ROS诱导山羊卵母细胞核质同步成熟

发布时间：2018-04-12 23:19

  本文选题：周期依赖性蛋白激酶抑制剂(roscovitine + ROS)　； 参考：《中国兽医学报》2017年07期

【摘要】：利用周期依赖性蛋白激酶抑制剂(roscovitine,ROS)诱导体外培养山羊的卵母细胞核质同步成熟。用40μmol/L的ROS预处理山羊卵母细胞8h后转入常规成熟培养8,12,16h,记为(8+8,8+12和8+16),统计0,8,8+8,8+12,8+16h卵母细胞核成熟情况;用彗星试验检测8+16h时卵母细胞的老化程度;在激光共聚焦显微镜(Olympus,Tokyo,Japan)下观察8+16h时卵母细胞的皮质颗粒迁移情况;对体外培养8+16h卵母细胞进行孤雌激活和体外培养,统计孤雌激活胚的体外发育情况;以上试验都以常规培养相同时间为对照。结果显示:山羊卵母细胞在体外核成熟抑制培养8h时,处于GV期的卵母细胞为59.16%,与对照组(25.75%)差异显著(P0.05);8+8h和8+12h时,到达MⅡ期的卵母细胞的比率为19.75%和28.93%,与对照组(40.78%,53.10%)差异显著;8+16h时,到达MⅡ期的卵母细胞比率(73.20%)与对照组(74.36%)无显著性差异(P0.05);(8+16h)组有老化倾向的卵母细胞的比率(21.67%)显著低于对照组(35.67%);(8+16h)组卵母细胞的皮质颗粒完全迁移率(81.84%)与对照组(78.89%)差异不显著;(8+16h)组卵母细胞孤雌胚的囊胚发育率(38.60%)高于对照组(32.80%)(P0.05)。结果表明:成熟液中添加40μmol/L ROS对山羊卵母细胞短时间(8h)核成熟抑制培养,然后转入常规成熟培养16h,即不改变山羊卵母细胞体外成熟的时间(24h),可有效延迟部分卵母细胞生发泡破裂(GVBD)的发生,不影响山羊卵母细胞核质的成熟,不降低MⅡ期的卵母细胞比率,可以降低有老化倾向卵母细胞的比率,提高山羊卵母细胞的后续发育能力。
[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，

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