Analysis of direct and indirect regulation of kisspeptin neurons on isotocin/vasotocin neurons

2016 Fiscal Year Annual Research Report

• Project/Area Number: 26840111
• Research Institution: The University of Tokyo
• Principal Investigator: 神田 真司 東京大学, 大学院理学系研究科(理学部), 准教授 (50634284)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: キスペプチン / 神経ペプチド

Outline of Annual Research Achievements

本年度は、キスペプチンとバソトシンニューロンの生理学的・形態学的解析を進めることに加え、樹立したキスペプチンノックアウトメダカ、およびバソトシンノックアウトメダカの全脳を用い、RNAseq解析を行った。全脳から精製したmRNAを元に、市販のRNAseqキットを用いてテンプレートを作成、バーコードでサンプルを区別する方法を用い、HiSeq1500でシーケンスを行った。さらに、メダカゲノムデータベースより作成したトランスクリプトに対して、マッピングを行った。この方法で、脳内に存在するmRNAのリード数を算出し、各種キスペプチン遺伝子ノックアウト、バソトシンノックアウトの全脳が発現するmRNAを比較した。そうしたところ、キスペプチンノックアウト、バソトシンノックアウト両方に関連する遺伝子の変動が見られたり、あるいは、キスペプチンノックアウトのみ、バソトシンノックアウトのみにみられる発現変動も見られた。現在それらをreal time PCR法を用いて、精査中である。すでに、一部のものに関しては、real time PCRにおいても再現性よく同じ傾向が見られており、今回行ったRNAseq解析は、正しく網羅的に解析できたと考えている。この中でも、各種神経ペプチドの発現変動がみられたので、それらのノックアウト個体を作成し、さらなる研究を進めている。
なお、前年度までに行ったCa2+イメージングによるキスペプチン神経系によるバソトシン神経系の支配の解析については、追加実験を行った。

Research Products

• [Journal Article] Evolution of the hypothalamic-pituitary-gonadal axis regulation in vertebrates revealed by knockout medaka. (2016)
  • Author(s): Takahashi, A., Kanda, S., Abe, T., and Oka, Y.
  • Journal Title: Endocrinology Volume: 157 Pages: 3994-4002
  • DOI: 10.1210/en.2016-1356
  • Peer Reviewed
• [Journal Article] Female specific glucose-sensitivity of GnRH1 neurons leads to sexually dimorphic inhibition of reproduction in medaka. (2016)
  • Author(s): Hasebe, M., Kanda, S., and Oka, Y.
  • Journal Title: Endocrinology Volume: 157 Pages: 4318-4329
  • DOI: 10.1210/en.2016-1352
  • Peer Reviewed
• [Presentation] Analysis of estrogen feedback mechanism of reproductive regulation in teleosts by using estrogen receprtor knockout female medaka (2016)
  • Author(s): Kayo, D., Kanda, S., Zempo, B., Oka, Y.
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa
  • Year and Date: 2016-11-14 – 2016-11-19
  • Int'l Joint Research
• [Presentation] The kisspeptin neuronal system regulates vasotocin/isotocin neurons via release of neuropeptide B from vPOA Gpr54-1-expressing neurons (2016)
  • Author(s): Nakajo, M., Kanda, S., Karigo, T., and Oka, Y.
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa
  • Year and Date: 2016-11-14 – 2016-11-19
  • Int'l Joint Research
• [Presentation] Electrophysiological and genetic demonstration that VT neuron is critical for water excretion in hypoosmotic conditions in medaka (2016)
  • Author(s): Shinya, M., Kanda, S., Takahashi, A., Konno, N., Hyodo, S., Kagawa, N., and Oka, Y.
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa
  • Year and Date: 2016-11-14 – 2016-11-19
  • Int'l Joint Research
• [Presentation] Kisspeptin may be one of the evolutionary choices specific to mammals for the promotion of folliculogenesis (2016)
  • Author(s): Kanda, S., Arai, Y., Takahashi, A., Abe, T., and Oka, Y.
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa
  • Year and Date: 2016-11-14 – 2016-11-19
  • Int'l Joint Research
• [Presentation] Characiform GnRH neuronal systems provides a good model for functional compensation by paralogous genes (2016)
  • Author(s): Ishida, M., Yang, C., Akazome, Y., Oka, Y., Kanda, S.
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa
  • Year and Date: 2016-11-14 – 2016-11-19
  • Int'l Joint Research
• [Presentation] GnRH receptor in LH cell is a critical red/green signal for LH surge (2016)
  • Author(s): Kanda, S., Arai, Y., Takahashi, A., Abe, T., and Oka, Y.
  • Organizer: The 8th International Symposium on Fish Endocrinology
  • Place of Presentation: Lindholmen Conference Centre, Gothenburg, Sweden
  • Year and Date: 2016-06-28 – 2016-07-02
  • Int'l Joint Research
• [Presentation] Sex steroid-sensitive kisspeptin regulation of various endocrine systems (2016)
  • Author(s): Kanda, S., Nakajo, M., Karigo, T., Oka, Y.
  • Organizer: The 8th Congress of the Asia and Oceania Society for Comparative Endocrinology (AOSCE)
  • Place of Presentation: Korea University College of Medicine, Seoul, Korea
  • Year and Date: 2016-06-20 – 2016-06-24
  • Int'l Joint Research
• [Presentation] Female specific glucose-sensing of GnRH neurons may cause the sexually dimorphic reproductive regulation in malnutritional states (2016)
  • Author(s): Hasebe, M., Kanda, S., Oka, Y.
  • Organizer: The 8th Congress of the Asia and Oceania Society for Comparative Endocrinology (AOSCE)
  • Place of Presentation: Korea University College of Medicine, Seoul, Korea
  • Year and Date: 2016-06-20 – 2016-06-24
  • Int'l Joint Research