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Measurement of Glucose 6-Phosphate Dehydrogenase Activity in Bovine Oocytes and Embryos Using Qualitative Fluoresence Microscopy

Research Project

Project/Area Number 01560299
Research Category

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

Allocation TypeSingle-year Grants
Research Field 畜産学(含草地学)
Research InstitutionFaculty of Agriculture, Tokyo University of Agriculture

Principal Investigator

IWASAKI Setsuo  東京農業大学, 農学部, 講師 (50184867)

Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsX chromosome / Enzyme / Bovine / Embryo / In vitro fertilization
Research Abstract

The activities of glucose 6-phosphate dehydrogenase (G6PDH) of the follicular oocytes collected from bovine ovaries and the embryos fertilized in vitro were measured using fluorescence microscope with photomultiplier and photometer attachments. The response for the fluorescence of NADPH was found to be linear, with correlation coefficiency of 0.997 and the limit of detection was 0.5mM.
The G6PDH activity of intact oocytes could not be measured directly. Then the zona pellucida free follicular oocytes in extraction solution were frozen in LN_2, thawed and centrifuged and the G6PDH activity of their supernatant could be measured. Using this method, the G6PDH activities of oocytesat germinal vesicle and second metaphase stages and embryos at two-cell to morulae stages were determined. The G6PDH activities were 0.67 to 0.84 nmol NADPH generated/embryo/h at germinal vesicle stage to four-cell stage and the activity was decreased thereafter. There was no activity at morulae stage.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] 岩崎 説雄,塩谷 康生,中原 達夫: "顕微測光により牛卵子および胚のグルコ-ス6リン酸脱水素酵素活性の微量測定" 家畜繁殖学雑誌.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Setsuo Iwasaki, Yasuo Shioya and Tatsuo Nakahara: "Measurement of glucose 6-phosphate dehydrogenase activity in bovine oocytes and embryos using quantitative fluorescence microscopy." Japanese Journal of Animal Reproduction. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 岩崎 説雄・塩谷 康生・中原 達夫: "顕微測光による牛卵子および胚のグルコ-ス6リン酸脱水素酵素活性の微量測定" 家畜繁殖学雑誌.

    • Related Report
      1990 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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