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Study on regulatory factors involved in fertilization and development

Research Project

Project/Area Number 03454490
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionHokkaido University

Principal Investigator

YOKOSAWA Hideyoshi  Hokkaido University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (90012765)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1991: ¥5,100,000 (Direct Cost: ¥5,100,000)
KeywordsFertilization / Development / Proteasome / GTP-binding protein / アネキシン / グルコシダーゼ / プロテア-ゼ / プロテアソ-ム / GTPー結合蛋白質 / リポコルチン / グルコシダ-ゼ
Research Abstract

Fertilization is initiated by sperm-egg interaction, fusion of egg with a single sperm activates the egg, and embryonic development ensues. The progression of fertilization and development is regulated by many regulatory proteins. We have investigated the physiological roles of proteasome (high molecular weight protease complex) and small GTP-binding protein in the process including (1) sperm penetration through egg vitelline coat, (2) elevation of egg vitelline coat as a result of egg activation, and (3) cleavage of fertilized eggs. 1. We isolated two kinds of chymotrypsin-like enzymes from ascidian sperm. One is characterized as a typical 20 S proteasome, while the other is a larger protease comples that is cross-reacted with anti-proteasome antibody but shows different inhibitor susceptibility from that of 20 S proteasome. These proteasomes were found to be released from sperm upon the sperm reaction and the released proteasomes had the activity to digest the egg vitelline coat. 2. Microinjection of botulinum exoenzyme C3 that ADP-ribosylates small GTP-binding proteins induced elevation of the vitelline coat of ascidian eggs, whereas that of monoclonal antibody that inhibits C3-catalyzed ADP-ribosylation inhibited the elevation induced by insemination. These results suggest the involvement of Rho family of small GTP-binding proteins in the activation of ascidian eggs. 3. Immunocytochemistry using monoclonal antibody against ascidian egg proteasome shows that proteasome undergoes a cell cycle- dependent change during cleavage of ascidian fertilized eggs. Proteasome activity also was activated during cleavage. Theses results suggest that proteasome participates in the progression of cell cleavage cycle during ascidian embryonic development.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Yoshiko Saitoh: "Comparative studies on proteasomes (multicatalytic proteasomes) isolated from spermatozoa and eggs of sea urchins." Comp.Biochem.Physiol.99B. 71-76 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hiroyuki Kawahara: "Cell cycle-dependent change of proteasome distribution during embryonic development of the acidian Halocynthia roretzi." Dev.Biol.151. 27-33 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hiroyuki Kawahara: "The 26 S proteasome is activated at two points in the ascidian cell cycle." FEBS Lett.310. 119-122 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Yoshiko Saitoh: "High molecular weight protease complexes (proteasomes) of sperm of the ascidian,Halocynthia roretzi:" Dev.Biol.in press. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 沢田 均: "受精・発生に関与するプロテアーゼの機能." 蛋白質核酸酵素. 36. 814-819 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y. Saitoh, H. Kawahara, H. Miyamatsu, and H. Yokosawa: "Comparative studies on proteasomes (multicatalytic proteinases) isolated from spermatozoa and eggs of sea urchins." Comp. Biochem. Physiol.99B. 71-76 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Kawahara and H. Yokosawa: "Cell cycle-dependent change of proteasome distribution during embryonic development of the ascidian, Halocynthia roretzi." Dev. Biol.151. 27-33 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Kawahara, H. Sawada, and H. Yokosawa: "The 26 S proteasome is activated at two points in the ascidian cell cycle." FEBS Lett.310. 119-122 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y. Saitoh, H. Sawada, and H. Yokosawa: "High molecular weight protease complexes (proteasomes) of sperm of the ascidian, Halocynthia roretzi: Isolation, characterization, and physiological roles in fertilization." Dev. Biol.(1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Sawada and H. Yokosawa: "Physiological roles of proteases in fertilization and development." Protein, Nucleic acid and Enzyme. 36. 814-819 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hiroyuki Kawahara: "The 26 S proteasome is activated at two points in the ascldian cell cycle." FEBS Lett.310. 119-122 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Yoshiko Saitoh: "High molecular weight protease complexes (Proteasomes) of sperm of the ascidian,Halocynthia roretzi." Dev.Biol.in press. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hiroyuki Kawahara: "Cell cycleーdependent change of proteasome distribution during embryonic development of the ascidian,Halocyntia roretzi." Dev.Biol.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yoshiko Saitoh: "Comparative studies on proteasomes(multicatalytic proteinases)isolated from spermatozoa and eggs of sea urchins." Comp.Biochem.Physiol.99. 71-76 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 沢田 均: "受精・発生に関与するプロテア-ゼの機能" 蛋白質核酸酵素. 36. 814-819 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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