メダカ脳におけるメタスチンニューロンの新規生理機能に関する研究

• Project/Area Number: 08F08089
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Animal physiology/Animal behavior
• Research Institution: The University of Tokyo
• Principal Investigator: 岡 良隆 The University of Tokyo, 大学院・理学系研究科, 教授
• Co-Investigator(Kenkyū-buntansha): MD. Sadiqul 東京大学, 大学院・理学系研究科, 外国人特別研究員 ; SADIQUL Md. 東京大学, 大学院・理学系研究科, 外国人特別研究員
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2009: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2008: ¥800,000 (Direct Cost: ¥800,000)
• Keywords: 神経科学 / 生理学 / 脳・神経 / 動物 / 生理活性 / ペプチド / GnRH / キスペプチン

Research Abstract

GnRH2ニューロンとGnRH3ニューロンにEGFP蛍光タンパク質を発現するトランスジェニックメダカの作成に成功し、蛍光標識および抗GFP抗体を用いた免疫組織化学によりメダカGnRH2およびGnRH3神経系の全体像が把握できた。さらに、酸化型では無毒なMetronidazolを還元することによってDNAに架橋構造を造って細胞毒性を持つようになる還元酵素Nitroreductase(NTR)タンパク質をGnRHニューロンに遺伝子導入したトランスジェニックメダカの作成を開始した。遺伝子コンストラクトは完成し、既に受精卵への微量注射を行っている。現在、遺伝子導入されたメダカをスクリーニングして導入遺伝子をホモでもつトランスジェニック系統を確立しつつある。トランスジェニックメダカの系統が完成すれば、これを用いてGnRH1,2,3ニューロンを特異的に細胞破壊し、メタスチンニューロンとの相互関係や行動、生殖などがどのように変化するかを生理学的に解析することができる。機能とアミノ酸配列の異なる3種類のGnRHニューロンそれぞれにGFP蛍光タンパク質を発現させるトランスジェニックメダカは既に当研究室で作成している。この遺伝子コンストラクトを参考にして、成魚まで透明性を保つ透明メダカ系統のGnRHニューロンおよびメタスチンニューロンにGFP等の蛍光タンパク質のみならずKRやNTRを遺伝子導入したトランスジェニックメダカの作成に成功すれば、それらのニューロンを細胞特異的に破壊し、生理・行動実験によりそれぞれのニューロン間の相互作用を解析する事ができるようになり、脊椎動物における生殖の中枢制御の本質的理解につながる知見が得られる。

Research Products

• [Journal Article] Regular pacemaker activity characterizes GnRH 2 neurons recorded from GFP°transgenic medaka. (2010)
  • Author(s): Kanda, S., Nishikawa, K., Karigo, T., Okubo, K., Isomae, S, Abe, H., Kobayashi, D., Oka, Y.
  • Journal Title: Endocrinology 151 Pages: 695-701
  • Peer Reviewed
• [Journal Article] Hypothalamic kiss1 but not kiss2 neurons are involved in estrogen feedback in medaka (Oryzias latipes) (2010)
  • Author(s): Mitani, Y., Kanda, S., Akazome, Y., Zempo, B., Oka, Y.
  • Journal Title: Endocrinology 151 Pages: 1751-1759
  • Peer Reviewed
• [Journal Article] Excitatory action of GABA in the terminal nerve gonadotropin-releasing hormone neurons. (2010)
  • Author(s): Nakane, R., Oka, Y.
  • Journal Title: Journal of Neurophysiology 103 Pages: 1375-1384
  • Peer Reviewed
• [Journal Article] Functional and evolutionary insights into vertebrate kisspeptin systems from studies of fish brain. (2010)
  • Author(s): Akazome, Y., Kanda, S., Okubo, K., Oka, Y.
  • Journal Title: Journal of Fish Biology, Special Issue "Reproductive Physiology" 76 Pages: 161-182
  • Peer Reviewed
• [Journal Article] Primary culture of the isolated terminal nerve-gonadotrophin releasing hormone (GnRH) neurones derived from adult teleost(dwarf gourami, Colisa lalia) brain for the study of peptide release mechanisms (2009)
  • Author(s): Abe, H., Oka, Y.
  • Journal Title: Journal of Neuroendocrinology 21 Pages: 489-505
  • Peer Reviewed
• [Journal Article] Possible role of oestrogen in pubertal increase of Kiss1/Kisspeptin expression in discrete hypothalamic areas of female rats. (2009)
  • Author(s): Takase, K., Uenoyama, Y., Inoue, N., Matsui, H., Yamada, S., Shimizu, M., Homma, T., Toniikawa, J., Kanda, S., Matsumoto, H., Oka, Y., Tsukamura, H., Maeda, K.-I.
  • Journal Title: Journal of Neuroendocrinology 21 Pages: 527-537
  • Peer Reviewed
• [Journal Article] Three types of gonadotrophin-releasing hormone neurones and steroid-sensitive sexually dimorphic kisspeptin neurones in teleosts. (2009)
  • Author(s): Oka, Y.
  • Journal Title: Journal of Neuroendocrinology 21 Pages: 334-338
  • Peer Reviewed
• [Journal Article] Primary culture of the isolated terminal nerve-gonadotrophin releasing hormone (GnRH) neurones derived from adult teleost (dwarf gourami, Colisa lalia) brain for the study of peptide release mechanisms (2009)
  • Author(s): Abe, H., Oka, Y.
  • Journal Title: Journal of Neuroendocrinology 21 Pages: 489-505
  • Peer Reviewed
• [Journal Article] Three types of gonadotrophin-releasing hormone neurones and steroid-sensitive sexually dimorphic kisspeptin neurones in teleosts (2009)
  • Author(s): Oka, Y.
  • Journal Title: Journal of Neuroendocrinology 21 Pages: 334-338
  • Peer Reviewed
• [Journal Article] Possible role of oestrogen in pubertal increase of Kiss1/Kisspeptin expression in discrete hypothalamic areas of female rats (2009)
  • Author(s): Takase, K., Uenoyama, Y., Inoue, N., Matsui, H., Yamada, S., Shimizu, M., Homma, T., Tomikawa, J., Kanda, S., Matsumoto, H., Oka, Y., Tsukamura, H., Maeda, K.-I.
  • Journal Title: Journal of Neuroendocrinology ((印刷中)掲載確定)
  • Peer Reviewed
• [Journal Article] Identification of KiSS-1 product kisspeptin and steroid-sensitive sexually-dimorphic kisspeptin neurons in medaka (Oryzias latipes) (2008)
  • Author(s): Kanda, S., Akazome, Y., Yamamoto, N., Yamada, S., Tsukamura, H., Maeda, K., Oka, Y.
  • Journal Title: Endocrinology 149 Pages: 2467-2476
  • Peer Reviewed
• [Journal Article] Coordinated synchronization in the electrically coupled network of terminal nerve gonadotropin-releasing hormone neurons as demonstrated by double patch-clamp study (2008)
  • Author(s): Haneda, K., Oka, Y.
  • Journal Title: Endocrinology 149 Pages: 3540-3548
  • Peer Reviewed
• [Journal Article] Immunohistochemical localization of GnRH-like peptide in the brain of the cephalopod spear squid, Loligo bleekeri (2008)
  • Author(s): Amano, M., Oka, Y., Nagai, Y., Amiya, N., Yamamori, K.
  • Journal Title: General and Comparative Endocrinology 156 Pages: 277-284
  • Peer Reviewed
• [Journal Article] Identification and expression analysis of peroxisome proliferator-activated receptors cDNA in a reptile, the leopard gecko (Eublepharis macularius) (2008)
  • Author(s): Kato, K., Oka, Y., Park, M. K.
  • Journal Title: Zoological Science 25 Pages: 492-502
  • Peer Reviewed
• [Presentation] Generation of transgenic medaka expressing bacterial nitroreductase (NTR) in GnRH2 and GnRH3 neurons (2010)
  • Author(s): Sadiqul, MD.Okubo, K., Takahashi, A., Kanda, S., Akazome, Y., Oka, Y.
  • Organizer: 日本動物学会関東支部大会
  • Place of Presentation: つくば
  • Year and Date: 2010-03-13
• [Presentation] 単一細胞電気穿孔法を用いて外来遺伝子導入した終神経GnRHニューロンにおける分泌小胞の可視化 (2009)
  • Author(s): 阿部秀樹、入江高行、岡良隆
  • Organizer: 第32回日本神経科学大会
  • Place of Presentation: 名古屋
• [Presentation] キンギョ嗅球神経回路におけるGnRHペプチドの神経修飾作用 (2009)
  • Author(s): 河合喬文、阿部秀樹、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] メダカにおける視床下部GnRHニューロン活動と下垂体遺伝子発現の日内変動 (2009)
  • Author(s): 苅郷友美、神田真司、相川雅人、阿部秀樹、大久保範聡、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] メダカ脳におけるkiss1およびkiss2ニューロンの生殖制御に対する関与 (2009)
  • Author(s): 三谷優太、赤染康久、神田真司、善方文太郎、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] メダカキスペプチン神経系による視索前野GnRH神経系の制御 (2009)
  • Author(s): 神田真司、赤染康久、大久保範聡、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] GFPトランスジェニックメダカを用いたGnRH2ニューロンの生理学的解析 (2009)
  • Author(s): 西川圭、神田真司、阿部秀樹、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] メダカGnRH3ニューロンにおけるco-transmitterとしてのグルタミン酸 (2009)
  • Author(s): 赤染康久、神田真司、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] 終神経GnRHニューロンのペースメーカー活動に対するGABAの興奮性作用 (2009)
  • Author(s): 中根亮、岡良隆
  • Organizer: 日本動物学会第80回大会
  • Place of Presentation: 静岡
• [Presentation] Kisspeptin neurons act closely but indirectly on GnRH 1 neuronsvia local interneurons but not on GnRH 2 or 3 neurons in medaka (2009)
  • Author(s): Kanda, S., Akazome, Y., Okubo, K., Okamura, H., Oka, Y.
  • Organizer: The 39th Annual Meeting of the Society for Neuroscience 2009
  • Place of Presentation: Chicago, U.S.A.
• [Presentation] Diurnal regulation of GnRH1 neuronal activities and gonadotropin subunit transcription in a daily spawning fish, medaka (2009)
  • Author(s): Karigo, T., Kanda, S., Aikawa, M., Abe, H., Okubo, K., Oka, Y.
  • Organizer: The 39th Annual Meeting of the Society for Neuroscience 2009
  • Place of Presentation: Chicago, U.S.A.
• [Presentation] Visualization of peptidergic dense cored vesicles in the terminal nerve (TN)-gonadotropin releasing hormone(GnRH) neurons by single cell electroporation of exogenous genes (2009)
  • Author(s): Abe, H., Irie, T., Oka, Y.
  • Organizer: The 39th Annual Meeting of the Society for Neuroscience 2009
  • Place of Presentation: Chicago, U.S.A.
• [Presentation] Electrophysiological, anatomical, and molecular characterization of GnRH neurons in dwarf gourami and GFP transgenic medaka (2009)
  • Author(s): Oka, Y.
  • Organizer: The 36th International Congress of Physiological Sciences
  • Place of Presentation: Kyoto
• [Presentation] Electrophysiological and anatomical characterization of three distinct GnRH neuronal populations in the GFP transgenic medaka (2008)
  • Author(s): Kanda, S., Okubo, K., Isomae, S., Abe, H., Kobayashi, D., Takeda, H., Aida, K., Nagahama, Y., Oka, Y.
  • Organizer: Annual Meeting Society for Neuroscience 2008
  • Place of Presentation: Washington DC, U.S.A.
  • Year and Date: 2008-11-17
• [Presentation] Primary culture of the isolated terminal nerve-gonadotropin releasing hormone neurons derived from adult teleost brain and their electrophysiological properties (2008)
  • Author(s): Abe, H., Oka, Y.
  • Organizer: Annual Meeting Society for Neuroscience 2008
  • Place of Presentation: Washington DC, U.S.A.
  • Year and Date: 2008-11-17
• [Presentation] Calcium oscillation of the olfactory non-sensory cells in the goldfish (2008)
  • Author(s): Kawai, T., Abe, H., Oka, Y.
  • Organizer: Annual Meeting Society for Neuroscience 2008
  • Place of Presentation: Washington DC, U.S.A.
  • Year and Date: 2008-11-17
• [Presentation] Coordinated synchronization in the electrically coupled network of terminal nerve GnRH (gonadotropin-releasing hormone) neurons as demonstrated by double patch-clamp study (2008)
  • Author(s): Oka, Y., Haneda, K.
  • Organizer: Annual Meeting Society for Neuroscience 2008
  • Place of Presentation: Washington DC, U.S.A.
  • Year and Date: 2008-11-17
• [Presentation] 脊椎動物に共通する生殖の中枢制御のしくみをメダカの脳から知る (2008)
  • Author(s): 岡良隆
  • Organizer: 第101回日本繁殖生物学会シンポジウム「新規生理活性因子メタスチン」
  • Place of Presentation: 福岡
  • Year and Date: 2008-09-20
• [Presentation] Medaka brain teaches us the mechanisms of the central regulation of reproduction common to the vertebrate species (2008)
  • Author(s): Oka, Y.
  • Organizer: 第35回日本神経内分泌学会・第23回日本下垂体研究会シンポジウム「視床下部・下垂体調節機構の新展開」
  • Place of Presentation: 東京
  • Year and Date: 2008-08-28
• [Presentation] Evolutionary aspects of peptidergic nervous systems in the vertebrate brain (2008)
  • Author(s): Oka, Y.
  • Organizer: International Seminar : Evolutionary Studies in Behavioral Neuroscience
  • Place of Presentation: Yokosuka
• [Presentation] Functional analysis of the physiology of peptidergic systems using the transgenic medaka" Neurobiology of Animal Behavior and Communication (2008)
  • Author(s): Oka, Y.
  • Organizer: 31st Annual meeting of the Japan Neuroscience Society
  • Place of Presentation: Tokyo
• [Presentation] GABAergic excitation in the terminal nerve GnRH neurons (2008)
  • Author(s): Nakane, R., Oka, Y.
  • Organizer: 第31回日本神経科学大会
  • Place of Presentation: 東京
• [Presentation] 終神経GnRHニューロンへのFMRFamide ペプチドの過分極性作用に関する電気生理学的解析 (2008)
  • Author(s): 齊藤健, 阿部秀樹, 岡良隆
  • Organizer: 日本動物学会第79回大会
  • Place of Presentation: 福岡
• [Presentation] 単一細胞電気穿孔による終神経GnRHニューロンへの外来分子導入法 (2008)
  • Author(s): 入江高行, 尾崎令, 阿部秀樹, 岡良隆
  • Organizer: 日本動物学会第79回大会
  • Place of Presentation: 福岡
• [Presentation] キンギョ嗅上皮におけるカルシウム振動 (2008)
  • Author(s): 河合喬文, 阿部秀樹, 岡良隆
  • Organizer: 日本動物学会第79回大会
  • Place of Presentation: 福岡
• [Presentation] メダカ脳内kiss1遺伝子発現は卵巣エストロゲンにより調節され生殖状態を反映する (2008)
  • Author(s): 神田真司, 赤染康久, 松永拓也, 山本直之, 山田俊児, 束村博子, 前多敬一郎, 岡良隆
  • Organizer: 日本動物学会第79回大会
  • Place of Presentation: 福岡
• [Presentation] Three types of GnRH neurons and steroid-sensitive sexually dimorphic kisspeptin neurons in teleosts (2008)
  • Author(s): Oka, Y.
  • Organizer: US/Japan Neurosteroid Symposium 2008
  • Place of Presentation: Gifu
• [Presentation] Distribution of kisspeptin neurons in the CNS, with attention also to comparative aspects of Kiss1/GPR54 system (2008)
  • Author(s): Vaudry, H., Oka, Y.
  • Organizer: 1st world conference on kisspeptin signaling in the brain
  • Place of Presentation: Cordoba, Spain
• [Presentation] Primary culture system of fish GnRH neurons for the cellular physiological studies of effects of kisspeptin (2008)
  • Author(s): Abe, H., Oka, Y.
  • Organizer: 1st world conference on kisspeptin signaling in the brain
  • Place of Presentation: Cordoba, Spain
• [Presentation] Quantitative expression profiling of kiss1 and its receptor in the teleost medaka (2008)
  • Author(s): Okubo, K., Kanda, S., Oka, Y.
  • Organizer: 1st world conference on kisspeptin signaling in the brain
  • Place of Presentation: Cordoba, Spain
• [Presentation] Possible novel form of kisspeptin and kisspeptin receptors in the medaka, Oryzias latipes (2008)
  • Author(s): Akazome, Y., Kanda, S., Mitani, Y., Okubo, K., Oka, Y.
  • Organizer: 1st world conference on kisspeptin signaling in the brain
  • Place of Presentation: Cordoba, Spain
• [Presentation] Kisspeptin and steroid-sensitive sexually dimorphic kisspeptin neurons in medaka (Oryzias latipes) (2008)
  • Author(s): Kanda, S., Akazome, Y., Yamamoto, N., Yamada, S., Tsukamura, H., Maeda, K., Oka, Y.
  • Organizer: 1st world conference on kisspeptin signaling in the brain
  • Place of Presentation: Cordoba, Spain
• [Remarks]
  • URL: http://www.biol.s.u-tokyo.ac.jp/users/naibunpi/Oka/Oka_lab_publications.html
• [Remarks]
  • URL: http://www.biol.s.u-tokyo.ac.jp/users/naibunpi/Oka/Oka_lab_publications.html