精細胞の分化を制御するセルトリ細胞の機能的役割の解明

• Project/Area Number: 20H03162
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 42030:Animal life science-related
• Research Institution: The University of Tokyo
• Principal Investigator: 小沢 学 東京大学, 医科学研究所, 准教授 (80608787)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2022: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2021: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2020: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 精子形成 / 精巣 / セルトリ細胞 / 選択的スプライシング / 精細胞 / マウス / ゲノム編集

Outline of Research at the Start

持続的精子形成は、「精原細胞の維持（自己複製）」と「精細胞の分化（減数分裂による半数体精子細胞の産生）」という異なるプロセスがバランスよく厳密に維持されることによって成り立っている。このプロセスには精細管内に存在するセルトリ細胞からのサポートが不可欠であるが、「精細胞の分化」を制御するセルトリ細胞の機能的役割は不明な点が多い。本研究では、これまでその分子機序がほとんど明らかにされていないセルトリ細胞による精細胞の減数分裂制御メカニズムを明らかにすることを目的とする。

Outline of Annual Research Achievements

持続的精子形成は、「精原細胞の維持」と「精細胞の分化」という異なるプロセスがバランスよく厳密に維持されることによって成り立っている。このプロセスには精細管内に存在するセルトリ細胞からのサポートが不可欠であるが、その機能的役割の詳細はこれまでのところ不明な点が多い。申請者らは、選択的スプライシング制御因子であるPTBP1がセルトリ細胞で高発現することに着目し、本課題において、セルトリ細胞特異的なPTBP1欠損マウスを作製してその表現型解析を行った。その結果、精細胞の分化異常が起こり、雄性不妊になることを見出した。また、PTBP1欠損マウスではセルトリ細胞のF-アクチン構造に異常が生じることを明らかにした。更に、セルトリ細胞をセルソーターを用いて単離しPTBP1と結合するRNAを次世代シーケンサーにより網羅的に探索したところ、細胞骨格の維持に関与するシグナル伝達因子が多数検出された。さらに、PTBP1欠損したセルトリ細胞株にこれらの因子を強制発現させることにより、細胞骨格の異常が回復することを確認した。本研究の結果より、PTBP1はセルトリ細胞の細胞骨格の維持を介して精細胞の分化を制御するというモデルが示唆された。

Research Progress Status

令和4年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和4年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Multiple ageing effects on testicular/epididymal germ cells lead to decreased male fertility in mice (2024)
  • Author(s): Tsutomu Endo, Kiyonori Kobayashi, Takafumi Matsumura, Chihiro Emori, Manabu Ozawa, Shimpei Kawamoto, Daisuke Okuzaki, Keisuke Shimada, Haruhiko Miyata, Kentaro Shimada, Mayo Kodani, Yu Ishikawa-Yamauchi, Daisuke Motooka, Eiji Hara, Masahito Ikawa
  • Journal Title: Communications Biology Volume: 7(1) Issue: 1 Pages: 1-6
  • DOI: 10.1038/s42003-023-05685-2
  • Peer Reviewed / Open Access
• [Journal Article] Calcium-binding protein 7 expressed in muscle negatively regulates age-related degeneration of neuromuscular junctions in mice (2024)
  • Author(s): Eguchi Takahiro、Tezuka Tohru、Watanabe Yuji、Inoue-Yamauchi Akane、Sagara Hiroshi、Ozawa Manabu、Yamanashi Yuji
  • Journal Title: iScience Volume: 27 Issue: 2 Pages: 108997-108997
  • DOI: 10.1016/j.isci.2024.108997
  • Peer Reviewed / Open Access
• [Journal Article] Age-related decline in spermatogenic activity accompanied with endothelial cell senescence in male mice_ (2023)
  • Author(s): Manabu Ozawa, Hideto Mori, Tsutomu Endo, Yu Ishikawa-Yamauchi, Daisuke Motooka, Chihiro Emori, Masahiro Ikawa
  • Journal Title: iScience_ Volume: 26(12) Issue: 12 Pages: 108456-108456
  • DOI: 10.1016/j.isci.2023.108456
  • Peer Reviewed / Open Access
• [Journal Article] TLR7/8 stress response drives histiocytosis in SLC29A3 disorders (2023)
  • Author(s): Shibata Takuma、Sato Ryota、Taoka Masato、Saitoh Shin-Ichiroh、Komine Mayumi、（他16人）、Ohto Umeharu、Shimizu Toshiyuki、Ozawa Manabu、Yoshida Nobuaki、Isobe Toshiaki、Latz Eicke、Mukai Kojiro、Taguchi Tomohiko、Hemmi Hiroaki、Akira Shizuo、Miyake Kensuke
  • Journal Title: Journal of Experimental Medicine Volume: 220 Issue: 9 Pages: 320230054-320230054
  • DOI: 10.1084/jem.20230054
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] LONRF2 is a protein quality control ubiquitin ligase whose deficiency causes late-onset neurological deficits (2023)
  • Author(s): Li Dan、Johmura Yoshikazu、Morimoto Satoru et al.
  • Journal Title: Nature Aging Volume: 3 Issue: 8 Pages: 1001-1019
  • DOI: 10.1038/s43587-023-00464-4
  • Peer Reviewed / Open Access
• [Journal Article] Ptbp1 deletion does not induce astrocyte-to-neuron conversion (2023)
  • Author(s): Hoang Thanh、Kim Dong Won、Appel Haley、Ozawa Manabu、Zheng Sika、Kim Juhyun、Blackshaw Seth
  • Journal Title: Nature Volume: 618 Issue: 7964 Pages: E1-E7
  • DOI: 10.1038/s41586-023-06066-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models (2022)
  • Author(s): Ozawa Manabu、Emori Chihiro、Ikawa Masahito
  • Journal Title: Journal of Visualized Experiments Volume: 186 Issue: 186 Pages: 1-13
  • DOI: 10.3791/64385
  • Peer Reviewed / Open Access
• [Journal Article] Efficient simultaneous double DNA knock-in in murine embryonic stem cells by CRISPR/Cas9 ribonucleoprotein-mediated circular plasmid targeting for generating gene-manipulated mice (2022)
  • Author(s): Ozawa Manabu、Taguchi Jumpei、Katsuma Kento、Ishikawa-Yamauchi Yu、Kikuchi Mio、Sakamoto Reiko、Yamada Yasuhiro、Ikawa Masahito
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 21558-21558
  • DOI: 10.1038/s41598-022-26107-z
  • Peer Reviewed / Open Access
• [Journal Article] Splice factor polypyrimidine tract-binding protein 1 (Ptbp1) primes endothelial inflammation in atherogenic disturbed flow conditions (2022)
  • Author(s): Hensel Jessica A.、Nicholas Sarah-Anne E.、Kimble Amy L.、Nagpal Arjun S.、Omar Omar M. F.、Tyburski Jordan D.、Jellison Evan R.、Menoret Antoine、Ozawa Manabu、Rodriguez-Oquendo Annabelle、Vella Anthony T.、Murphy Patrick A.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 119 Issue: 30
  • DOI: 10.1073/pnas.2122227119
  • Peer Reviewed / Open Access
• [Journal Article] Genetic loss of function of Ptbp1 does not induce glia-to-neuron conversion in retina (2022)
  • Author(s): Hoang Thanh、Kim Dong Won、Appel Haley、Pannullo Nicole A.、Leavey Patrick、Ozawa Manabu、Zheng Sika、Yu Minzhong、Peachey Neal S.、Blackshaw Seth
  • Journal Title: Cell Reports Volume: 39 Issue: 11 Pages: 110849-110849
  • DOI: 10.1016/j.celrep.2022.110849
  • Peer Reviewed / Open Access
• [Journal Article] MYCL-mediated reprogramming expands pancreatic insulin-producing cells (2022)
  • Author(s): Hirano Michitada、So Yusei、Tsunekawa Shin、Kabata Mio、Ohta Sho、Sagara Hiroshi、Sankoda Nao、Taguchi Jumpei、Yamada Yosuke、Ukai Tomoyo、Kato Makoto、Nakamura Jiro、Ozawa Manabu、Yamamoto Takuya、Yamada Yasuhiro
  • Journal Title: Nature Metabolism Volume: 4 Issue: 2 Pages: 254-268
  • DOI: 10.1038/s42255-022-00530-y
  • Peer Reviewed / Open Access
• [Journal Article] Generation of mice for evaluating endogenous p16Ink4a protein expression (2022)
  • Author(s): Shimada-Takayama Yui、Yasuda Takayuki、Ukai Tomoyo、Taguchi Jumpei、Ozawa Manabu、Sankoda Nao、Ohta Sho、Yamada Yasuhiro
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 599 Pages: 43-50
  • DOI: 10.1016/j.bbrc.2022.02.005
  • Peer Reviewed / Open Access
• [Journal Article] Skewed endosomal RNA responses from TLR7 to TLR3 in RNase T2-deficient macrophages (2021)
  • Author(s): Liu Kaiwen、Sato Ryota、Shibata Takuma、Hiranuma Ryosuke、Reuter Tatjana、Fukui Ryutaro、Zhang Yun、Ichinohe Takeshi、Ozawa Manabu、Yoshida Nobuaki、Latz Eicke、Miyake Kensuke
  • Journal Title: International Immunology Volume: 33 Issue: 9 Pages: 479-490
  • DOI: 10.1093/intimm/dxab033
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] DMRT1-mediated reprogramming drives development of cancer resembling human germ cell tumors with features of totipotency (2021)
  • Author(s): Taguchi Jumpei、Shibata Hirofumi、Kabata Mio、Kato Masaki、Fukuda Kei、Tanaka Akito、Ohta Sho、Ukai Tomoyo、Mitsunaga Kanae、Yamada Yosuke、Nagaoka So I、Yamazawa Sho、Ohnishi Kotaro、Woltjen Knut、Ushiku Tetsuo、Ozawa Manabu、Saitou Mitinori、Shinkai Yoichi、Yamamoto Takuya、Yamada Yasuhiro
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 5041-5041
  • DOI: 10.1038/s41467-021-25249-4
  • Peer Reviewed / Open Access
• [Journal Article] NELL2-mediated lumicrine signaling through OVCH2 is required for male fertility. Science. (2020)
  • Author(s): Kiyozumi D, Noda T, Yamaguchi R, Tobita T, Matsumura T, Shimada K, Kodani M, Kohda T, Fujihara Y, Ozawa M, Yu Z, Miklossy G, Bohren KM, Horie M, Okabe M, Matzuk MM, Ikawa M.
  • Journal Title: Science. Volume: 368 Issue: 6495 Pages: 1132-1135
  • DOI: 10.1126/science.aay5134
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of a p16 Reporter Mouse and Its Use to Characterize and Target p16high Cells In Vivo. (2020)
  • Author(s): Omori S, Wang TW, Johmura Y, ..., Sakamoto T (15番目), ..., Nakanishi M.
  • Journal Title: Cell Metab Volume: 32 Issue: 5 Pages: 814-828
  • DOI: 10.1016/j.cmet.2020.09.006
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Declining spermatogenic activities accompanied by the senescent feature of endothelial cells in male mice with age (2023)
  • Author(s): Manabu Ozawa, Hideto Mori, Tsutomu Endo, Masahito Ikawa
  • Organizer: Society for the Study of Reproduction
  • Int'l Joint Research
• [Book] Genome Editing in Animals (2023)
  • Author(s): Manabu Ozawa, Chihiro Emori, Masahito Ikawa
  • Total Pages: 399
  • Publisher: Springer
• [Patent(Industrial Property Rights)] 超長鎖DNAの効率的なノックインによるヒト化マウス作製技術の開発 (2023)
  • Inventor(s): 小沢学、田口純平
  • Industrial Property Rights Holder: 小沢学、田口純平
  • Industrial Property Rights Type: 特許
  • Filing Date: 2023