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Analysis of inhibitory mechanism of fertilization between different species

Research Project

Project/Area Number 07456131
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Basic veterinary science/Basic zootechnical science
Research InstitutionThe University of Tokyo

Principal Investigator

HAYASHI Yoshihiro  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (90092303)

Co-Investigator(Kenkyū-buntansha) OGAWA Kenji  The University of Tokyo, The Institute of Physical and Chemical Research, Labora, 動物試験室, 研究員 (50251418)
KUROHMARU Masamichi  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Asso, 大学院・農学生命科学研究科, 助教授 (00148636)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 1996: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1995: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordsfield vole / spermatozoa / IVF / zona pellucida / hypotaurine / insemination / acrosome reaction / zona-hydrolysing enzyme
Research Abstract

First, we tried to establish the in vitro fertilization (IVF) technique of the Japanese field vole. It is improved by sperm concentration (1-2x10^7 cell/ml for preincubation and 1x10^6 cell/ml for insemination) and hypotaurine (0.1-1 mM). Preincubation of spermatozoa with 1 mM hypotaurine pripor to oocytes insemination did not have a significant effect on fertilization rate. At 96 h after insemination, most cultured embryos developed to the 2-cell stage (68-83%), but development to blastocyst was very low (0-3%) for all treatments. Althogh 43 living offspring were produced after transfer of IVF pronuclear embryos to recipients, the delivery rate of recipients was very low (21%). The litter size of pseudopregnat recipients that produced a litter was 4 to 11, and the rate of in vitro development of transferred embryos in the pregnant recipients was 41%. It would appear that hypotaurine affects IVF rather than spermatozoa preincubation and that IVF embryos can develop to normal offspring. Next, we investigated the mechanism of penetration by field vole spermatozoa of mouse and hamster zonae pellucidae. The field vole spermatozoa that can penetrate the zonae pellucidae of the mouse and hamster could not penetrate the zonae of the rat, mastomys and Mongolian gerbil. The vole spermatozoa that bound to the zona oaf field vole oocytes underwent the acrosome reaction before passing through it. In contrast, the vole spermatozoa that bound to the zonae of mouse and hamster oocytes penetrated the zonae without any sign of the acrosome reaction. The presence or absence of proteinase/hyaluronidase inhibitors in the medium did not make any difference to zona penetration by acrosome-intact vole spermatozoa. These observations suggest that field vole spermatozoa can cross mouse and hamster zonae mechanically without assistance from zona-hydrolysing enzymes.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization and embrryo development of Japanese field voles (Microtus montebelli)" Journal of Reproduction and Fertility. 104(1). 63-68 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama,Hayashi et al.: "Effect of hypotaurine on in vitro fertilization and production of term offspring from in vitro fertilized ova of Japanese field vole" Biology of Reproduction. 54(3). 625-630 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization/embryo transfer (IVF/ET) of Japanese field vole,Microtus montebelli." Advances in contraceptive delivery systems. 12(1). 69-77 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama.Hayashi et al.: "Penetration by field vole spermatozoa of mouse and hamster zonae pellucidae without acrosome reaction" Journal of Reproduction and Fertility. 107(1). 97-102 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama, T., Suto, J., Matubara, Y., Imamura, K., Fukuta, K., Toyoda, Y., Kurohmaru, M.& Hayashi, Y.: "In vitro fertilization and embryo development of Japanese field voles (Microtus montebelli)" J.Reprod.Fertil.104(1). 63-68 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama, T., Suto, J., Imamura, K., Toyoda, Y., Kurohmaru, M.& Hayashi, Y.: "Effect of hypotaurine on in vitro fertilization and production of term offspring from in vitro-fertilized ova of the Japanese field vole, Microtus montebelli" Biol.Reprod.54(3). 625-630 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama, T., Suto, J., Kurohmaru, M.& Hayashi, Y.: "In vitro fertilization/embryo transfer (IVF/ET) of Japanese field vole, Microtus montebelli" Advances in contraceptive delivery systems. 12(1). 69-77 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama, T., Ogura, A., Suto, J., Matubara, Y., Kurohmaru, M., Hayashi, Y.& Yanagimachi, R.: "Penetration by field vole spermatozoa of mouse and hamster zonae pellucidae without acrosome reaction" J.Reprod.Fertil.107(1). 97-102 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization and embryo development of Japanese field voles(Microtus montebelli)" Journal of Reproduction and Fertility. 104(1). 63-68 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "Effect of hypotaurine on in vitro fertilization and production of term offspring from in vitro fertilized ova of Japanese field vole" Biology of Reproduction. 54(3). 625-630 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization/embryo transfer(IVF/ET)of Japanese field vole,Microtus montebelli" Advances in contraceptive delivery systems. 12(1). 69-77 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "Penetration by field vole spermatozoa of mouse and hamster zonae pellucidae without acrosome reaction" Journal of Reproduction and Fertility. 107(1). 97-102 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization and embryo development of Japanese field voles (Microtus montebelli)" Journal of Reproduction and Fertility. 104. 63-68 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "Effect of hypotaurine on in vitro fertilization and production of term offspring from in vitro fertilized ova of Japanese field vole" Biology of Reproduction. (in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wakayama,Hayashi et al.: "In vitro fertilization/embryo transfer(IVF/ET) of Japanese field vole,Microtus montebelli" Advances in contraceptive delivery systems. (in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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