudai Atsuta, Keito Yoshikawa, Kentaro Tokudome, Masato Kinboshi, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 第46回日本神経精神薬理学会年会
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-02
• [Presentation] Pharmacological responses of nicotine-induced tremor to various anti-tremor agents. (2016)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yudai Atsuta, Takanori Nohara, Kento Yoshikawa, Kentaro Tokudome, Yukihiro Ohno
  • Organizer: 第46回日本神経精神薬理学会年会
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-02
• [Presentation] Facilitation of kindling development by the gene mutation of synaptic vesicle protein 2A. (2016)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Tomoji Mashimo, Naofumi Kunisawa, Saki Shimizu, Ryo Terada, Masato Kinboshi, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第46回日本神経精神薬理学会年会
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-02
• [Presentation] Analysis of causative brain regions for nicotine-induced seizures (2016)
  • Author(s): Naofumi Kunisawa, Higor A. Iha, Saki Shimizu, Yuto Mizuguchi, Takanori Nohara, Yudai Atsuta, Keito Yoshikawa, Kentaro Tokudome, Masato Kinboshi, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 第46回日本神経精神薬理学会年会
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-02
• [Presentation] Facilitation of kindling development by the gene mutation of synaptic vesicle protein 2A (2016)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Tomoji Mashimo, Naofumi Kunisawa, Saki Shimizu, Ryo Terada1, Masato Kinboshi, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第46回日本神経精神薬理学会年会
  • Place of Presentation: 韓国（ソウル）
  • Year and Date: 2016-07-02
• [Presentation] Nicotine elicits limbic motor seizures primarily by activating amygdala neurons (2016)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yudai Atsuta, Takanori Nohara, Keito Yoshikawa, Kentaro Tokudome and Yukihiro Ohno
  • Organizer: 第89回日本薬理学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-09
• [Presentation] Mutation of the gene encoding synaptic vesicle protein 2A (SV2A) markedly facilitates the kindling epileptogenesis in rats (2015)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Tomoji Mashimo, Naofumi Kunisawa, Saki Shimizu, Ryo Terada, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: The 4th Congress of Asian College of Neuropsychopharmacology
  • Place of Presentation: 台湾（台北）
  • Year and Date: 2015-11-18
  • Int'l Joint Research
• [Presentation] Nicotine provokes seizures by activating amygdala neurons partly through α7-nACh receptors (2015)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Kentaro Tokudome, Masato Kinboshi, Yukihiro Ohno
  • Organizer: 第49回日本てんかん学会
  • Place of Presentation: 長崎
  • Year and Date: 2015-10-30
• [Presentation] Mechanisms underlying the kindling facilitation by the gene mutation of synaptic vesicle protein 2A (SV2A) (2015)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Naofumi Kunisawa, Kinboshi Masato, Saki Shimizu, Tomoji Mashimo, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第49回日本てんかん学会
  • Place of Presentation: 長崎
  • Year and Date: 2015-10-30
• [Presentation] Mutation of the gene encoding synaptic vesicle protein 2A (SV2A) markedly facilitates the kindling epileptogenesis in rats (2015)
  • Author(s): Naofumi Kunisawa, Kentaro Tokudome, Takahiro Okumura, Saki Shimizu, Masato Kinboshi, Tomoji Mashimo, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第49回日本てんかん学会
  • Place of Presentation: 長崎
  • Year and Date: 2015-10-30
• [Presentation] Amygdala is the primary causative site for nicotine-induced convulsive seizures (2015)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Kentaro Tokudome, Yukihiro Ohno
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28 – 2015-07-31
• [Presentation] Amygdala is the primary causative site for nicotine-induced convulsive seizures (2015)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Kentaro Tokudome, Yukihiro Ohno
  • Organizer: 第38回日本神経科学会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Presentation] ニコチン誘発けいれんの原因部位およびメカニズムの解析 (2015)
  • Author(s): 國澤直史、清水佐紀、Higor A. Iha、大高美幸、近持壽郎、高久保佑一、熱田侑大、野原孝紀、吉川健斗、徳留健太郎、河合悦子、大野行弘
  • Organizer: 第42回日本毒性学会学術年会
  • Place of Presentation: 金沢
  • Year and Date: 2015-06-29
• [Presentation] Pharmacological mechanisms of nicotine-induced convulsive seizures in mice (2015)
  • Author(s): Higor A. Iha, Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Kentaro Tokudome, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 第88回日本薬理学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-03-18 – 2015-03-20
• [Presentation] ニコチンによるけいれん発現の薬理学的解析 (2014)
  • Author(s): 國澤直史、清水佐紀、水口裕登、奥村貴裕、大高美幸、近持壽郎、高久保佑一、徳留健太郎、芹川忠夫、大野行弘
  • Organizer: 第44回日本神経精神薬理学会・第24回日本臨床精神神経薬理学会 合同学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-11-20 – 2014-11-22
• [Presentation] Evaluation of seizure susceptibility in the synaptic vesicle protein 2A (SV2A)-mutant rats (2014)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Tomoji Mashimo, Tadao Serikawa, Naofumi Kunisawa, Saki Shimizu, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第48回日本てんかん学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2014-10-02 – 2014-10-03
• [Presentation] Pharmacological analysis for the mechanism of nicotine-induced seizures in rats (2014)
  • Author(s): Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Takahiro Okumura, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Kentaro Tokudome, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 第48回日本てんかん学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2014-10-02 – 2014-10-03
• [Presentation] Analysis of brain regions responsible for nicotine-induced convulsions (2014)
  • Author(s): Naofumi Kunisawa, Saki Shimizu, Yuto Mizuguchi, Takahiro Okumura, Kentaro Tokudome, Miyuki Ohtaka, Hisao Chikamochi, Yuichi Takakubo, Tadao Serikawa, Yukihiro Ohno
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] Enhanced kindling development in synaptic vesicle protein 2A-mutant (Sv2aL174Q) rats (2014)
  • Author(s): Kentaro Tokudome, Takahiro Okumura, Tomoji Mashimo, Naofumi Kunisawa, Saki Shimizu, Ryo Terada, Tadao Serikawa, Masashi Sasa, Yukihiro Ohno
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] ニコチンによるけいれん発現のメカニズム解析 (2014)
  • Author(s): 大野行弘、國澤直史、水口裕登、奥村貴裕、徳留健太郎、落合 緑、溝辺雄輔、田村深雪、大高美幸、近持壽郎、高久保佑一、河合悦子、清水佐紀
  • Organizer: 第41回日本毒性学会学術年会
  • Place of Presentation: 神戸
  • Year and Date: 2014-07-02 – 2014-07-04
• [Book] In Vivo Neuropharmacology and Neurophysiology (2016)
  • Author(s): H.A. Iha, N. Kunisawa, K. Tokudome, T. Mukai, M. Kinboshi, S. Shimizu, Y. Ohno
  • Publisher: Springer Science+Business Media, LLC, New York
• [Remarks] 大阪薬科大学 薬品作用解析学研究室
  • URL: http://www.oups.ac.jp/kenkyu/kenkyuushitu/yakuhinsayou.html