蛋白激酶Cα在小鼠孤雌及四倍体胚胎早期发育过程中的分布和作用  

Location and role of protein kinase Cα in parthenogenetic and tetraploid preimplantation embryonic development in mouse

作　　者：陈雅隽[1] 申景岭[1] 冯秀清[1] 单智焱[1] 阎晓飞[1] 董建将[1] 钟淑琦[1] 雷蕾[1] 

机构地区：[1]哈尔滨医科大学组织学与胚胎学教研室,哈尔滨150081

出　　处：《生理学报》2008年第1期105-112,共8页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China (No. 30671025;the Science Foundations fromEducation Department of Heilongjiang Province (No. 1151hz031);Health Department of Heilongjiang Province (No. 2006-271)

摘　　要：为了观察蛋白激酶Cα(protein kinase Cα,PKCα)在昆明白小鼠受精卵、孤雌激活和四倍体胚胎早期发育阶段的亚细胞定位和致密化进程中的表达变化,本实验利用免疫荧光化学染色与激光共聚焦显微镜观察相结合的方法,对受精卵、孤雌激活和四倍体胚胎早期发育阶段PKCα的表达进行了定位观察,并利用Westernblot对三组胚胎致密化进程中PKCα的表达进行定量分析。结果显示,PKCα在上述三组胚胎发育的2-细胞期至囊胚期均有表达,虽然不同胚胎PKCα的分布在同一发育阶段存在差异,却表现出在各胚胎期主要分布于卵裂球核染色质内,以及在胚胎致密化开始,PKCα在卵裂球连接处发生重新分布的共同特点。此外,三组胚胎PKCα在致密化进程中的表达呈升高趋势,即致密化后的表达高于致密化前。结果表明,PKCα对胚胎致密化的调节具有重要作用,其在8-细胞/4-细胞期的重新分布是胚胎进入桑椹胚期的必然事件,是胚胎致密化的前提,同时伴随蛋白表达增多。此外,PKCα在囊胚期发生了植入前的第二次重新分布。PKCα在三组胚胎各发育阶段表达情况各不相同,它对小鼠胚胎发育的影响体现在整个早期发育阶段。PKCα在小鼠受精卵早期发育阶段的两次重新分布可能与在致密化开始时启动的细胞黏附事件存在某种必然联系。Protein kinase C （PKC） is a critical molecule in cellular signal transduction in mammals. It is involved in many biological processes in embryonic development, including nuclear remodeling, cell cycle adjustment and cellular polarity regulation. The present study aimed to observe the location of PKCα, an important isozyme of PKC, in fertilized, parthenogenetic and tetraploid preimplantation embryos, and compare the expression of PKCα during embryonic compaction in Kunming mice. The location of PKCα was detected by immunochemistry and laser confocal microscopy. Western blot was performed to quantify PKCα expression during embryonic compaction in the three kinds of embryos. In the experiment, fertilized embryos were flushed from oviduct or uterus at 45, 52, 69, 76 and 93 h after injection of human chorionic gonadotrophin （hCG）; parthenogenetic embryos were collected by SrCl2 activation of oocytes for 6 h; and tetraploid embryos were produced by electrofusion of 2-cell embryos. Embryos were fixed at different developmental stages for immunofluorescent staining. 8-cell/4-cell embryos and morula were lysed for Western blot. The results showed that PKCα had similar location pattern in different embryos. It was distributed mainly in the nuclear aggregating around chromatin at different developmental stages. However, PKCα expressed strongly in the interphase than in mitotic blastomere. Before embryonic compaction, PKCα was localized at the blastomere boundary. At late blastocyst stage of fertilized embryos, PKCα was localized only in the polar trophoblast, but not in other trophoblast. At late stage of pathenogenetic blastocyst, there was no clear PKCα signal in the polar trophoblast. Tetraploid embryos had larger blastomere than other embryos and compacted after 4-cell stage, but not after 8-cell stage. Meanwhile, there was PKCα signal at the blastomere boundary at 4-cell stage. Our results showed that the expression of PKCα lasted through all the preimplantation stage. Although there were different e

关 键 词：蛋白激酶CΑ 孤雌胚胎 四倍体胚胎 致密化 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]