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Gene expression analysis of spermatogonial stem cell niche in undesended testis by single cell RNA-sequence

Research Project

Project/Area Number 19K18592
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 56030:Urology-related
Research InstitutionNagoya City University

Principal Investigator

Moritoki Yoshinobu  名古屋市立大学, 医薬学総合研究院(医学), 研究員 (50595395)

Project Period (FY) 2019-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000)
Fiscal Year 2021: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords精子幹細胞 / 男性不妊症 / 停留精巣 / 1細胞RNAシークエンス / 精子感細胞 / RNA-sequence / RNA-シークエンス
Outline of Research at the Start

本研究では男性不妊症のモデルとして停留精巣を用い、精子幹細胞ニッチの構成細胞ごとの遺伝子発現を下降精巣(正常精巣)と1細胞RNA-sequenceを用いて比較する。本研究により、停留精巣の幹細胞ニッチで発現が変化している遺伝子(群)を1細胞レベルで同定し、停留精巣における男性不妊症の原因を究明する。
具体的には、I:停留精巣モデルマウスの確率、II:TypeB精原細胞、2.マウスES細胞でのRNA-sequence、III:マウス精巣を用いたRNA-sequenceの3段階で行うこととする。

Outline of Final Research Achievements

[Development of a mouse model for undescended testes] Even when flutamide was administered to pregnant mice following the conventional rat technique, it was difficult to create undescended testes. Therefore, we performed surgery to create undescended testes in 3-week-old mice, which correspond to prepubertal age. A small skin incision was performed, and the inguinal canal was ligated and sutured under direct vision to robustly create a mouse model for undesended testes. [Amplification of small amounts of mRNA] cDNA was synthesized and amplified from 10 pg mRNA of GC-1 cells (type B spermatogonia) , equivalent to a single cell, by two methods (SC3-seq vs. Quartz-seq). Amplification efficiency was compared by qPCR. SC3 seq achieved 80% of reproducibility.

Academic Significance and Societal Importance of the Research Achievements

近年、不妊症のカップルが全体の10-15%と増加しており、そのうち約50%に男性側の要因が関与している。男性不妊症の70-75%は、染色体や内分泌学的・解剖学的に異常がない原因不明の特発性であり、メカニズムの全体像は明らかになっていない。
生殖補助医療(ART)の発展に伴い、精子が存在しさえすれば、その運動能や受精能の障壁は乗り越えることができる。しかし精子形成の前段階である精子幹細胞の生存・分化障害を克服する術は、現時点では存在しない。これらの克服には、幹細胞の生存や正常な減数分裂のメカニズムを解明する必要がある。そのため本研究ではまず、幹細胞の維持や精子までの分化に必要な分子を同定する。

Report

(5 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (5 results)

All 2020

All Journal Article (3 results) (of which Peer Reviewed: 3 results) Presentation (2 results)

  • [Journal Article] Disorganization of claudin-11 and dysfunction of the blood-testis barrier during puberty in a cryptorchid rat model2020

    • Author(s)
      Kato Taiki、Mizuno Kentaro、Nishio Hidenori、Moritoki Yoshinobu、Kamisawa Hideyuki、Kurokawa Satoshi、Nakane Akihiro、Maruyama Tetsuji、Ando Ryosuke、Hayashi Yutaro、Yasui Takahiro
    • Journal Title

      Andrology

      Volume: - Issue: 5 Pages: 231-235

    • DOI

      10.1111/andr.12788

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Initial treatment outcome and feasibility of low-dose cabazitaxel against docetaxel- and castration-resistant prostate cancer in a Japanese hospital2020

    • Author(s)
      Chaya Ryosuke, Okamura Takehiko, Yanase Takahiro, Nagai Takashi, Moritoki Yoshinobu, Kobayashi Daichi, Akita Hidetoshi, Yasui Takahiro
    • Journal Title

      Journal of Rural Medicine

      Volume: 15 Issue: 1 Pages: 25-28

    • DOI

      10.2185/jrm.2019-004

    • NAID

      130007789646

    • ISSN
      1880-487X, 1880-4888
    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Myocarditis and myasthenia gravis by combined nivolumab and ipilimumab immunotherapy for renal cell carcinoma: A case report of successful management2020

    • Author(s)
      Yanase Takahiro, Moritoki Yoshinobu, Kondo Hajime, Ueyama Daigo, Akita Hidetoshi, Yasui Takahiro
    • Journal Title

      Urology Case Reports

      Volume: 34 Pages: 101508-101508

    • DOI

      10.1016/j.eucr.2020.101508

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] 2mm 針型鉗子を使用したReduced Port Surgeryによる尿膜管摘除術の初期経験2020

    • Author(s)
      守時 良演、秋田 英俊、柳瀬 貴弘、杉野 輝明、金本 一洋、安井 孝周
    • Organizer
      第34回日本泌尿器内視鏡学会総会
    • Related Report
      2020 Research-status Report
  • [Presentation] 尿管皮膚瘻造設術後の尿管カテーテル選択の検討~シリコンカテーテルとポリウレタンカテーテルの比較~2020

    • Author(s)
      守時 良演、柳瀬 貴弘、杉野 輝明、小林大地、岡村 武彦、秋田 英俊、安井 孝周
    • Organizer
      第108回日本泌尿器科学会総会
    • Related Report
      2020 Research-status Report

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Published: 2019-04-18   Modified: 2024-01-30  

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