PDE3与亮甲酚蓝在绵羊卵母细胞体外培养中的应用研究  

Application of PDE3 and Brilliant Cresyl Blue in in-vitro Culture of Sheep Oocyte

作　　者：汪立芹[1,2] 王静 金贤华[1,2] 陈博[1,2] 黄俊成[1,2] 

机构地区：[1]新疆动物生物技术重点开放实验室,新疆乌鲁木齐830000 [2]农业部草食家畜繁育生物技术重点开放实验室,新疆乌鲁木齐830000

出　　处：《安徽农业科学》2011年第25期15408-15409,15553,共3页Journal of Anhui Agricultural Sciences

基　　金：新疆自治区自然科学基金项目(200821182);新疆自治区科技攻关项目(200841122);新疆自治区科技计划项目(200711104);国家转基因重大专项

摘　　要：[目的]探讨磷酸二酯酶3(PDE3)特异性抑制剂Milrinone与亮甲酚蓝染色液(BCB)相结合在绵羊卵母细胞体外培养中的应用。[方法]比较了BCB对绵羊卵母细胞的筛选与传统的形态分级之间的差异,以及对胚胎后期发育的影响;研究了Milrinone对绵羊卵母细胞的最佳抑制时间,并用于BCB-卵母细胞体外培养。[结果]A、B级卵母细胞中BCB+卵母细胞的比例为64.42%,极显著高于C级卵母细胞的17.00%;BCB+卵母细胞的成熟率(86.16%)、卵裂率(85.29%)、囊胚率(34.40%)分别极显著高于BCB-卵母细胞(50.94%、36.19%和6.73%);6 h是Milrinone对绵羊卵母细胞的最佳抑制时间,该试验组体外培养效率显著高于其他组;Milrinone对BCB-卵母细胞抑制培养6 h,极显著提高了体外胚胎发育率。[结论]为提高绵羊卵母细胞体外培养效率提供了依据。[Objective] The aim of this study was to increase the viability of sheep oocytes in vitro by using phosphodiesterase type 3（PDE 3） inhibitors milrinone and brilliant cresyl blue（BCB） staining.[Method] The quality between BCB tested and morphologically selected oocytes,as well as the embryo development was compared;and then suitable inhibitive time of milrinone to sheep oocytes in vitro was studied and used in BCB-oocytes for in vitro embryo production（IVEP）.[Result] The BCB＋ oocytes percentage in A-and B-level sheep oocytes was 64.42%,which was extremely significantly higher than that in C-level（17.0%）.The maturing rate,cleavage rate and blastocyst rate of BCB＋ oocytes（86.16%,85.29% and 34.40%） of was significantly higher than those of BCB-oocytes（50.94%,36.19% and 6.73%）.The best time for PDE 3 inhibitor delaying the sheep oocyte mature in vitro was 6 h.In addition,the rate of embryo development in vitro could be significantly increased by inhibiting the BCB-oocytes for 6 h with Milrinone.[Conclusion] The study will provide reference for improving the efficiency of sheep oocytes culture in vitro.

关 键 词：磷酸二酯酶3 亮甲酚蓝 MILRINONE 绵羊 卵母细胞 

分 类 号：Q813.7[生物学—生物工程]