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1993 Fiscal Year Final Research Report Summary

M/lecular regulation of germ cell formation

Research Project

Project/Area Number 02102010
Research Category

Grant-in-Aid for Specially Promoted Research

Allocation TypeSingle-year Grants
Research InstitutionOkazaki National Research Institutions

Principal Investigator

NAGAHAMA Yoshitaka  Okazaki National Research Institutions, National Institute for Basic Biology, Professor, 基礎生物学研究所, 教授 (50113428)

Co-Investigator(Kenkyū-buntansha) KOBAYASHI Tohru  Okazaki National Research Institutions, National Institute for Basic Biology, As, 基礎生物学研究所, 助手 (30221972)
TANAKA Minoru  Okazaki National Research Institutions, National Institute for Basic Biology, As, 基礎生物学研究所, 助手 (80202175)
YAMASHITA Masakane  Okazaki National Research Institutions, National Institute for Basic Biology, As, 基礎生物学研究所(平成5年4月より北海道大学・理学部・助教, 助手 (30202378)
YOSHIKUNI Michiyasu  Okazaki National Research Institutions, National Institute for Basic Biology, As, 基礎生物学研究所, 助手 (50210662)
Project Period (FY) 1990 – 1993
KeywordsFish / Germ cells / Hormone / Meiosis / Gonads / Cell cycle / Maturation-promoting factor (MPF) / Cell culture
Research Abstract

The molecular mechanism of germ cell formation, in particular spermatogenesis and oocyte maturation, has been investigated, using fish as a primary study model.
1) The cDNAs encoding 5 steroidogenic enzymes responsible for 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17a, 20b-DP, a common maturation-inducing hormone in fish) biosynthesis have been isolated and sequenced. We have shown that gonadotropin-induced 20beta-HSD mRNA expression is a prerequisite for 17a, 20b-DP production by granulosa cells. 2) We characterized specific binding of [^3H]17a, 20b-DP to plasma membranes from defolliculated oocytes of rainbow trout. A gonadotropin-induced increase in 17a, 20b-DP receptor concentrations is responsible for the development of oocyte maturational competence. Carp maturation-promoting factor (MPF) (cdc2 kinase and cyclin B) was purified and characterized. 17a, 20b-DP induces oocytes to synthesize cyclin B, which in turn activates preexisting cdc2 kinase through threonine phosphorylation (161) of cdc2 kinase by a threonine kinase (p40^<M015>). We discovered that cyclin degradation is triggered by limited proteolysis of cyclin B (Lys57) by active proteasome. 3) An organ culture system has been used to demonstrate that in the eel 11-ketotestosterone can induce all stages of spermatogenesis in vitro within 21 days. Our findings suggest that gonadotropin stimulates the Leydig cells to produce 11-ketotestosterone, which in turn activates the Sertoli cells to produce activin B.Activin B then acts on spermatogonia to induce mitosis leading to the formation of spermatocytes.

  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Miura,T.et al.: "Hormonal induction of all stages of spermatogenesis in vitro in the male Japanese eel (Anguilla japonica)" Proc.Natl.Acad.Sci.USA. 88. 5774-5778 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Choi,T.et al.: "Activation of p34cdc2 protein kinase activity in meiotic and mitotic cell cycles in mouse oocytes and embryos" Development. 113. 789-795 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamashita,M.et al.: "Purification and characterization of maturation-promoting factor in fhish" Developmental Biology. 249. 8-15 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tanaka,M.et al.: "Pig testicular 20β-hydroxysteroid dehydrogenase exhibits carbonyl reductase-like structure and activity:cDNA cloning of pig testicular 20β-hydroxysteroid dehydrogenase" Journal of Biological Chemistry. 267. 13451-13455 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirai,T.et al.: "Cyclin B in fish oocytes:its cDNA and amino acid sequences,appearance during maturation of goldfish oocytes." Molecular Reproductive and Development. 33. 131-140 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Katsu,Y.et al.: "Behaviour of the components of maturation-promoting factor,cdc2 kinase and cyclin B,during oocyte maturation of goldfish" Developmental Biology. 160. 99-107 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miura, T.et al.: "Hormonal induction of all stages of spermatogenesis in vitro male Japanese eel (Anguilla Japonica)." Proc. Natl. Acad. Sci. USA. 88. 5664-5778 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Choi, T.et al.: "Activation of p34^<cdc2> protein kinase activity in meiotic and mitotic cell cycles in mouse oocytes and embryos." Development. 113. 789-795 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamashita, M.et al.: "Purification and characterization of maturation-promoting factor in fish." Develop. Biol.249. 8-15 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tanaka, M.et al.: "Pig testicular 20b-hydroxysteroid dehydroganase exhibits carbonyl reductase-like structure and activity : cDNA cloning of pig testicular 20b-hydroxysteroid dehydroganase." J.Biol. Chem.267. 13451-13455 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirai, T.et al.: "Cyclin B in fish oocytes : its cDNA and amino acid sequences, appearance during maturation of goldfish oocytes." Mol. Reprod. Develop.33. 131-140 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Katsu, Y.et al.: "Behavior of the components of maturation-promoting factor, cdc2 kinase and cyclin B, during oocyte maturation of goldfish." Develop. Biol.160. 99-107 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshikuni, M.et al.: "Specific bindings of [^3H]17a, 20b-dihydroxy-4-pregnen-3-one to oocyte cortices of rainbow trout (Oncorhynchus mykiss)." Fish Physiol. Biochem.11. 15-24.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kajiura, H.et al.: "Isolation and characterization of goldfish cdc2 kinase, a catalytic component of maturation-promoting" factor. Develop. Growth Differ.35. 647-654.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fukada, S.et al.: "Steroidogenesis in the ovarian follicle of medaka (Oryzias latipes, a daily spawner) during oocyte maturation." Develop. Growth Differ.36. 81-88.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshikuni, M.et al.: "Involvement of an inhibitory G-protein in the signal transduction pathway of maturation-inducing hormone (17a, 20b-dihydroxy-4-pregnen-3-one) action in rainbow trout (Oncorhynchus rhodurus) oocytes." Develop. Biol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mita, M.et al.: "G-proteins and adenylate cyclase in ovarian granulosa cells of amago salmon (Oncorhynchus rhodurus)." Mol. Cell. Endocrinol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1995-03-27  

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