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Molecular approach to mammalian fertilization and its application to gene transfer into fertilized egg

Research Project

Project/Area Number 02454523
Research Category

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

Allocation TypeSingle-year Grants
Research Field Laboratory animal science
Research InstitutionUniversity of Tokyo

Principal Investigator

MORI Tsuneatsu  Univ.of Tokyo Inst.of Med.Sci. Associate Prof., 医科学研究所, 助教授 (40012760)

Co-Investigator(Kenkyū-buntansha) AMANO Takashi  Tokyo Univ.of Agriculture Associate Prof.(1990-1992), 畜産学科, 助教授 (90078147)
NOSE Kiyoshi  Univ.of Tokyo Inst.o Med.Sci. Associate Prof.(1990-1991), 医科学研究所, 助教授 (70012747)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1992: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1991: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsZona pellucida / Sugar chain of Zona pellucida / vitelline membrane / CD4 of vitelline membrane / P56^<lck> of vitelline membrane / MHC class II molecule of sperm / DNA binding molecule of sperm / Signal transduction in fertilization / 透明帯中性糖鎖 / 透明帯酸性糖鎖 / 透明帯結合蛋白 / MHCクラスII分子 / CD_4分子 / P56^<lck> キナーゼ / 受精の情報伝達 / 卵透明帯 / 糖鎖 / 精子 / 卵子 / CD_4〓分子 / P56lck / 哺乳類 / 中性糖 / クラスIIMHC抗原 / 外来遺伝子結合性 / 卵黄膜 / L_3T_4原分子
Research Abstract

By means of monoclonal technology, we obtained six hybridoma cell lines producing mAbs to the porcine zona pellucida (ZP). The epitopes recognized by these mAbs were chemically characterized. Four of them recognized a porcine ZP-specific primary structure of protein and two of them recognized a steric structure of the protein similar in both porcine and human ZPs.
Furthermore, the biochemical analysis of carbohydrate structure of porcine ZP revealed that considerable quantities of N-linked, neutral oligosaccharides in porcine ZP were terminated with N-acetylglucosamine residues, suggesting the possible roles of these sugar chains as the acceptors for the galactosyltransferase on sperm.
Following the studies of sperm acceptor on ZP, we found the expression of MHC class II molecule on the posterior region of sperm and its possible roles of foreign DNA binding to sperm.
Corresponding to the MHC class II molecule on sperm, CD4 molecule associated with p56^<lck> was definitely observed on the vitelline membrane of egg. Besides, the adherence between the posterior region of sperm head and the vitellus of egg was mediated by MHC class II molecule of sperm and CD4 molecule of egg and their signal into egg activation was transduced through p56^<lck> at the fusion step of murine fertilization.
Actually, RNA transcript in murine eggs corresponding to parts of the CD4 gene and lck gene in thymocytes were obtained by using RT/PCR methods followed by the sequencing of PCR products. The deduced sequences potentially encode the aminoacid sequence of transmembrane to cytoplasmic domain of CD4 and that of N-terminal domain of p56^<lck> respectively. These results demonstrated that the signal transducing complex similar to that in immune T cells are expressed at the transcriptional levels in murine eggs.

Report

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

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Mori.T et.al.: "Expression of CD_4-like structure on murine egg vitelliue membrane and its signal transductive roles through P56^<lck> in fertilization" Am.J. Reprod Immunol. 26. 97-103 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mori.T.et.al.: "Expression of the signal transducing regions of CD_4-like and lck genes in murine egg" Biochem.Biophys.Res.Commun. 182. 527-533 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mori.T.et.al.: "Studies of mammalian implantation in vitro (I)-Production of laminin from human decidual cell line and its effects on murinetrophoblastic outgrow" J.Reorod.Dev.38. 285-292 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mori.T.et.al.: "Studies of Mammalian implantation in vitro (II)-Effects of cytokiues from mixedlymphocyte reaction on Proliteration of murine tropuoblast cells and establishment of ifs cell liue" J.Reorocl.Dev. 38. 293-301 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kamada.M.etal.: "Hetes plile autigens in serous cystadenocancinma of the wuman ovary" Asia-Oceania J.Obset.Gynaecal. 18. 387-395 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 森 庸厚他: "生殖・腫瘍免疫と糖鎖" 蚕白質・核酸・酸素(増刊号). 37. 1772-1782 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 森 庸厚.他: "生殖系列-親から子への生命の流れ 第七回「大学と科学」公開シンポジウム構演収録集" 「大学と科学」公開シンポジウム組織委員会(クバプロ), (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mori.T et.al.: "Immunological studies on the mammalian fertilization and implantation in Biology of germ lines-in animals amd men" Japan sci.soc.Press Tokyo, (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mori,E.et.Al.: "Neutral oligosaccharide structures linked to asparagines of porcine zona pellucida glycoproteins" Biochemistry. 30. 2078-2087 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.et.al.: "Expression of CD4ーlike structure on murine egg vitelline membrane and its signal transductive roles through p56lck in fertilization" Am.J.Reprod.Immunol.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.et.al.: "Expression of the signal transducing region of CD4ーlike and lck genes in murine egg" Biochem.Biophys.Res.Commun.182. 527-533 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.: "細胞性免疫" 臨床婦人科産科. 46. 142-144 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.et.al.: "生殖・腫瘍免疫と糖鎖" 蛋白質 核酸 酵素. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.: "Immunobiological studies on the mammalian fertilization and implantation.In “Biology of Germ Linesーin Animals and Man"" Jap.Sci.Soc.Press, (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mori,T.,et al.: "Expression of class II major histocompatibillity complex antigen on mouse sperm and its roles in fertilization" Am.J.Reprod.Immunol.24. 9-14 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Gou,M.W.,et al.: "Binding of foreign DNA to mouse sperm mediated by its MHC class II structure" Am.J.Reprod.Immunol.24. 120-126 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Mori,E.,et al.: "Neutral oligosaccharide structures linked to asparagines of porcine zona pellucida gycoproteins" Biochemistry. (1991)

    • Related Report
      1990 Annual Research Report

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

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