Analysis of the interaction between cytoskeleton and Fyn tyrosine kinase during spermatogenesis in the mouse
| Project/Area Number |
10670007
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Chiba University |
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Principal Investigator |
MAEKAWA Mamiko Chiba University, School of Medicine, Assistant, 医学部, 助手 (20181571)
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| Co-Investigator(Kenkyū-buntansha) |
TOYAMA Yoshiro Chiba University, School of Medicine, Lecturer, 医学部, 講師 (70009637)
TOYOTA Fumie Chiba University, School of Medicine, Associate Professor, 医学部, 助教授 (60009751)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Fyn tyrosine kinase / testis / Setoli cell / cytoskeleton / アクチンフィラメント |
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| Research Abstract |
1. Localization of Fyn tyrosine kinase in the mouse testis
Immunohistochemical studies revealed that Fyn tyrosine kinase was localized in the Sertoli cells in the mouse testis, and not in the spermatogenic cells.
2. Colocalization of Fyn and actin filaments in the ectoplasmic specialization
Fyn was localized abundantly in the ectoplasmic specialization of the Sertoli cells, and Fyn was colocalized with actin filaments in the specializations.
3. Western blot analyses
Triton-soluble and insoluble fractions were prepared from the mouse testis, and the samples were applied for SDS-PAGE, transferred to the membrane, and immunostained with anti-Fyn antibody. The results indicated that Fyn protein was detected mostly in the Triton-insoluble, cytoskeletal fraction. These data strougly suggest that Fyn kinase is associated with cytoskeletal proteins. Using anti-phosphotyrosine antibody, a tyrosine-phosphorylated protein at 80 kDa(p80) was abundantly detedted in the Triton -insoluble cytoskeletal fractions of the wild-type testis. In contrast, p80 was significantly reduced in the cytoskeletal fraction of the fyn -/- testes. This finding indicated that p80 might be the target of Fyn kinase.
4. Abnormalities in the fyn -/- testis during postnatal development
Adultfyn -/- male mice are fertile. However, we found that the weight of fyn -/- immature testes at the age of 3 to 4 weeks significantly decreased compared to that of wild-type testes. The diameter of the seminiferous tubules were also decreased, and necrotic germ cells were frequently found in the fyn -/- testes. Furthermore, uncommon ultrastructures of ectoplasmic specializations were also detected in the fyn -/- testes. These findings showed that Fyn Kinase is involved in the survival and/or differentiation of the germ cells at the specific stages via Sertoli cells.
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Report
(3 results)
Research Products
(8 results)
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[Publications] Maekawa M, Suzuki-Toyota F, Toyama Y, Kadomatsu K, Hagihara M, Kuno N, Muramatsu T, Dohmae K, Yuasa S: "Stage-specific localization of basigin, a member of the immunoglobulin superfamily, during mouse spermatogenesis."Arch. Histol. Cytol.. 61. 405-415 (1998)
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