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Abnormal morphology of the testes of the fyn-deficient mice and analysis of the function of Fyn tyrosine kinase

Research Project

Project/Area Number 12670007
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionCHIBA UNIVERSITY

Principal Investigator

MAEKAWA Mamiko  Chiba University, Graduate School of Medicine, Assistant, 大学院・医学研究院, 助手 (20181571)

Co-Investigator(Kenkyū-buntansha) TOYAMA Yoshiro  Chiba University, Graduate School of Medicine, Lecturer, 大学院・医学研究院, 講師 (70009637)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsFyn / tyrosine kinase / testis / Sertoli cell / spermatogenesis / knockout mouse
Research Abstract

1. Significant reduction of the testis weight and degenerated germ cells were found at the age of 3 and 4 weeks of fyn-deficient mice. Electron microscopy revealed that fyn-deficient testis showed ultrastructural abnormality in specialized junctional structures of the Sertoli cells, termed the ectoplasmic specializations (ES). Small unusual vesicles were found in the actin filament layer of the ES. Immunohistochemical studies demonstrated that Fyn was concentrated together with actin filaments at the ES. At these sites high level of phosphotyrosine was also immunostained in the wild type testes, while phosphotyrosine immunoreactivity was decreased in the fyn -deficient testes. These findings suggest that Fyn kinase functions at the ectoplasmic specializations of the Sertoli cells in the testis.
2. Immunoblot analyzes revealed that Fyn was distributed mostly in the Triton X-100 insoluble cytoskeletal fraction prepared from the wild-type mouse testis, suggesting that Fyn may be associated with cytoskeletal proteins such as actin filaments. A tyrosine phophorylated protein at around 80 kDa (P80) was prominent in the Triton-insoluble fraction from wild-type testis. However, P80 was specifically decreased in the fyn-deficient testes. This finding indicated that P80 might be the target of Fyn kinase, and that P80 may play a role in a Fyn-mediated signal transduction in the mouse testis.
3. We found that a mouse Sertoli cell line, TM4, expressed Fyn protein. Tyrosine-phosphorylated proteins were examined in the TM4 cell extracts by two-dimensional immunoblot analysis. When the cells was treated with environmental endocrine disrupters, such as diethylstilbestrol, the level of tyrosine phosphorylation of one protein at approximately 38kDa (P38) was conspicuously increased compared to the control cells. It is possible that Fyn tyrosine kinase may be involved in the phosphorylation of the P38.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] M.Maekawa, et al.: "Fyn tyrosine kinase in sertoli cells is involved in mouse spermatogenesis"Biology of Reproduction. 66. 211-221 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y.Toyama, et al.: "Neoratally administered diethylstilbestrol retards the development of the blood-testis barrier in the rat"Journal of Andrology. 22. 413-423 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] J.Yuasa, et al.: "Specific localization of the basigin protein in human testes from normal adults, normal juveniles, and patients with azoospermia"Andrologia. 33. 293-299 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toyama Y, Ohkawa M, Oku R, Maekawa M, Yuasa S: "Neonatally administrated diethylstilbestrol (DES) retards the development of the blood-testis barrier in the rat."J. Androl. 22. 413-423 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yuasa J,Toyama Y, Miyauchi T, Maekawa M, Yuasa S, Ito H: "Specific localization of the basigin protein in human testes from normal adults, normal juveniles, and patients with azoospermia"Andrologia. 33. 293-299 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Maekawa M,Toyama Y, Yasuda M, Yagi T, Yuasa S: "Fyn tyrosine kinase in Sertoli cells is involved in mouse spermatogenesis."Biol Reprod. 66. 211-221 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Maekawa et al.: "Fyn tyrosine kinase in Sertoli cells is involved in mouse spermatogenesis"Biology of Reproduction. 66. 211-221 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y. Toyama et al.: "Neonatally administered diethylstilbestrol retards the development of the blood-testis barrier in the rat"Journal of Andrology. 22. 413-423 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] J. Yuasa et al.: "Specific localization of the basigin protein in human testes from normal adults,normal juveniles,and patients with azoospermia"Andrologia. 33. 293-299 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Toyama et al.: "Neonatally Administered Diethylstilbestrol Retards the Development of the Blood Testis Barrier in the Ret"J.Andrology. (in press). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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