Improvement of meiotic competence of follicular oocytes by artificial techniques

通过人工技术提高卵泡卵母细胞减数分裂能力

• 批准号: 61560305
• 负责人: SATO Eimei
• 金额: $ 1.34万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61560305/
• 关键词: 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

1. Improvement of meiotic competence of follicular oocytes by chemicalsOocytes 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 glycosaminoglycansMouse 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

1.化学物质提高卵泡卵母细胞减数分裂能力的研究用针和刀片机械分离法从卵巢中分离出不同大小的卵母细胞。这种方法产生了26.2 - 7.4生长和完全皱眉卵母细胞从卵巢。从卵巢中分离出的卵丘包裹的卵母细胞在含有10%胎牛血清的TCM 199中进行体外培养，直径90 m的卵母细胞均未恢复减数分裂。但在直径大于91 μ m的卵母细胞中观察到萌发囊泡破裂（GVDB）。在直径超过101 μ m的卵母细胞中观察到极体形成。约80%的121 m直径的卵母细胞能够挤出极体。在直径为101 ~ 120 m的卵母细胞中，用含有3 '，5'-二丁酰环腺苷酸的培养液孵育时，第一极体的比例增加。这些结果表明，二丁酰环腺苷3 '，5'-单磷酸能提高牛的减数分裂能力 ...更多信息 在培养中培养卵母细胞。卵巢糖胺聚糖对小鼠卵母细胞自发性变性的预防作用小鼠卵泡卵母细胞在改良Krebs-Ringer液中体外培养72 h以上，发生自发性分裂，形成带有或不带有不清晰碎片或染色质的“卵裂球”。从牛卵泡液（bFF）中分离的糖胺聚糖（GAGs）以0.25mg/ml或更高的浓度加入到悬浮培养液中，以剂量依赖的方式阻止小鼠卵母细胞自发分裂的发生。通过在dowex 1-x2上的色谱法从bFF中分离活性物质。用0.5M NaCl洗脱的组分具有最大的生物活性。从商业来源制备纯化的bFF-GAGs和透明质酸和硫酸软骨素防止了分割的发生。透明质酸比硫酸软骨素更有效。结果表明，糖胺聚糖在体外可防止卵母细胞的变性。少

项目成果

Sato,E.: Journal of Animal Science.

佐藤，E.：动物科学杂志。

佐藤英明: 実験動物. 37. 231-238 (1988)

佐藤英明：实验动物。37。231-238（1988）

佐藤英明: 遺伝. 42. 72-75 (1988)

佐藤秀明：遗传学 42. 72-75 (1988)

Sato, E.: Poultry Science. 66. 1053-1058 (1987)

佐藤，E.：家禽科学。

Sato, E.: "Biochemical transmitters regulating the arrest and resumption of meiosis in oocytes" International Review of Cytology. 106. 1-33 (1987)

Sato，E.：“调节卵母细胞减数分裂停滞和恢复的生化递质”国际细胞学评论。