Elucidating synaptic mechanisms and treatment of epileptic encephalopathy associated with CDKL5 mutations

• Project/Area Number: 15K09614
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: The University of Tokyo
• Principal Investigator: TANAKA Teruyuki 東京大学, 大学院医学系研究科(医学部), 准教授 (10246647)
• Co-Investigator(Renkei-kenkyūsha): MIZUGUCHI Masashi 東京大学, 大学院医学系研究科, 教授 (20209753) ; MANABE Toshiya 東京大学, 医科学研究所, 教授 (70251212) ; FUKAYA Masahiro 北里大学, 医学部, 講師 (10360900) ; TAKAO Keizo 富山大学, 研究推進機構研究推進総合支援センター, 講師 (80420397)
• Research Collaborator: OKUDA Kosuke 東京大学, 大学院医学系研究科, 大学院学生 ; MURAKAMI Takuto 東京大学, 大学院医学系研究科, 大学院学生
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: CDKL5 / ノックアウトマウス / 海馬 / NMDA受容体 / GluN2B / てんかん / けいれん / キナーゼ / シナプス / 発達障害 / レット症候群

Outline of Final Research Achievements

Mutations in the CDKL5 gene cause severe epileptic encephalopathy. Cdkl5 KO mice showed normal sensitivity to kainic acid; however, they displayed significant hyperexcitability to NMDA. Electrophysiological analysis in the hippocampus disclosed an increased ratio of NMDA/AMPA receptor-mediated excitatory postsynaptic currents (EPSCs) and a significantly larger decay time constant of NMDA receptor-mediated EPSCs in Cdkl5 KO mice. Subcellular fractionation of the hippocampus from Cdkl5 KO mice revealed a significant increase of GluN2B and SAP102 in the postsynaptic density fraction. Immunoelectron microscopic analysis of the hippocampus further confirmed postsynaptic overaccumulation of GluN2B and SAP102 in Cdkl5 KO mice. Ifenprodil abrogated the NMDA-induced hyperexcitability in Cdkl5 KO mice. These data indicate that CDKL5 plays an important role in controlling postsynaptic localization of the GluN2B-SAP102 complex in the hippocampus and thereby regulates seizure susceptibility.

Research Products

• [Journal Article] Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory (2018)
  • Author(s): Okuda Kosuke、Takao Keizo、Watanabe Aya、Miyakawa Tsuyoshi、Mizuguchi Masashi、Tanaka Teruyuki
  • Journal Title: PLOS ONE Volume: 13 Issue: 4 Pages: e0196587-e0196587
  • DOI: 10.1371/journal.pone.0196587
  • Peer Reviewed / Open Access
• [Journal Article] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility. (2017)
  • Author(s): Okuda K, Kobayashi S, Fukaya M, Watanabe A, Murakami T, Hagiwara M, Sato T, Ueno H, Ogonuki N, Komano-Inoue S, Manabe H, Yamaguchi M, Ogura A, Asahara H, Sakagami H, Mizuguchi M, Manabe T, Tanaka T
  • Journal Title: Neurobiology of Diseases Volume: 106 Pages: 158-170
  • DOI: 10.1016/j.nbd.2017.07.002
  • Peer Reviewed / Open Access
• [Journal Article] Towards elucidating the regulatory roles of <i>CDKL5</i> in synaptic transmission (2015)
  • Author(s): 奥田耕助、田中輝幸
  • Journal Title: Folia Pharmacologica Japonica Volume: 145 Issue: 4 Pages: 183-186
  • DOI: 10.1254/fpj.145.183
  • NAID: 130005061553
  • ISSN: 0015-5691, 1347-8397
• [Presentation] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus, and regulates seizure susceptibility, emotional behaviors, and memory (2018)
  • Author(s): 田中 輝幸, 奥田 耕助, 小林 静香, 村上 拓冬, 深谷 昌弘, 高雄 啓三, 渡邉 紀, 萩原 舞, 阪上 洋行, 水口 雅, 宮川 剛, 真鍋 俊也
  • Organizer: The 8th BRI International Symposium 2018, Niigata, Japan
  • Int'l Joint Research
• [Presentation] Comprehensive analyses of CDKL5, a causative gene product for neurodevelopment disorders West syndrome or atypical Rett syndrome (2017)
  • Author(s): 田中 輝幸, 奥田 耕助, 小林 静香, 村上 拓冬, 深谷 昌弘, 高雄 啓三, 渡邉 紀, 萩原 舞, 阪上 洋行, 水口 雅, 宮川 剛, 真鍋 俊也
  • Organizer: 第40回日本神経科学大会、千葉市
• [Presentation] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility (2017)
  • Author(s): T. TANAKA, K. OKUDA, S. KOBAYASHI, M. FUKAYA, K. TAKAO, A. WATANABE, T. MURAKAMI, M. HAGIWARA, S. KOMANO-INOUE, H. MANABE, M. YAMAGUCHI, H. SAKAGAMI, T. MIYAKAWA, M. MIZUGUCHI, T. MANABE
  • Organizer: 2017 Annual Meeting of the Society for Neuroscience, Washington, DC, USA
  • Int'l Joint Research
• [Presentation] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility (2017)
  • Author(s): T. TANAKA, K. OKUDA, S. KOBAYASHI, M. FUKAYA, K. TAKAO, A. WATANABE, T. MURAKAMI, M. HAGIWARA, S. KOMANO-INOUE, H. MANABE, M. YAMAGUCHI, H. SAKAGAMI, T. MIYAKAWA, M. MIZUGUCHI, T. MANABE
  • Organizer: 2017 CDKL5 Forum Meeting, Boston, USA
  • Int'l Joint Research
• [Presentation] CDKL5 controls postsynaptic NMDA receptor localization and regulates memory, emotion and seizure susceptibility (2016)
  • Author(s): 村上拓冬、奥田耕助、小林静香、深谷昌弘、高雄啓三、渡邉紀、萩原舞、阪上洋行、水口雅、宮川剛、真鍋俊也、田中輝幸
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜会議センター（神奈川県横浜市）
• [Presentation] CDKL5 controls postsynaptic NMDA receptor localization and regulates memory, emotion and seizure susceptibility (2016)
  • Author(s): 村上拓冬、奥田耕助、小林静香、深谷昌弘、高雄啓三、近江信次、渡邉紀、萩原舞、阪上洋行、水口雅、宮川剛、真鍋俊也、田中輝幸
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜会議センター（神奈川県横浜市）
• [Presentation] West症候群・Rett症候群の原因遺伝子CDKL5は、NMDA受容体機能を制御し、記憶・学習、情動、易痙攣性を調節する (2015)
  • Author(s): 田中輝幸、奥田耕助、小林静香、村上拓冬、深谷昌弘、高雄啓三、渡邉紀、萩原舞、阪上洋行、水口雅、宮川剛、真鍋俊也
  • Organizer: 第4５回日本神経精神薬理学会・第37回日本生物学的精神医学会
  • Place of Presentation: タワーホール船堀 (東京都江戸川区)
  • Year and Date: 2015-09-25
• [Presentation] CDKL5 controls NMDA receptor function and regulates memory, emotion and seizure susceptibility (2015)
  • Author(s): 田中 輝幸、奥田 耕助、小林 静香、深谷 昌弘、高雄 啓三、 渡邉 紀、村上 拓冬、萩原 舞、阪上 洋行、水口 雅、宮川 剛、真鍋 俊也
  • Organizer: 第38回日本神経科学大会 Neuro 2015
  • Place of Presentation: 神戸国際会議場 (兵庫県神戸市)
  • Year and Date: 2015-07-28
• [Presentation] West 症候群/Rett 症候群の原因遺伝子 CDKL5 は、 グルタミン酸受容体局在を制御し 記憶/情動/易痙攣性を調節する (2015)
  • Author(s): 田中 輝幸、奥田 耕助、小林 静香、深谷 昌弘、高雄 啓三、 渡邉 紀、村上 拓冬、萩原 舞、阪上 洋行、水口 雅、宮川 剛、真鍋 俊也
  • Organizer: 第55回日本先天異常学会学術集会・第38回日本小児遺伝学会学術集会
  • Place of Presentation: パシフィコ横浜会議センター (神奈川県横浜市)
  • Year and Date: 2015-07-25
• [Presentation] CDKL5 controls glutamate receptor function and regulates memory, emotion and seizure susceptibility (2015)
  • Author(s): 田中輝幸、水口雅
  • Organizer: 第57回日本小児神経学会総会
  • Place of Presentation: 帝国ホテル 大阪 (大阪府大阪市)
  • Year and Date: 2015-05-28