2022 Fiscal Year Final Research Report
Gene expression analysis of spermatogonial stem cell niche in undesended testis by single cell RNA-sequence
Project/Area Number |
19K18592
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Research Category |
Grant-in-Aid for Early-Career Scientists
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Allocation Type | Multi-year Fund |
Review Section |
Basic Section 56030:Urology-related
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Research Institution | Nagoya 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)
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Project Period (FY) |
2019-04-01 – 2023-03-31
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Keywords | 精子幹細胞 / 男性不妊症 |
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.
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Free Research Field |
男性不妊症
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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。
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