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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
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥175,000,000 (Direct Cost: ¥175,000,000)
Fiscal Year 1993: ¥33,000,000 (Direct Cost: ¥33,000,000)
Fiscal Year 1992: ¥33,000,000 (Direct Cost: ¥33,000,000)
Fiscal Year 1991: ¥50,000,000 (Direct Cost: ¥50,000,000)
Fiscal Year 1990: ¥59,000,000 (Direct Cost: ¥59,000,000)
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.

Report

(5 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (42 results)

All Other

All Publications (42 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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [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
    • Related Report
      1993 Final Research Report Summary
  • [Publications] N.Shibata,M.Yoshikuni &Y.Nagahama: "Vitellogenin incorporation intooocytes of rainbow trout,Oncorhynchus mykiss,in vitro effect of hormones on dunuded cocytes." Develop.Growth Differ. 35. 115-121 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Nakamura,J.L.Specker & Y.Nagahama: "Ultrastructural analysis of formation of a functional follicle during early vitellogenesis in the tilapia,Orepchromis niloticus." Cell Tissue Res.272. 33-39 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Takahashi,M.Tanaka,N.Sakai,S.Adachi,W.L.Miller & Y.Nagahama: "Rainbow trout ovarian choesterol side‐chain cleavage cytochrome p450(p450scc):cDNA cloning and mRNA expresion during oogenesis." FEBS Lett.319. 45-48 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Kobayashi,N.Sakai,S.Adachi,H.Iwasawa,K.Asahina & Y.Nagahama: "17alpha,20beta‐Dihydroxy‐4‐pregen‐3‐one is the naturally occurring spermiationinducing hormone in the testis of a frog,Rana nigromaculata." Endocrinology. 133. 321-327 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Iwao,N.Sakamoto,K.Takahara,M.Yamashita & Y.Nagahama: "The egg nucleus regulates the behabior of sperm nuclei as well as cycling of MPF in physiologically polyspermic newt eggs." Develop.Biol.160. 15-27 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Katsu,M.Yamashita,H.Kajiura & Y.Nagahama: "Behavior of the components of maturation‐promoting factor,cdc2 kinase and cyclin B during oocyte maturation of goldfish." Develop.Biol.160. 99-107 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yamashita,M.,Fukada,Yoshikuni,M.et al.: "Purification and characterization of maturation-promoting factor in fish" Develop.Biol.149. 8-15 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Tanaka,M.,Telecky,T.M.,Fukada,S.: "Cloning and sequence analysis of the cDNA encoding P-450 aromatase (450arom) from a rainbow trout --." J.Mol.Endocrinol.8. 53-61 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Sakia,N.et al.: "Rainbow trout cytochrome P-450c17 (17α-hydroxylase/17,20-lyase):cDNA cloning,enzymatic properties and temporal pattern of ovarian P-450c17 mRNA----." FEBS Letter. 301. 60-64 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Yamashita,M.: "M-phase-specific histone Hl Kinase in fish oocytes:its purification,components and biochemical properties." Eur.J.Biochem.205. 537-543 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hirai,T.et al.: "Isolation and characterization of goldfish cdk2,a cognate variant of the cell cycle regulator cdc2." Develop.Biol.152. 113-120 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Tanaka,K.et al.: "Pig testicular 20β-hydroxysteroid dehydrogenase exhibits carbonyl reductase-like structure and activity:cDNA cloning of pig testicular 20β-hydroxysteroid - -." J.Biol.Chem.267. 13451-13455 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Miura,K.Yamauchi,H.Takahashi,& Y.Nagahama: "Hormonal induction of all stages of spermatogenesis in vitro in the male japanese eel (Anguilla japonica)" Prodeedings of the National Academy of Sciences,U.S.A.88. 5774-5778 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] A.Yamaguchi,M.Yamashita,M.Yoshikuni,Y.Hotta,P.Nures,& Y.Nagahama: "Involvement in meiotic prophase of Hl histone kinase and p34cdc2 homologues in lily (Lilium longiflorum) microsporocytes" Development,Growth and Differentiation. 33. 625-632 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Yamashita,M.Yoshikuni,T.Hirai,S.Fukada,& Y.Nagahama: "A monoclonal antibody against the OSTAIR sequence of p34cdc2,catalytic subunit of maturation-promoting factor and key regulator of the cell cycle" Development,Grwoth and Differentiation. 33. 617-624 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Yamashita,S.Fukada,M.Yoshikuni,P.Bulet,T.Hirai,- - & Y.Nagahama: "Purifaction and characterization of maturation-promoting factor in fish" Developmental Biology. 149. 8-15 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Tanaka,T.M.Telecky,S.Fukada,S.Adachi,S.Chen,& Y.Nagahama: "Cloning and sequence analysis of the cDNA encoding P-450 aromatase (P450arom) from a rainbow trout (Oncorhynchus rhodurus) ovary:ralationship between the amount of - in the ovary" Journal of Molecular Endocrinology. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Sakai,M.Tanaka,S.Adachi,W.L.Miller,& Y.Nagahama: "Rainbow trout cytochrome P450c17(17αーhydroxylase/17,20ーlyase):cDNA cloning,enzymatic properties and temporal pattern of ovarian P450c17 mRNA expression during oogenesis" FEBS Letters. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 長浜 嘉孝(分担:生殖の項): "魚類生理学(板沢 靖男・羽生 功 編)" 恒星社厚生閣, 243-286 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Nagahama,A.Matsuhisa,T.Iwamatsu,N.Sakai,& S.Fikada: "A mechanism for the action of pregnant mare serum gonadotropin on aromatase activity in the ovarian follicle of the medaka,Oryzias latipes" Journal of Experimental Zoology. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] T.Miura,K.Yamauchi,H.Takahashi,& Y.Nagahama: "Hormonal induction of all stages of spermatogenesis in vitro in the male Japanese eel (Anguilla japonica)" Proceedings of the National Academy of Sciences,U.S.A.(1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] M.Mori,M.Yamashita,S.Fukada,M.Yoshikuni & Y.Nagahama: "Maturationーpromoting factor and p34cdc2 kinase during oocyte maturation of the Japanese quail" Developmental Biology. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] M.Yamashita,S.Fukada,M.Yoshikuni,T.Hirai,ーー& Y.Nagahama: "Purification and characterization of maturationーpromoting factor in fish" Developmental Biology. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] S.Adachi & Y.Nagahama: "Involvement of pH in the enhancement of 20βーhydroxysteroid dehydrogenase activity in isolated granulosa cell layers from ovarian follicles of salmonid fishes" Biomedical Research. 11. 327-335 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Biomedical Research: "Identification of aromatizable androgens secreted by isolated thecal cell layers from vitellogenic ovarian follicles of a teleost,amago salmon (Oncorhynchus rhodurus)" Biomedical Research. 11. 359-363 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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