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Study on Activation of Oocytes Which Are Injected With a Round Spermatid

Research Project

Project/Area Number 10671549
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Obstetrics and gynecology
Research Institutionthe University of Tokushima

Principal Investigator

YAMANO Shuji  The University of Tokushima, University Hospital, Associate Professor, 医学部・附属病院, 助教授 (30166772)

Co-Investigator(Kenkyū-buntansha) KOMATSU Junko  The University of Tokushima, University Hospital, medical staff, 医学部・附属病院, 医員(臨床)
YAMASITA Mizuho  The University of Tokushima, University Hospital, medical staff, 医学部・附属病院, 医員(臨床)
NAKAGAWA Koji  The University of Tokushima, University Hospital, Research Associate, 医学部・附属病院, 助手 (40314869)
檜尾 健二  徳島大学, 医学部付属病院, 医員(臨床)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordsfertilization / oocyte activation / parthenogenone / MPF / MAPK / 卵活性化 / 顕微授精 / 卵細胞質内精子注入法 / 染色体
Research Abstract

Recent reports suggest that when unfertilized oocytes with a spermatozoon after intracytoplasmic sperm injection (ICSI) are properly activated, the activated oocytes develop normally like fertilized oocytes. The aim of the study is to develop the activation method which is able to produce embryos with normal karyotype from unfertilized oocytes with a spermatozoon, the activation method which produces one pronucleus with extrusion of the second polar body in oocytes without a spermatozoon is necessary.
1) When human unfertilized oocytes 22 h after insemination were treated with 5 M Calcium ionophore for 5 min and incubated in HTF medium with 10 g/ml puromycin for 5 h, the activation rate was about 90% and the proportion of parthenogenones displaying one pronucleus (PN) with extrusion of the second polar body (PB) was about 80%.
2) When mouse ovulated oocytes were treated with 5 M Calcium ionophore for 5 min and incubated in HTF medium with 10 g/ml puromycin for 4 h, the activation rate and the proportion of parthenogenones displaying one PN with extrusion of the second PB were similar to those of human aged oocytes treated with the same method.
3) When human unfertilized oocytes following ICSI were activated by the method, two PN were formed with extrusion of the second PB in only 30% of the parthenogenones and 64% of the parthenogenones with 2PN2PB cleaved. Four clcaved parthenogenones showed normal karyotype.
4) When maturation promoting facor (MPF) and mitogen activated protein kinase (MAPK) in mouse oocytes treated with the method were measured. MPF significantly decreased first in a time-dependent manner, and sequentially MAPK decreased.
These findings suggest that the activation method decreased MPF and MAPK like fertilized oocytes, and extrudes the second PB and forms a female PN. However, puromycin may suppress the formation of a male PN.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Tokumura A. et al.: "Production of lysophosphatidic acid by lysophospholipase D in follicular fluids of patients for in vitro fertilization"Biol Reprod. 61. 195-199 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kunikata K. et al.: "Effect of lysophosphatidic acid on the ovum transport in mouse oviducts"Life Science. 69. 833-840 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakagawa K. et al.: "Quality of embryo does not affect the implantation rate of in vitro fertilization - embryo transfer in infertile women with antisperm antibody"Fertil Steril. 72. 1055-1060 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yamano S. et al.: "Fertilization failure and oocyte activation"Journal of Medical Investigation. 47. 1-8 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakasaka H, et al.: "Effective activation method with A23187 and puromycin to produce haploid parthenogenones from freshly ovulated mouse oocytes"Zygote. 8. 203-208 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Quan S. et al.: "Effects of preservation of mouse spermatozoa in electrolite-free solution at 4℃ on the outcome of mouse in vitro fertilization"J Assist Reprod Genet. 17. 384-388 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakagawa K. et al.: "A combination of Ca ionophore and puromycin effectively produces human parthenogenones with one haploid pronucleus"Zygote. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Okitu O. et al.: "Activation of bovine oocytes matured in vitro by injection of bovine, human spermatozoa or their cytosolic fractions"Zygote. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takikawa M. et al.: "Evaluation of two sperm antigens, rSMP-Band YWK-II as targets for immunocontraception"Zygote. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tokumura, A., Yamano, S., Aono, T.: "Production of lysophosphatidic acid by lysophospholipase D in follicular fluids of patients for in vitro fertilization"Biol Reprod. 61. 195-199 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kunikata, K., Yamano, S., Tokumura, A., Aono, T.: "Effect of lysophosphatidic acid on the ovum transport in mouse oviducts"Life Science. 69. 833-840 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakagawa, K., Yamano, S., Kamada, M., Hinokio, K., Maegawa, M., Aono, T.: "Quality of embryo does not affect the implantation rate of in vitro fertilization embryo transfer in infertile women with antisperm antibody"Fertil Steril. 72. 1055-1060 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yamano, S., Nakagawa, K., Nakasaka, H., Aono, T.: "Fertilization failure and oocyte activation"Journal of Medical Investigation. 47. 1-8 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakasaka, H., Yamano, S., Hinokio, K., Nakagawa, K., Yoshizawa, M., Aono, T.: "Effective activation method with A23187 and puromycin to produce haploid parthenogenones from freshly ovulated mouse oocytes"Zygote. 8. 203-208 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Quan, S., Yamano, S., Nakasaka, H., Hinokio, K., Nakagawa, K., Aono, T.: "Effects of preservation of mouse spermatozoa in electrolite-free solution at 4℃ on the outcome of mouse, in vitro fertilization"J Assist Reprod Genet. 17. 384-388 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takagawa, K., Yamano, S., Nakasaka, H., Hinokio, K., Yoshizawa, M., Aono, T.: "A combination of Ca ionophore and puromycin effectively produces human parthenogenones with noe haploid pronucleus"Zygote. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Okitu, O., Yamano, S., Aono, T.: "Activation of bovine oocytes matured in vitro by injection of bovine, human spermatozoa or their cytosolic fractions"Zygote. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takikawa M., Kamada M., Maegawa M., Yamano S., Irahara M., Aono T., Futaki S., Ohmoto Y., Koide S.S.: "Evaluation of two sperm antigens, rSMP-B and YWK-II, as targets for immuno-contraception"Zygote. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yamano S: "Fertilization failure and Oocyte activation"J Med.invest.. 47. 1-8 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nakasaka H: "Effective activation method with A23187 and puromycin to produce haploid parthenogenones from freshly ovulated mouse oocytes"Zygote. (in press).

    • Related Report
      1999 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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