| Project/Area Number |
61560305
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
畜産学(含草地学)
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| Research Institution | Kyoto University |
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Principal Investigator |
SATO Eimei Faculty of Agriculture, Kyoto University, 農学部, 助教授 (80093243)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥100,000 (Direct Cost: ¥100,000)
Fiscal Year 1987: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1986: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Mouse / Cattle / Oocyte / cAMP / サイクリックAMP / ヒアルロン酸 / 卵胞卵 / 卵の成熟 / 卵の分離 / 卵の退行 / 卵子成熟 / 卵子成熟調節因子 / 卵子退行 / 卵子退行遅延因子 |
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| Research Abstract |
1. Improvement of meiotic competence of follicular oocytes by chemicals
Oocytes of various sizes were isolated from ovaries by mechanical dissection using the gauge-needle and razor blade. This method yielded 26.2 7.4 growing and fully frown oocytes from an ovary. Cumulus-enclosed oocytes dissociated from the ovaries were cultured in vitro in a TCM 199 containing 10% fetal calf serum, none of the 90 m-diameter oocytes resumed meiosis. However germinal vesicle breakdown (GVDB) was observed in oocytes of which diameter exceeded 91 m. Polar body formation was observed in oocytes which have the diameter exceeding 101 m. About 80% of the 121 m-diameter oocytes were able to extrude the polar body. The percentage of oocytes with first polar body increased when incubated in the medium containing dibutyryl cyclic adenosine 3',5'-monophosphate in 101-to 120 m-diameter oocytes. These observations indicate that dibutyryl cyclic adenosine 3',5'-monophosphate can improve the meiotic competence of bov
… More
ine oocytes in culture.
2. Prevention of soontaneous degeneration of mouse oocytes in culture by ovarian glycosaminoglycans
Mouse follicular oocytes cultured for more than 72 h in modified Krebs-Ringer solution in vitro undergo spontaneous segmentation to form "blastomeres" with or without unclear fragments or chromatin. Glycosaminoglycans(GAGs) isolated from bovine follicular fluid(bFF) and added to the suspending medium at concentrations of 0.25mg/ml or greater prevented the occurence of spontaneous segmentation in isolated cumulus-free mouse oocytes in a dose-dependent manner. The active material was isolated from bFF by chromatography on dowex 1-x2. The fraction eluted with 0.5 M NaCl plssessed the greatest biological activity. Preparations of purified bFF-GAGs and hyaluronic acid and chondroitin sulfate from a commercial soutce prevented the occurence of segmentation. Hyaluronic acid was more potent than chondroitin sulfate. The results indicate that GAGs prevent degeneration of oocyte in vitro. Less
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