Changes in GABAergic inputs to GnRH neurons causing stress induced reproductive dysfunction

• Project/Area Number: 16K08523
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Environmental physiology(including physical medicine and nutritional physiology)
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: Watanabe Miho 浜松医科大学, 医学部, 助教 (10399321)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: GABA / GnRHニューロン / KCC2 / 生殖 / 視床下部 / クロライド / NKCC1 / 神経内分泌 / ストレス / 神経内分泌学

Outline of Final Research Achievements

The GnRH neurosecretory system constitutes the final common pathway in the neuroendocrine control of reproduction. GABA acts excitatory on GnRH neurons. Overexpression of KCC2 inhibited calcium oscillations in GnRH neurons. Stress may cause the disruption of intracellular chloride homeostasis in GnRH neurons, thereby lead to reproductive dysfunction.

Academic Significance and Societal Importance of the Research Achievements

本研究ではGnRHニューロンの機能不全による生殖機能低下のメカニズムの一端を明らかにすることができた。ストレスや栄養状態の低下による体重減少などにより、視床下部-下垂体の機能が低下し、無月経や無排卵になる中枢性排卵障害の原因の一つにGnRHニューロンの機能不全が考えられる。ヒトのGnRHニューロンへもGABA入力があることがわかっており、細胞内クロライド濃度維持機構の破綻とGnRHニューロンの機能不全との関連性を示唆した本研究の成果は無排卵や不妊症などの治療、新たな治療薬開発への貢献が考えられる。

Research Products

• [Int'l Joint Research] Hungarian Academy of Sciences(ハンガリー)
• [Int'l Joint Research] Yale School of Medicine(米国)
• [Int'l Joint Research] Hungarian Academy of Sciences(ハンガリー)
• [Journal Article] Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival. (2019)
  • Author(s): Watanabe M, Zhang J, Mansuri MS, Duan J, Karimy JK, Delpire E, Alper SL, Lifton RP, Fukuda A, Kahle KT.
  • Journal Title: Science Signaling Volume: 12 Issue: 603
  • DOI: 10.1126/scisignal.aaw9315
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Pathogenic potential of human SLC12A5 variants causing KCC2 dysfunction. (2019)
  • Author(s): Fukuda A and Watanabe M
  • Journal Title: Brain Research Volume: 1710 Pages: 1-7
  • DOI: 10.1016/j.brainres.2018.12.025
  • Peer Reviewed / Open Access
• [Journal Article] Time-controllable Nkcc1 knockdown replicates reversible hearing loss in postnatal mice. (2017)
  • Author(s): Watabe T, Xu M, Watanabe M, Nabekura J, Higuchi T, Hori K, Sato MP, Nin F, Hibino H, Ogawa K, Masuda M, Tanaka KF
  • Journal Title: Science Reports Volume: 7 Issue: 1 Pages: 13605-14007
  • DOI: 10.1038/s41598-017-13997-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A novel GABA-mediated corticotropin-releasing hormone secretory mechanism in the median eminence. (2016)
  • Author(s): Kakizawa K, Watanabe M, Mutoh H, Okawa Y, Yamashita M, Yanagawa Y, Itoi K, Suda T, Oki Y. and Fukuda A.
  • Journal Title: Science Advances Volume: 2 Issue: 8 Pages: 1-15
  • DOI: 10.1126/sciadv.1501723
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Impaired neuronal KCC2 function by biallelic SLC12A5 mutations in migrating focal seizures and severe developmental delay. (2016)
  • Author(s): Saitsu H, Watanabe M, Akita T, Ohba C, Sugai K, Ong WP, Shiraishi H, Yuasa S, Matsumoto H, Beng KT, Saitoh S, Miyatake S, Nakashima M, Miyake N, Kato M, Fukuda A. and Matsumoto N.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 30072-30072
  • DOI: 10.1038/srep30072
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival. (2020)
  • Author(s): Watanabe M, Kahle KT, Fukuda A
  • Organizer: 第97回日本生理学会大会
• [Presentation] 発達期のCRHニューロン制御機構におけるGABAの役割 (2020)
  • Author(s): 渡部美穂、柿沢圭亮、福田敦夫
  • Organizer: NINS分野融合型共同事業ワークショップ
• [Presentation] Excitatory GABAergic inputs to GnRH neurons are required for estrous cyclicity. (2019)
  • Author(s): Watanabe M, Fukuda A
  • Organizer: The 19th Kyungpook-Hamamatsu Joint Medical Symposium Hamamatsu Meeting
  • Int'l Joint Research
• [Presentation] Excitatory GABAergic inputs to GnRH neurons are required for female reproduction. (2019)
  • Author(s): Watanabe M, Fukuda A
  • Organizer: Society for Neuroscience 49th Annual Meeting
  • Int'l Joint Research
• [Presentation] 抑制性神経伝達を維持するカリウム-クロライド共役担体(KCC2)のリン酸化による制御の役割 (2019)
  • Author(s): 渡部美穂、Kahle KT、福田敦夫
  • Organizer: 第66回中部生理学会
• [Presentation] A KCC2 phospho-switch is essential for dynamic GABA-mediated inhibition and postnatal survival. (2019)
  • Author(s): Watanabe M, Zhang J, Mansuri M, Duan J, Kahle KT, Fukuda A
  • Organizer: 第42回日本神経科学大会、第62回日本神経化学会大会、新潟
• [Presentation] Excitatory GABAergic inputs to GnRH neurons are required for female reproduction. (2019)
  • Author(s): Watanabe M, Nabekura J, Fukuda A
  • Organizer: The 18th Kyungpook-Hamamatsu Joint Medical Symposium Hamamatsu Meeting
  • Int'l Joint Research
• [Presentation] Developmental regulation of KCC2 phosphorylation is essential for GABA signaling and survival. (2019)
  • Author(s): Watanabe M, Zhang J, Mansuri M, Duan J, Kahle KT, Fukuda A
  • Organizer: 9th Federation of the Asian and Oceanian Physiological Societies Congress
  • Int'l Joint Research
• [Presentation] CRH release regulation by GABAergic projection from arcuate nucleus using chemogenetic model. (2019)
  • Author(s): Yesmin R, Watanabe M, Fukuda A
  • Organizer: 9th Federation of the Asian and Oceanian Physiological Societies Congress
  • Int'l Joint Research
• [Presentation] GABAによる抑制性伝達におけるカリウム-クロライド共役担体(KCC2)のリン酸化による機能制御の役割 (2018)
  • Author(s): 渡部美穂、Zhang J、Mansuri M、Duan J、Kahle KT、福田敦夫
  • Organizer: 第65回中部生理学会
• [Presentation] Development of chemogenetic model to study physiological roles of the GABAergic projection from arcuate nucleus to the CRH nerve terminals (2018)
  • Author(s): Yesmin R, Watanabe M, Fukuda A
  • Organizer: 第65回中部生理学会
• [Presentation] 生殖腺刺激ホルモン放出ホルモン(GnRH)ニューロン特異的にKCC2を発現させたマウスでは生殖機能に異常がみられる (2018)
  • Author(s): 渡部美穂、鍋倉淳一、福田敦夫
  • Organizer: 第95回日本生理学会
• [Presentation] The role of excitatory action of GABA in the regulation of reproduction (2017)
  • Author(s): Watanabe M, Nabekura J, Fukuda A
  • Organizer: The 94th Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Hamamatsu
  • Year and Date: 2017-03-30
• [Presentation] Mild functional impairment of neuronal K+-Cl－ cotransporter KCC2 by biallelic mutations causes migrating focal seizures and severe developmental delay (2017)
  • Author(s): Akita T, Saitsu H, Watanabe M, Matsumoto N, Fukuda A
  • Organizer: The 94th Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Hamamatsu
  • Year and Date: 2017-03-29
• [Presentation] Constitutive phosphomimetic inhibition of KCC2 at Thr906/Thr1007 causes GABAdependent network excitability, seizure, and early postnatal death. (2017)
  • Author(s): Watanabe M, Duan J, Mansuri M. Zhang J, Fukuda A, Kahle KT.
  • Organizer: Society for Neuroscience 47th Annual Meeting
  • Int'l Joint Research
• [Presentation] KCC2 dysfunction underlying infantile and neonatal seizures in an animal model. (2017)
  • Author(s): Fukuda A, Watanabe M, Akita T.
  • Organizer: 第51回日本てんかん学会学術集会
• [Presentation] 生殖腺刺激ホルモン放出ホルモン(GnRH)ニューロンの制御における興奮性GABA入力の役割 (2017)
  • Author(s): 渡部美穂、鍋倉淳一、福田敦夫
  • Organizer: 第64回中部日本生理学会
• [Presentation] Conditional upregulation of the KCC2 membrane transporter in a transgenic mouse and the effects on neuronal excitability in vitro (2016)
  • Author(s): Goulton CS, Wang K, Khoshaba A, Watanabe M, Nabekura J, Moorhouse AJ
  • Organizer: Society for Neuroscience 46h Annual Meeting
  • Place of Presentation: San Diego
  • Year and Date: 2016-12-16
• [Presentation] The effect of upregulated KCC2 expression on chemically induced seizures and diazepam therapy in vivo (2016)
  • Author(s): Cheung DL, Goulton CS, Watanabe M, Nabekura J, Moorhouse AJ
  • Organizer: Society for Neuroscience 46h Annual Meeting
  • Place of Presentation: San Diego
  • Year and Date: 2016-12-16
• [Presentation] 乳児焦点移動性部分発作はSLC12A5遺伝子の両アレル変異によるカリウム-クロライド共役担体(KCC2)機能の低下により引き起こされる (2016)
  • Author(s): 渡部美穂、秋田天平、才津浩智、松本直通、福田敦夫
  • Organizer: 第63回中部生理学会
  • Place of Presentation: Okazaki
  • Year and Date: 2016-11-04
• [Presentation] Impaired neuronal KCC2 function by biallelic SLC12A5 mutations in migrating focal seizures and severe developmental delay (2016)
  • Author(s): Watanabe M, Akita T, Saitsu H, Matsumoto N, Fukuda A
  • Organizer: The 13th Korea-Japan Joint Symposium of Brain Sciences, and Cardiac and Smooth Muscle Sciences
  • Place of Presentation: Saga
  • Year and Date: 2016-08-24
• [Presentation] A novel excitatory GABAergic input from the arcuate nucleus to the median eminence involved in CRH release (2016)
  • Author(s): Fukuda A, Kakizawa K, Watanabe M, Mutoh H, Okawa Y, Yamashita M, Yanagawa Y, Itoi K, Suda T, Oki Y
  • Organizer: The 13th Korea-Japan Joint Symposium of Brain Sciences, and Cardiac and Smooth Muscle Sciences
  • Place of Presentation: Saga
  • Year and Date: 2016-08-22
• [Presentation] Partial loss of neuronal KCC2 function by biallelic SLC12A5 mutations causes migrating focal seizures and developmental delay (2016)
  • Author(s): 渡部美穂、秋田天平、才津浩智、松本直通、福田敦夫
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: Yokohama
  • Year and Date: 2016-07-22
• [Presentation] The excitatory GABA action at the median eminence maintains the steady-state release of corticotropin-releasing hormone (2016)
  • Author(s): Fukuda A, Kakizawa K, Watanabe M, Mutoh H, Okawa M, Yamashita M, Yanagawa Y, Itoi K, Suda T, Oki Y
  • Organizer: 10th Forum of European Neuroscience
  • Place of Presentation: Copenhagen
  • Year and Date: 2016-07-03
• [Book] Neuronal Chloride Transporters in Health and Disease Chapter 11. Post-translational modification of neuronal chloride transporters (2020)
  • Author(s): Miho Watanabe, Atsuo Fukuda
  • Publisher: Elsevier
• [Remarks] 神経生理学講座
  • URL: https://www.hama-med.ac.jp/education/fac-med/dept/neurophysiology/index.html
• [Remarks] 浜松医科大学神経生理学講座 研究活動
  • URL: https://www.hama-med.ac.jp/education/fac-med/dept/neurophysiology/res-act/index.html
• [Remarks] 神経生理学講座ホームページ
  • URL: https://www.hama-med.ac.jp/uni_education_igakubu_igaku_seiri1.html