本文選題：周期依賴性蛋白激酶抑制劑(roscovitine + ROS)��； 參考：《中國獸醫(yī)學(xué)報(bào)》2017年07期

【摘要】：利用周期依賴性蛋白激酶抑制劑(roscovitine,ROS)誘導(dǎo)體外培養(yǎng)山羊的卵母細(xì)胞核質(zhì)同步成熟。用40μmol/L的ROS預(yù)處理山羊卵母細(xì)胞8h后轉(zhuǎn)入常規(guī)成熟培養(yǎng)8,12,16h,記為(8+8,8+12和8+16),統(tǒng)計(jì)0,8,8+8,8+12,8+16h卵母細(xì)胞核成熟情況;用彗星試驗(yàn)檢測(cè)8+16h時(shí)卵母細(xì)胞的老化程度;在激光共聚焦顯微鏡(Olympus,Tokyo,Japan)下觀察8+16h時(shí)卵母細(xì)胞的皮質(zhì)顆粒遷移情況;對(duì)體外培養(yǎng)8+16h卵母細(xì)胞進(jìn)行孤雌激活和體外培養(yǎng),統(tǒng)計(jì)孤雌激活胚的體外發(fā)育情況;以上試驗(yàn)都以常規(guī)培養(yǎng)相同時(shí)間為對(duì)照。結(jié)果顯示:山羊卵母細(xì)胞在體外核成熟抑制培養(yǎng)8h時(shí),處于GV期的卵母細(xì)胞為59.16%,與對(duì)照組(25.75%)差異顯著(P0.05);8+8h和8+12h時(shí),到達(dá)MⅡ期的卵母細(xì)胞的比率為19.75%和28.93%,與對(duì)照組(40.78%,53.10%)差異顯著;8+16h時(shí),到達(dá)MⅡ期的卵母細(xì)胞比率(73.20%)與對(duì)照組(74.36%)無顯著性差異(P0.05);(8+16h)組有老化傾向的卵母細(xì)胞的比率(21.67%)顯著低于對(duì)照組(35.67%);(8+16h)組卵母細(xì)胞的皮質(zhì)顆粒完全遷移率(81.84%)與對(duì)照組(78.89%)差異不顯著;(8+16h)組卵母細(xì)胞孤雌胚的囊胚發(fā)育率(38.60%)高于對(duì)照組(32.80%)(P0.05)。結(jié)果表明:成熟液中添加40μmol/L ROS對(duì)山羊卵母細(xì)胞短時(shí)間(8h)核成熟抑制培養(yǎng),然后轉(zhuǎn)入常規(guī)成熟培養(yǎng)16h,即不改變山羊卵母細(xì)胞體外成熟的時(shí)間(24h),可有效延遲部分卵母細(xì)胞生發(fā)泡破裂(GVBD)的發(fā)生,不影響山羊卵母細(xì)胞核質(zhì)的成熟,不降低MⅡ期的卵母細(xì)胞比率,可以降低有老化傾向卵母細(xì)胞的比率,提高山羊卵母細(xì)胞的后續(xù)發(fā)育能力。
[Abstract]:Cytoplasmic maturation of goat oocytes in vitro was induced by cycle-dependent protein kinase inhibitor roscovitine ros.Goat oocytes were pretreated with 40 渭 mol/L ROS for 8 h and then transferred into normal maturation culture for 812 h and 8 16 h respectively. The nuclear maturation of goat oocytes was measured by comet assay at 8 16h.The migration of cortical particles of oocytes was observed under the confocal microscope (Olympus Olympus TokyoJapan) for 8 16h, the parthenogenetic activation and in vitro culture of oocytes were carried out for 8 16h, and the in vitro development of parthenogenetic activated embryos was counted.All the above experiments were controlled by conventional culture for the same time.The results showed that when goat oocytes were cultured in vitro for 8 h, the number of oocytes in GV phase was 59.16 and significantly different from that of the control group (P 0.05) at 8 h and 8 12 h.The ratio of oocytes to M 鈪，

本文編號(hào)：1741888