蛋白激酶C在小鼠卵母细胞体外成熟和受精中的作用  

ROLES OF PROTEIN KINASE C IN MOUSE OOCYTE MATURATION AND FERTILIZATION IN VITRO

作　　者：范衡宇[1] 佟超[1] 李世文[1] 陈大元[1] 孙青原[1] 

机构地区：[1]中国科学院动物研究所计划生育生殖生物学国家重点实验室,北京100080

出　　处：《实验生物学报》2003年第1期37-42,共6页Acta Biologiae Experimentalis Sinica

基　　金：973项目(G1999055902);中国科学院创新工程项目(KSCX2-SW-303);国家杰出青年基金(No.30225010)

摘　　要：蛋白激酶是一类重要的丝/苏氨酸蛋白激酶。本实验以小鼠为实验动物,研究了PKC在卵母细胞体外成熟、活化和受精中的可能作用,及两种PKC亚型在卵母细胞中的定位。PKC激活剂PMA可以阻止CV期卵母细胞在体外恢复减数分裂,该作用可被PKC抑制剂calphostin C抵消,但不能被PLCγ抑制剂U73122或PKCδ专一性抑制剂Rottlerin所克服。Western印迹显示PKCα和βⅠ在卵母细胞发育过程中恒量表达。激光共聚焦显微术研究发现,受精或受到活化刺激后PKCα转位到卵母细胞膜上,同时皮质颗粒排放,说明PKCα可能参与调节卵皮质反应。本实验首次在小鼠中研究了PLCγ与受精的关系,发现不存在PKC对PLCγ的正反馈调节。此外,本研究还对小鼠卵巢中对PKCα和βⅠ进行了蛋白定位研究。Protein kinase C is an important family of serine/lhreorine protein kinase. In this experiment the possible roles of PKC in meiotic maturation, fertilization and egg activation were studied in mouse oocytes. Two isoforms of PKC, PKC α and βI, were localized in mouse oocytes. PKC activator PMA inhibited the resumption of meiosis in oocytes at GV stage, whose effect could be overcomed by PKC inhibitor calphostin C, neither the phospholipase C inhibitor U73122 nor the PKCδ-specific inhibitor rottlerin. The result of Western bolt indicates that both PKC α and βI express stably during oocyte development. The translocation of PKCs and cortical granule exocytosis were observed by confocal microscopy. The results show that PKC α translocate from the cytoplasm to the membrane after fertilization or drug-stimulation, accompanied by CG exocytosis. This fact suggested that PKC α might be involved in the regulation of cortical reaction in mouse oocytes. Roles of PLC γ in mouse oocyte during fertilization and localization of PKCs in mouse ovary were also investigated in this experiment. The result suggested that there did not exist a feedback regulation of PLC γ on PKC during fertilization.

关 键 词：蛋白激酶C 小鼠 卵母细胞 体外成熟 受精 作用 

分 类 号：Q55[生物学—生物化学]