Developmental disorder model based on fetal hypothalamic GABA-Cl system disturbances

• Project/Area Number: 17K19682
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Internal medicine of the bio-information integration and related fields
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: FUKUDA Atsuo 浜松医科大学, 医学部, 教授 (50254272)
• Research Collaborator: AKITA Tenpei ; WATANABE Miho ; HATA Kenichiro
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 発達障害 / 低栄養 / 胎児脳 / ストレス / 視床下部 / 精神疾患 / KCC2 / CRH

Outline of Final Research Achievements

We confirmed that c-fos expression was elevated in AgRP cells in the arcuate nucleus and in CRH cells in the paraventricular nucleus by feeding restriction, thus established evaluation methods for both known and novel HPA axis reaction in maternal malnourished fetuses. In the maternal malnutrition model for genetically modified mice, food restriction (-30%) was performed on pregnancy at 10.5-18.5 days, establishing a fetal malnutrition mouse model, and the developmental state of AgRP cells and CRH cells were evaluated in the fetal brains. In addition, it was suggested that proper suppression of the phosphorylation of Thr906 and Thr1007 of KCC2 during the fetal stage is important for the normal neurogenesis and migration of neurons. Prenataly-stressed GAD67 heterozygous fetuses were found to show some of psychiatric disease phenotypes.

Academic Significance and Societal Importance of the Research Achievements

精神的ストレスや低栄養ではHPA軸の活動が高まり、室傍核のCRH細胞に投射する抑制性GABA神経は細胞内Cl-濃度依存的にHPA軸活動を変化させる。胎児期の母親の低栄養や子のGABA神経系の異常は精神神経疾患のリスクとして知られるが、胎児のHPA軸機能がGABA作用変化により受ける影響はわかっていない。そこで、母体に食餌制限を行ったり、CRH細胞やAgRP細胞（GABA陽性）で遺伝子操作をして、胎仔のHPA軸反応、神経細胞の発生・移動及び生後の発現型を評価した。母体低栄養やストレスが胎仔GABAシステムを介して、HPA軸に作用して精神神経疾患様の発現型を誘導することを明らかにした。

Research Products

• [Int'l Joint Research] Yale School of Medicine(米国)
• [Int'l Joint Research] マインツ大学(ドイツ)
• [Int'l Joint Research] Yale School of Medicine(米国)
• [Int'l Joint Research] マインツ大学(ドイツ)
• [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] Alterations of GABAergic Neuron-Associated Extracellular Matrix and Synaptic Responses in Gad1-Heterozygous Mice Subjected to Prenatal Stress (2018)
  • Author(s): Wang T, Sinha AS, Akita T, Yanagawa Y, Fukuda A
  • Journal Title: Frontiers in Cellular Neuroscience Volume: 12 Pages: 284-284
  • DOI: 10.3389/fncel.2018.00284
  • Peer Reviewed / Open Access
• [Journal Article] MHC class I in dopaminergic neurons suppresses relapse to reward seeking (2018)
  • Author(s): Murakami G, Edamura M, Furukawa T, Kawasaki H, Kosugi I, Fukuda A, Iwashita T, Nakahara D
  • Journal Title: Science Advances Volume: 4 Issue: 3 Pages: 7388-7388
  • DOI: 10.1126/sciadv.aap7388
  • Peer Reviewed / Open Access
• [Journal Article] Optical measurement of glutamate in slice preparations of the mouse retina (2018)
  • Author(s): M. Ohkuma, M. Kaneda, S. Yoshida, A. Fukuda, E. Miyachi
  • Journal Title: Neuroscience Research Volume: 印刷中 Pages: 30544-8
  • DOI: 10.1016/j.neures.2018.03.001
  • Peer Reviewed / Open Access
• [Journal Article] Biallelic Variants in CNPY3 , Encoding an Endoplasmic Reticulum Chaperone, Cause Early-Onset Epileptic Encephalopathy (2018)
  • Author(s): Mutoh Hiroki、Kato Mitsuhiro、Akita Tenpei、Shibata Takuma、Wakamoto Hiroyuki、Ikeda Hiroko、Kitaura Hiroki、Aoto Kazushi、Nakashima Mitsuko、Wang Tianying、Ohba Chihiro、Miyatake Satoko、Miyake Noriko、Kakita Akiyoshi、Miyake Kensuke、Fukuda Atsuo、Matsumoto Naomichi、Saitsu Hirotomo
  • Journal Title: The American Journal of Human Genetics Volume: 102 Issue: 2 Pages: 321-329
  • DOI: 10.1016/j.ajhg.2018.01.004
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] De novo variants in CAMK2A and CAMK2B cause neurodevelopmental disorders (2018)
  • Author(s): Akita Tenpei、Aoto Kazushi、Kato Mitsuhiro、Shiina Masaaki、Mutoh Hiroki、Nakashima Mitsuko、Kuki Ichiro、Okazaki Shin、Magara Shinichi、Shiihara Takashi、Yokochi Kenji、Aiba Kaori、Tohyama Jun、Ohba Chihiro、Miyatake Satoko、Miyake Noriko、Ogata Kazuhiro、Fukuda Atsuo、Matsumoto Naomichi、Saitsu Hirotomo
  • Journal Title: Annals of Clinical and Translational Neurology Volume: 5 Issue: 3 Pages: 280-296
  • DOI: 10.1002/acn3.528
  • Peer Reviewed / Open Access
• [Journal Article] Taurine as an Essential Neuromodulator during Perinatal Cortical Development (2017)
  • Author(s): Kilb W, Fukuda A.
  • Journal Title: Frontiers in Cellular Neuroscience Volume: 11:328 Pages: 328-328
  • DOI: 10.3389/fncel.2017.00328
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Human epilepsy and animal model with mutations in KCC2. (2019)
  • Author(s): Fukuda A.
  • Organizer: 9th FAOPS
  • Int'l Joint Research
• [Presentation] Ca2+ signaling and ion channel activation in embryonic neurons in the medial ganglionic eminence. (2019)
  • Author(s): Akita T, Fukuda A.
  • Organizer: 9th FAOPS
  • 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 FAOPS
  • Int'l Joint Research
• [Presentation] Effect of orexin on the firing pattern of serotonergic dorsal raphe neurons. (2019)
  • Author(s): Ishibashi M, Molina NE, Fukuda A.
  • Organizer: 9th FAOPS
  • Int'l Joint Research
• [Presentation] Prenatal stress on Gad1-heterozygotes perturbs development of GABAergic networks affecting behavior. (2019)
  • Author(s): Wang T, Sinha AS, Mutoh H, Akita T, Yanagawa Y, Kawai T, Hata K, Fukuda A.
  • Organizer: 9th FAOPS
  • Int'l Joint Research
• [Presentation] Mechanisms underlying WNK3 kinase mediated regulation of neuronal excitability in prefrontal cortex. (2019)
  • Author(s): Sinha AS, Wang T, Hosoi Y, Sohara E, Akita T, Uchida S, Fukuda A.
  • Organizer: 9th FAOPS
  • 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 FAOPS
  • Int'l Joint Research
• [Presentation] Gad1-heterozygotes subjected to prenatal stress undergo alterations in extracellular matrix associated with GABAergic neuron and inhibitory synaptic transmission. (2019)
  • Author(s): Sinha AS, Wang T, Akita T, Yanagawa Y, Fukuda A.
  • Organizer: 第25回浜松医科学シンポジウム
• [Presentation] 細胞内タウリンによるPKC基質部位リン酸化を介したGABAA受容体膜移行維持機構. (2019)
  • Author(s): 細井泰志, 渡部美穂, 秋田天平, 伊藤崇志, 福田敦夫.
  • Organizer: 第5回国際タウリン研究会日本部会
• [Presentation] 胎生期大脳基底核原基の神経前駆細胞内Ca2+シグナリングについて. (2018)
  • Author(s): 秋田天平, 福田敦夫.
  • Organizer: 第65回中部日本生理学会
• [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回中部日本生理学会
• [Presentation] 背側縫線核セロトニン作動性ニューロンにおけるオレキシン誘発性の発火後過分極増強と内向き電流への細胞外カルシウムの影響. (2018)
  • Author(s): 石橋賢, Molina NE, 福田敦夫, Leonard CS.
  • Organizer: 第65回中部日本生理学会
• [Presentation] Alterations of GABAergic Neuron-Associated Extracellular Matrix and Synaptic Responses in Gad1-Heterozygous Mice Subjected to Prenatal Stress. (2018)
  • Author(s): Sinha AS, Wang T, Mutoh H, Akita T, Yanagawa Y, Fukuda A.
  • Organizer: 第65回中部日本生理学会
• [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回中部日本生理学会
• [Presentation] Effect of orexin on the firing pattern of serotonergic dorsal raphe neurons. (2018)
  • Author(s): Ishibashi M, Molina NE, Fukuda A, Leonard CS.
  • Organizer: 第45回日本脳科学会
• [Presentation] てんかんにおけるGABAシナプスの役割. (2018)
  • Author(s): 福田敦夫.
  • Organizer: 第61回日本脳循環代謝学会学術集会
  • Invited
• [Presentation] Prenatal stress to the GAD67 heterozygous mice impairs GABA neurogenesis followed by physiological dysfunctions in postnatal life. (2018)
  • Author(s): Fukuda A.
  • Organizer: 第40回日本生物学的精神医学会・第61回日本神経化学会大会 合同年会
  • Invited
• [Presentation] Prenatal stress on Gad1-heterozygotes selectively perturbs GABAergic neurogenesis, GABAergic synapse function and behavioral phenotypes. (2018)
  • Author(s): Wang T, Sinha AS, Mutoh H, Akita T, Yanagawa Y, Kawai T, Hata K, Fukuda A.
  • Organizer: 第7回日本DOHaD学会
• [Presentation] Cl-ホメオダイナミクスとマルチモーダルGABA:脳での多彩な働き. (2018)
  • Author(s): 福田敦夫.
  • Organizer: 第65回脳の医学・生物学研究会
  • Invited
• [Presentation] WNK3 Kinase regulates inwardly rectifying potassium channels in layer V pyramidal neurons of the mouse prefrontal cortex. (2018)
  • Author(s): Sinha AS, Hosoi Y, Sohara E, Mutoh H, Akita T, Uchida S, Fukuda A.
  • Organizer: 第41回日本神経科学大会
• [Presentation] Prenatal stress on Gad1-heterozygotes selectively perturbs GABAergic neurogenesis, GABAergic synapse function and behavioral phenotypes. (2018)
  • Author(s): Wang T, Sinha AS, Mutoh H, Akita T, Yanagawa Y, Kawai T, Hata K, Fukuda A.
  • Organizer: 第41回日本神経科学大会
• [Presentation] De novo mutants of CaMKIIα/β responsible for neurodevelopmental disorders upregulate A-type voltage-dependent K+ currents in hippocampal neurons. (2018)
  • Author(s): Akita T, Aoto K, Kato M, Shiina M, Mutoh H, Nakashima M, Kuki I, Okazaki S, Magara S, Shiihara T, Yokochi K, Aiba K, Tohyama J, Ohba C, Miyatake S, Miyake N, Ogata K, Fukuda A, Matsumoto N, Saitsu H.
  • Organizer: 第41回日本神経科学大会
• [Presentation] Regulated Phosphorylation of KCC2 at Thr906/Thr1007 is essential for activity-dependent Cl-extrusion during normal brain development. (2018)
  • Author(s): Fukuda, A.
  • Organizer: FENS forum 2018
  • Int'l Joint Research
• [Presentation] 神経・精神疾患の病因・病態仮説としてのマルチモーダル GABA. (2018)
  • Author(s): 福田敦夫.
  • Organizer: お茶の水ニューロサイエンスセミナー
  • Invited
• [Presentation] Maternal taurine modulate fetal brain development via intracellular and extracellular effectors. (2018)
  • Author(s): Fukuda, A.
  • Organizer: The 21st International Taurine Meeting
  • Int'l Joint Research
• [Presentation] Diverse Actions of Astrocytes at GABAergic Tripartite Synapses in Seizures: Are they protective or exacerbative? (2018)
  • Author(s): Fukuda, A.
  • Organizer: 2018 Symposium on Functional Interactions among Glia and Neurons
  • Int'l Joint Research / Invited
• [Presentation] De novo CaMKⅡα/β mutants causing neurodevelopmental disorders upregulate A-type voltage-dependent K+ currents in hippocampal neurons. (2018)
  • Author(s): Akita T, Aoto K, Fukuda A, Matsumoto N, Saitsu H.
  • Organizer: 第95回日本生理学会大会
• [Presentation] WNK3 kinase regulates inwardly rectifying potassium current in layer V pyramidal neurons of the mouse prefrontal cortex. (2018)
  • Author(s): Sinha Adya S, Hosoi Y, Sohara E, Mutoh H, Akita T, Uchida S, Fukuda A.
  • Organizer: 第95回日本生理学会大会
• [Presentation] Temporally genetic expression of KCC2 in GnRH neurons in vivo causes impairment of fertility. (2018)
  • Author(s): Watanabe M, Nabekura J, Fukuda A.
  • Organizer: 第95回日本生理学会大会
• [Presentation] タウリン欠乏はGABAA受容体のリン酸化状態を変化させることにより大脳皮質感覚野錐体細胞におけるポストシナプスのGABAA受容体数を減少させる. (2018)
  • Author(s): 細井泰志, 渡部美穂, 秋田天平, 武藤弘樹, 伊藤崇志, 福田敦夫.
  • Organizer: 第4回国際タウリン研究会日本部会
• [Presentation] The role of GABAergic interneuron-astrocyte interaction in epileptic seizures. (2018)
  • Author(s): Mutoh, H., Fukuda, A.
  • Organizer: 第１回山梨大学先端脳科学特別教育プログラム国際シンポジウム
  • Int'l Joint Research
• [Presentation] GABA synaptic dynamics and astrocytic Cl- buffering underlying neural network oscillation. (2017)
  • Author(s): 福田敦夫
  • Organizer: 第47回日本臨床神経生理学会学術大会
• [Presentation] A novel CRH secretory mechanism by an excitatory GABAergic input from the arcuate nucleus to the median eminence. (2017)
  • Author(s): Fukuda, A.
  • Organizer: The 17th Hamamatsu-Kyungpook Joint Medical Symposium
  • Int'l Joint Research / Invited
• [Presentation] Regulation of inwardly rectifying potassium current by WNK3 kinase in layer V pyramidal neurons of the mouse prefrontal cortex. (2017)
  • Author(s): Sinha Adya, S., Hosoi, Y., Sohara, E., Mutoh, H., Akita, T., Uchida, S., Fukuda, A.
  • Organizer: The 17th Hamamatsu-Kyungpook Joint Medical Symposium
  • Int'l Joint Research
• [Presentation] Prenatal stress on Gad1-heterozygotes perturbs GABAergic neurogenesis, GABAergic synapse formation and behavioral phenotypes. (2017)
  • Author(s): Fukuda, A. Wang, T., Sinha Adya, S., Yanagawa, Y., Kawai, T. and Hata, K.
  • Organizer: Neuroscience 2017
  • Int'l Joint Research
• [Presentation] Constitutive phosphomimetic inhibition of KCC2 at Thr906/Thr1007 causes GABA-dependent network excitability, seizure, and early postnatal death. (2017)
  • Author(s): Watanabe, M., Duan, J., Mansuri, M., Zhang, J., Fukuda, A. and Kahle, K.T.
  • Organizer: Neuroscience 2017
  • Int'l Joint Research
• [Presentation] KCC2 dysfunction underlying infantile and neonatal seizures in an animal model. (2017)
  • Author(s): 福田敦夫, 渡部美穂, 秋田天平, Kahle, K.
  • Organizer: 第51回日本てんかん学会学術集会
• [Presentation] Prenatal stress on Gad1- heterozygotes affects inhibitory synaptic transmission. (2017)
  • Author(s): Wang, T., Sinha Adya, S., Akita, T., Yanagawa, Y., Fukuda, A.
  • Organizer: 生理学研究所研究会「シナプス・神経回路機能の時空間制御」
• [Presentation] Prenatal stress on Gad1-heterozygotes perturbs development of brain and behavior by epigenetic mechanism. (2017)
  • Author(s): Wang T, Sinha AS, Yanagawa Y, Kawai T, Hata K and Fukuda A.
  • Organizer: 第44回日本脳科学会
• [Presentation] Regulation of inwardly rectifying potassium current by WNK3 kinase in layer V pyramidal neurons of the mouse prefrontal cortex. (2017)
  • Author(s): Sinha Adya, S., Hosoi, Y., Sohara, E., Mutoh, H., Akita, T., Uchida, S. and Fukuda, A.
  • Organizer: 第64回中部日本生理学会
• [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] てんかん原性獲得過程における抑制性神経とグリア細胞の相互作用. (2017)
  • Author(s): 武藤弘樹, 福田敦夫
  • Organizer: 第64回中部日本生理学会
• [Presentation] A novel CRH secretory mechanism by GABAergic input from the arcuate nucleus to the axon terminals of high [Cl-]i. (2017)
  • Author(s): Fukuda, A.
  • Organizer: Seminar of Center for Neuron-Glia Interaction, KIST(Korea Institute of Science and Technology)
  • Int'l Joint Research / Invited
• [Presentation] Taurine depletion reduces postsynaptic GABAA receptors in layer 2/3 pyramidal neurons of the somatosensory cortex. (2017)
  • Author(s): Hosoi, Y., Akita, T., Watanabe, M., Mutoh, H., Miyajima, H. and Fukuda, A.
  • Organizer: XXIII World Congress of Neurology（WCN2017）
  • Int'l Joint Research
• [Presentation] 母体ストレスとGABAが仔脳発達に与える病態生理学的影響. (2017)
  • Author(s): 福田敦夫, 王天英, Sinha Adya S, 柳川右千夫, 河合智子, 秦健一郎.
  • Organizer: 第27回日本病態生理学会大会
• [Presentation] Regulation of inwardly rectifying potassium current by WNK3 kinase in layer V pyramidal neurons of the mouse prefrontal cortex. (2017)
  • Author(s): Sinha Adya, S., Hosoi,Y., Sohara, E., Mutoh, H., Akita, T., Uchida, S. and Fukuda, A.
  • Organizer: 第40回日本神経科学大会
• [Presentation] The role of GABAergic interneuron-astrocyte interaction in epileptic seizures. (2017)
  • Author(s): Mutoh, H., Fukuda, A.
  • Organizer: 第40回日本神経科学大会
• [Presentation] Cellular Mechanisms Underlying Status Epilepticus: what do animal models tell us? (2017)
  • Author(s): Fukuda, A.
  • Organizer: The 19th Annual Meeting of Infantile Seizure Society International Symposium on Status Epilepticus in Infants and Children
  • Int'l Joint Research / Invited
• [Presentation] Prenatal stress on Gad1-heterozygotes selectively perturbs parvalbumin (PV)-positive GABAergic neurogenesis, GABA synapses and social interaction behavior. (2017)
  • Author(s): Wang, T., Sinha Adya, S., Yanagawa, Y., Kawai, T., Hata, K. and Fukuda, A.
  • Organizer: IBNS 26th Annual Meeting
  • Int'l Joint Research
• [Remarks] 浜松医科大学医学部神経生理学講座ホームページ
  • URL: http://www.hama-med.ac.jp/uni_education_igakubu_igaku_seiri1.html