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分子生物学的アプローチによる哺乳類卵透明帯の構造解析

Research Project

Project/Area Number 05671350
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionYamagata University

Principal Investigator

荒木 慶彦  山形大学, 医学部, 助手 (70250933)

Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywords透明帯 / ラット / マウス / 精子・卵相互作用 / 受精
Research Abstract

哺乳動物の卵透明帯(ZP)は、種特異的な精子・卵相互作用に関与する重要な構造物であり、その分子構造の解明は、受精の分子機構解明のために必須である。本研究では今まで実験動物として最もポピュラーなラットとマウスのZPの構造を比較検討する事でその種特異的精子・卵相互作用の分子機構の解明を試みてきたが、現在まで以下の結果を得た。1)マウスZP2とZP3上にはN結合型poly-N-acetyllactosaminyl glycanとO結合型trisaccharideが存在し特に従来その精子・卵結合に於ての重要性が提唱されていたZP3上のO結合型糖鎖はWassarmanらの報告よりも、その含有量は遙に少なく、しかもその構造はGlcNacβ→Galβ1,3GalNAcolであった。この結果は従来定説化していたマウスZP3のO結合型糖鎖上のαGal残基が精子へのリガンドとする説を真正面から疑問視する結果となり、国際的にも注目されている研究結果である。2)マウスZP2及びZP3cDNA(米国NIH Dr.J.Deanより供与)をprobeとしてラット卵巣cDNAライブラリーをスクリーニグした。これらのprobeを用いたラット卵巣RNAに対するNorthern blottingではそれぞれ2.2kb,1.3kbのメッセージを認めたが、スクリーニングの結果、陽性クローンの最長のものはZP2は2.2kbp,ZP3は1.05kbpであった。現在、これらのシークエンスの全貌を明らかにする為に、欠損部のRapid Amplification of cDNA endを含めその最終段階に入っている。

Report

(2 results)
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] NagDas S.K.et.al.: "O-linked trisaccharide and N-linked poly-N-acetyllactosaminyl glycans are present on mouse ZP2 and ZP3." Biology of Reproduction. 51. 262-272 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Tulsiani D.R.P. et.al.: "Purification and characterization of two forms of β-D-galactosidase from rat epididymal luminal fluid:Evidence for their role in the modification of sperm plasma membrane glycoprotein(s)." Biochemical Journal. in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Suzuki K.et.al.: "Molecular characterization of a hamster oviduct specific glycoprotein." Biology of Reproduction. in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Nohara M.et.al.: "In vitro fertilization in mastomys(Praomys coucha)" Biology of Reproduction. in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Sendai Y.et.al.: "Molecular cloning and characterization of a mouse oviduct specific glycoprotein." Biology of Reproduction. in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Komiya H.et.al.: "Identification of hamster oviduct-specific glycoprotein messenger ribonucleic acid by in situhybridization" Biology of Reproduction. in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Araki Y: "Qualitative characterization of oligosaccharide chains present on the rat zona pollucida glycoconjugates" Biology of Reproduction. 46. 912-919 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Araki Y: "Isolation and characterization of a 25kilodalton protein from mouse testes;Sequence homology with a phospholipid binding protein" Biology of Reproduction. 47. 832-843 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Vierula ME: "Immunolocatization of a 25kilodalton protein in mouse testis and epididymis" Biology of Reproduction. 47. 844-856 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 荒木慶彦: "哺乳類精子・卵相互作用:配偶子の成熟・結合機構と糖鎖" 蛋白質・核酸・酵素. 38. 2347-2357 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yano K: "Boundary of autoinhibitory region of smooth muslce myosin light chain kinase" Biochemistry. 32. 12054-12061 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kamisoyama H: "Mutagenesis of the phosphorylation site(Serine 19)of smooth muscle myosin regulatory light chain and its ettects on the properties of myosin" Biochemistry. 33. 840-847 (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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