磷酸二酯酶亚型抑制剂与细胞周期蛋白抑制剂对绵羊卵母细胞体外成熟的影响  

Effect of the inhibitor of cdk and specific phosphodiesterase isoenzyme inhibitors on ovine oocytes meiotic maturation in vitro

作　　者：周波 洪海燕[2] 旭日干[1] 夏国良[2] 

机构地区：[1]内蒙古大学哺乳动物生殖生物学与生物技术教育部重点实验室,呼和浩特010021 [2]中国农业大学生物学院,北京100094

出　　处：《中国农业大学学报》2005年第2期1-5,共5页Journal of China Agricultural University

基　　金：教育部重点实验室基金资助项目

摘　　要：选用磷酸二酯酶3(PDE3)特异性抑制剂Milrinone和PDE4特异性抑制剂Rolipram以及细胞周期蛋白依赖性激酶(cdk)抑制剂Roscovitine(ROSC)对绵羊卵丘卵母细胞复合物(COCs)的体外自发成熟进行研究,拟建立一种模拟体内卵泡环境的绵羊卵母细胞体外培养模型。以M 199为基础培养液,绵羊COCs在含有5 0 μmol/L的Milrinone、Rolipram、ROSC或不含上述任何抑制剂的培养液中分别培养2 4h。COCs在含有5 0 μmol/LROSC的培养液中培养2 4h ,转移到含有2 0IU/LFSH的培养液继续培养2 4h检查核相。结果表明:COCs在M 199基础培养液和含有Rolipram的培养液中都可以自发恢复减数分裂,但是在含有Milrinone或ROSC的培养液中减数分裂受到阻滞;并且ROSC对绵羊COCs体外自发成熟的抑制作用要远高于Milrinone(P <0. 0 5 )。当COCs从ROSC转移到含有2 0IU/LFSH的培养液继续培养2 4h后,90 . 4 %卵母细胞恢复到第2次减数分裂中期(MⅡ)。上述结果表明PDE3和cdk在绵羊减数分裂恢复中具有重要的作用。The present works studied the effect of inhibitor of cdk and specific inhibitors of phosphodiesterase on ovine oocyte maturation. Milrinone and Rolipram are specific PDE inhibitors and Roscovitine（ROSC） is a selective inhibitors of the cyclin-dependent kinase. These chemicals are selected to evaluated the effect on meiotic resumption of ovine oocytes for 24 h in vitro. Oocytes that treated with ROSC for 24 h and subsequently allowed to undergo maturation for 24 h in the absence of the inhibitor media but have FSH（20 IU/L）. Immature ovine cumulus oocyte complexes were recovered from ovaries collected at the abattoir. The doses of inhibitors was .50μmol/L in medium. At the end of the incubation, COC were denuded and oocytes werethen fixed and classified according to the status of nuclear maturation. The results revealed that both PDE.3 inhibitor Milrinone and cdk inhibitor ROSC prevented oocyte spontaneous meiotic maturation, in contrast, the PDE4 inhibitor Rolipram had no effect. And ROSC have a high effect to maintain ovine oocytes in meiotic arrest than Milrinone. These results suggests that type 3 PDE and cyclin-dependent kinase have an important role in meiotic resumption of ovine oocytes, but the mechanisms underlying these effects are different and require further characterization.

关 键 词：减数分裂 绵羊卵母细胞 磷酸二酯酶 细胞周期蛋白依赖性激酶 

分 类 号：Q492.5[生物学—生理学]