脂質代謝異常による無精子症発症機構の解明

2016 Fiscal Year Research-status Report

• Project/Area Number: 16K11121
• Research Institution: Tokyo University of Science
• Principal Investigator: 大和屋 健二 東京理科大学, 理工学部応用生物科学科, 助教 (80447309)
• Co-Investigator(Kenkyū-buntansha): 宮戸 健二 国立研究開発法人国立成育医療研究センター, 細胞医療研究部, 室長 (60324844)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 精子形成

Outline of Annual Research Achievements

脂質代謝に関わると思われるタンパク質のコンディショナルノックアウトマウスを作成したところ、Floxedマウスには雌雄共に妊孕性が維持されていることが確認された。Creリコンビナーゼ発現マウスとの交配で得られたコンディショナルノックアウトマウスは雄性赴任であることが認められた。
このノックアウトマウスの形態的解析を主に行った。具体的にはコンディショナルノックアウトマウスとFloxedマウスをブアン固定し、精巣と精巣上体のパラフィン切片を作成し、PAS-hematoxylin染色を行った。その結果、Floxedマウスの精巣上体には成熟精子が認められたが、コンディショナルノックアウトマウスの精巣上体には正常な成熟精子は認められなかった。Floxedマウスの精巣ではすべてのステージが認められたが、コンディショナルノックアウトマウスの精巣切片では、減数分裂中の精母細胞は確認できたが、伸長期精子細胞は認められなかった。現在、異常が認められるステージを明らかにし、透過電子顕微鏡により微細形態の異常を観察を行っている。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

精子形成に異常があり、どのステージで現れるかを絞り込めたため。

Strategy for Future Research Activity

特定のステージの異常が認められたが、セルトリ細胞側か精子細胞側の異常かが問題である。
そこで、電子顕微鏡観察によって微細形態観察を行う。
また、別のプロモーター制御下のCreリコンビナーゼ発現マウスを導入し、セルトリ細胞側か精子細胞側の異常かを明らかにすることを検討したい。

Causes of Carryover

年度内に収まりそうだった実験が延びたため。

Expenditure Plan for Carryover Budget

昨年度から継続して行っている実験に用いる。

Research Products

• [Journal Article] Exosomes versus microexosomes: Shared components but distinct functions (2017)
  • Author(s): Kenji Miyado, Woojin Kang, Kenji Yamatoya, Maito Hanai, Akihiro Nakamura, Toshiyuki Mori, Mami Miyado, Natsuko Kawano
  • Journal Title: JOURNAL OF PLANT RESEARCH Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1007/s10265-017-0907-7
  • Peer Reviewed
• [Journal Article] Increased incidence of post-term delivery and Cesarean section after frozen-thawed embryo transfer during a hormone replacement cycle (2017)
  • Author(s): Kazuki Saito, Kenji Miyado, Kenji Yamatoya, Akira Kuwahara, Eisuke Inoue, Mami Miyado, Maki Fukami, Tomonori Ishikawa, Takakazu Saito, Toshiro Kubota, Hidekazu Saito
  • Journal Title: JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1007/s10815-017-0869-7
  • Peer Reviewed
• [Journal Article] Complementary Critical Functions of Zfy1 and Zfy2 in Mouse Spermatogenesis and Reproduction (2017)
  • Author(s): Takashi Nakasuji, Narumi Ogonuki, Tomoki Chiba, Tomomi Kato, Kumiko Shiozawa, Kenji Yamatoya, Hiromitsu Tanaka, Tadashi Kondo, Kenji Miyado, Naoyuki Miyasaka, Toshiro Kubota, Atsuo Ogura, Hiroshi Asahara
  • Journal Title: PLoS Genetics Volume: - Pages: -
  • DOI: 10.1371/journal.pgen.1006578
  • Peer Reviewed / Open Access
• [Journal Article] 体外受精における人工的操作とヒト成長への影響 (2016)
  • Author(s): 宮戸健二、大和屋健二、齊藤和毅、河野菜摘子、齊藤英和
  • Journal Title: ＨＯＲＭＯＮＥ ＦＲＯＮＴＩＥＲ ＩＮ ＧＹＮＥＣＯＬＯＧＹ Volume: 23 Pages: 167-171
• [Presentation] Function analysis of CD9 and egg exosome-mediated membrane fusion process in fertilization (2016)
  • Author(s): Maito Hanai, Kanako Hayami, Kenji Yamatoya, Woojin Kang, Natsuko Kawano, Kenji Miyado
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Physiological activity of Cistanche and Nitraria extracts in mammals (2016)
  • Author(s): Saki Naohara, Kenji Yamatoya, Yoji Yoshimi , Tomohiko Kato , Kazuya Nakata , Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Exploratory research of physiological activity of newly synthesized benzoic acid derivative having a retinoid activity (2016)
  • Author(s): Yukiko Wada, Kenji Yamatoya, Yoji Yoshimi, Shinichi Saito, Kazuya Nakata, Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Pharmacology of Rare sugars in hypoxic conditions (2016)
  • Author(s): Yuka Takahashi, Kenji Yamatoya, Yoji Yoshimi, Kazuya Nakata, Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Analysis of mitochondrial dysfunction induced by Ridaifen-G in mammalian cells (2016)
  • Author(s): Kousuke Yamada, Kenji Yamatoya, Yoji Yoshimi, Manabu Morita, Takuya Yamaki, Nozomi Ota, Natsumi Shibata, Isamu Shiina, Kazuya Nakata, Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Searching the enzyme responsible for the cleavage of sperm equatorial segment protein 1 in the mouse (2016)
  • Author(s): Kenji Yamatoya, Kazuya Nakata, Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Tamaki Seki, Kenji Yamatoya, Kazuya Nakata, Masahiko Ikekita (2016)
  • Author(s): Searching the enzyme responsible for the cleavage of sperm acrosome membrane-associated protein 1 in the mouse
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research
• [Presentation] Exploration of the new drug substance of the natural compound indicating bioactivity (2016)
  • Author(s): Misaki Kabasawa, Kenji Yamatoya, Koji Kuramochi, Kazuya Nakata, Masahiko Ikekita
  • Organizer: The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan
  • Place of Presentation: Okinawa convention center. Okinawa, (Japan)
  • Year and Date: 2016-11-17 – 2016-11-18
  • Int'l Joint Research