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Molecular cloning of a sperm fucosidase cDNA of Halocynthia roretzi

Research Project

Project/Area Number 06839014
Research Category

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

Allocation TypeSingle-year Grants
Research Field 海洋生物学
Research InstitutionTOKYO INSTITUTE OF TECHNOLOGY

Principal Investigator

MATSUMOTO Midori  Tokyo Institute of Technology, assistant Professor, 生命理工学部, 助手 (00211574)

Co-Investigator(Kenkyū-buntansha) HOSHI Motonori  Tokyo Institute of Technology, Profeccer, 生命理工学部, 教授 (20012411)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordsfertilization / fucosidase / 精子 / 卵精子結合
Research Abstract

Specific binding of spermatozoa to the vitelline coat is a necessary step for fertilization in various animals. In two ascidian species, Halocynthia roretzi and Ciona intestinalis, L-fucosyl residues of acidic saccharide chains in glycoproteins appear to beessential for the sperm-receptor activity of the vitelline coat. Sperm binding was blocked by specific inhibitors against fucosidase, and the fucosylamine affinity column adsorbed the sperm extract appreciably in normal sea water. It is thus suggested that sperm fucosidase play an important role in sperm binding to the vitelline coat.
In the present study, we isolated the cDNA (HrFuc) encoding Halocynthia roretzi sperm alpha-L-fucosidase by RT-PCR method using the conservative region of alpha-L-fucosidase. The nucleotide and fucosidase were compared with the human, rat and Discoidium fucosidase sequences. The polyclonal antibody was prepared against a lacZ-HrFuc fusion protein made in Escherichia coli. Western analysis showed that this antibody crossed to 33 kDa sperm proteins. And this antibody inhibited ferterization in Halocynthia roretzi. Then, the cloned cDNA seems to be closedly corelated with the sperm-vitelline coat binding.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] S.Kurata et al.: "Transcriptional control of the heme oxygenase gene in mouse M1 cells during their TPA-induced differentiation into macrophage." J.Cellular Biochemistry. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Jimbo et al.: "Protoplast Fusion Method for Efficient Transformation of Mouse Spermatogenic Cells" J.Biochem.117. 321-323 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] N.Akita: "Hemocytes Release Phenoloxidase upon Contact Reaction,an Allogeneic Interaction,in the Ascidian Halocynthia roretzi" Cell Struc.Func.20. 81-87 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Hoshi: "Allogeneic recognitrion in ascidian fertilization In ^*Sixth Lake Shirakaba Placenta Conference 1994 Biology of Pregnancy^*" Keiseisha,Tokyo, (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Hoshi, S.Takizawa and N.Hirohashi: "Glycosidases, proteases and ascidian fertilization" Seminars in DEVELOPMENTAL BIOLOGY. 5. 2001-208 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Hoshi: "Allogeneic recognition in ascidian fertilization" In ^*Sixth Lake Shirakaba Placenta Conference 1994 Biology of Pregnancy^* ed.T.Nakayama and T.Makino, pp.60-65, Keiseisha, Tokyo. (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Jimbo, M.Matsumoto, S.Kurata and M.Hoshi: "Protoplast Fusion Method for Efficient Transformation of Mouse Spermatogenic Cells" J.Biochem.117. 321-323 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] S.Kurata, M.Matsumoto: "Transcriptional control of the heme oxygenase gene in mouse M1 cells during their TPA-induced differentiation into macrophage." J.Cellular Biochemistry. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] S.Kurata et al.: "Transcriptional control of the heme oxygenase gene in mouse M1 cells during their TPA-induced differentiation into macrophage." J.Cellular Biochemistry. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Jimbo et al.: "Protoplast Fusion Method for Efficient Transformation of Mouse Spermatogenic Cells" J.Biochem.117. 321-323 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] N.Akito: "Henocytes Release Phenoloxidase Upon Contact Reaction,an Allogeneic Interaction,in the Ascidian Halocynt in the Ascidian Halocynt in roretzi" Cell struc.Func.20. 81-87 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Hoshi: "Allogeneic recognition in ascidian fertilization In Sixth Lake Shirakaba Placenta Conference 1994 Biology of Pregnancy" Keiseisha,Tokyo, (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Mitsuru Jimbo: "Protoplast Fusion Method for Efficient Transfection of Mouse Spermatogenic Cells." Journal of Biochemistry. 117. 321-323 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Motonori Hoshi: "Eggs-jelly signal molecules for triggering the acrosome reaction in starfish spermatozoa." International Journal Developmental Biology. 38. 167-174 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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