in vivo fluorescence imaging research on the possible role of prostaglandins in modulation of GnRH secretion

• Project/Area Number: 17H03931
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Integrative animal science
• Research Institution: The University of Tokyo
• Principal Investigator: Matsuwaki Takashi 東京大学, 大学院農学生命科学研究科(農学部), 准教授 (20447361)
• Co-Investigator(Kenkyū-buntansha): 角田 茂 東京大学, 大学院農学生命科学研究科(農学部), 准教授 (80345032)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000); Fiscal Year 2019: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
• Keywords: GnRHmニューロン / 神経興奮 / in vivo 脳内蛍光顕微鏡 / プロスタグランジン / GnRHサージ / GnRHニューロン / Cyclooxygenase / GnRH / GCaMP / パルスジェネレーター / サージジェネレーター / LHパルス

Outline of Final Research Achievements

- Using the transgenic rats with GFP expression in GnRH neurons, we have established a novel in vivo fluorescence imaging system for observation of the living GnRH neurons in the deep part of brain. The cell bodies were clearly distinguishable as a single-cell level.
- In wild-type cycling female rats, the expression levels of both COX-1 and COX-2 dropped in the noon of proestrus stage.
- We have generated mPGES-1 KO rats, using CRISPR/Cas12a-gene editing method. The rats lacks splice acceptor site in exon2.

Academic Significance and Societal Importance of the Research Achievements

これまで、COX-1は発現量の変動がない構成型、COX-2は炎症刺激で発現量が増加する誘導型として知られていた。しかし本研究で、雌の発情周期に依存して両酵素の脳内発現量が変動するという新しい知見が得られた。この事実は、これまで不明だったPGsによるGnRH/LH分泌制御機構の解明のための大きな一助となる。
また、本研究で確立したin vivo 蛍光観察法は、実験動物を生かしたままで脳深部を観察することが出来るため、GnRHニューロンに限らず様々な神経細胞活動の解析に広く活用されることが期待される。

Research Products

• [Int'l Joint Research] 国立台湾大学(その他の国・地域)
• [Int'l Joint Research] 国立台湾大学(台湾)
• [Journal Article] Convulsive responses to seizure-inducible drugs are exacerbated in progranulin-deficient mice (2020)
  • Author(s): Kuroda M, Matsuwaki T, Tanaka Y, Yamanouchi K, Nishihara M
  • Journal Title: Neuroreport Volume: 31 Issue: 6 Pages: 478-483
  • DOI: 10.1097/wnr.0000000000001425
  • Peer Reviewed
• [Journal Article] Involvement of interleukin-1 type 1 receptors in lipopolysaccharide-induced sickness responses. (2017)
  • Author(s): Matsuwaki T, Shionoya K, Ihnatko R, Eskilsson A, Kakuta S, Dufour S, Schwaninger M, Waisman A, Muller W, Pinteaux E, Engblom D, Blomqvist A.
  • Journal Title: Brain, behavior, and immunity Volume: 66 Pages: 165-176
  • DOI: 10.1016/j.bbi.2017.06.013
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Immune-Induced Fever Is Dependent on Local But Not Generalized Prostaglandin E2 Synthesis in the Brain. (2017)
  • Author(s): Eskilsson A*, Matsuwaki T*, Shionoya K*, Mirrasekhian E, Zajdel J, Schwaninger M, Engblom D, Blomqvist A.
  • Journal Title: Journal of Neuroscience Volume: 37 Issue: 19 Pages: 5035-5044
  • DOI: 10.1523/jneurosci.3846-16.2017
  • Peer Reviewed / Int'l Joint Research
• [Presentation] i-GONAD法を用いた簡便な遺伝子欠損マウスおよびラット作出の試み (2019)
  • Author(s): 山藤あかり、角田 茂、松脇貴志、大塚正人、久和 茂
  • Organizer: 第162回 日本獣医学会学術集会
• [Presentation] Generation of gene-modified mice and rats by i-GONAD method. (2019)
  • Author(s): Akari Sando, Shigeru Kakuta, Takashi Matsuwaki, Yu Wei Chang, Tetsuhiro Ogawa, Masato Ohtsuka, Shigeru Kyuwa
  • Organizer: 1st ToBeST symposium, Seoul
  • Int'l Joint Research
• [Presentation] The role of the IL-1 receptor in the centrally-elicited sickness response to lipopolysaccharide (2017)
  • Author(s): Matsuwaki T, Shionoya K, Ihnatko R, Eskilsson A, Kakuta S, Dufour S, Schwaninger M, Waisman A, Muller W, Pinteaux E, Engblom D, Blomqvist A.
  • Organizer: International Cytokine & Interferon Society 2017
  • Int'l Joint Research