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Analyses of the localization of germ plasm and purification of "germ cell determinants" in amphibia.

Research Project

Project/Area Number 60540452
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 動物発生・生理学
Research InstitutionHokkaido University

Principal Investigator

WAKAHARA Masami  Zool. Inst., Fac. Sci., Hokkaido University, 理学部, 助教授 (00001868)

Project Period (FY) 1985 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1986: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1985: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsXenopus laevis / Primordial germ cells (PGCs) / Germ plasm / FDA / Monoclonal antibody / "生殖細胞決定因子" / 始原生殖細胞 / 生殖質 / 螢光標識デキストラン-アミン(FDA)
Research Abstract

1. Correlation of the number of primordial germ cells (PGCs) at stage 47 with the amount of germ plasm at the 8-cell stage was analysed using two different laboratory-rased colonies of Xenopus laevis, HD and G groups. The average number of PGCs in J group tadpoles was significantly larger than that in HD group tadpoles. The amount of germ plasm was also demonstrated to be larger than in HD group. The amount of germ plasm was related positively to the number of PGCs. Embryos which contained larger amounts of germ plasm developed larger numbers of PGCs at stage 47. These findings suggest that the number of PGCs is regulated by at least two different mechanisms; first, the number of PGCs is primarily specified by the intrinsic amount of germ plasm in the fertilized egg. Second, it is regulated by an unknown mechanism which controls the totoal number of cells of whole embryos, such as the nucleocytoplasmic ratio.
2. A reliable biological assay system for "germ cell determinants" was newly developed. When 90 -rotated eggs were developed to 2 cell stage, fluorescence-dextran-amine (FDA), a reliable cell tracer, was injected into one blastomer, originally animal- or vegetal-blastomere. As a control, FDA was injected into one blastomere of normally oriented egg. After reaching stage 47, tadpoles were examined for the fluorescence. In the control tadpoles, about half of the PCSs had a strong fluorescence from FDA. On the other hand, no fluorescence was detected in the PGCs of the tadpoles which had been injected with FDA into the original animal-hemispere.
3. Monoclonal antibodies is going to be raised against cytoplasmic fraction including the germ plasm: 20.000xg precipitate cytoplasmic fraction from the gegetal hemispere of the fertilized eggs were injected into Balb/C mice, and the splenocytes were fused tomyeloma cells to get potency hybrydomas.

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] AKITA,Y.AND WAKAHARA,M.: J.Embryol.exp.Morph.90. 251-265 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: Devel.Growth,Differ.28. 543-554 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] KUNIEDA,M.AND WAKAHRA,M.: Zool.Sci.4. 489-496 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: Proc.First NASA-Japan Space Biology Workshop. 1. 99-106 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 若原正己: 数理科学.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: Primers in Developmental Biolgy. 4. (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 文部省特定研究「咀嚼システムの基礎研究」総括班編: "咀嚼システム入門" 風人社, PP.11-23 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] AKITA, Y. and M. WAKAHARA: "Cytoplasmic analyses of factors which determine the number of primordial germ cells (PGCs) in Xenopus laevis." J. Embryol. exp. Morph.90. 251-265 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: "Modification of dorsal-ventral polarity in Xenopus laevis embryos following withdrawal of egg contents before first cleavage." Devel. Growth, Differ.28. 543-554 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] KUNIEDA, M. and M. WAKAHAWA: "Twin formation in Xenopus laevis eggs centrifuged before first cleavage." Zool. Sci.4. 489-496 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: "Possible plans for space biology experiments using amphibian eggs." Pro. First NASA-Japan Space Biol. Workshop. 1. 99-106 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] WAKAHARA,M.: "Cytoplasmic localization and organization of germ cell determinants." Primers in Devel. Biol.4. (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] AKITA,Y.;M.WAKAHARA: J.Embryol.exp.Morph.90. 251-265 (1985)

    • Related Report
      1986 Annual Research Report
  • [Publications] WAKAHARA,M.: Develop.Growth,Differ.28. 543-554 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] KUNIEDA,M.;M.WAKAHARA: Zool.Sci.4. (1987)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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