Molecular Characterization and Polyadenylation-Regulated Expression of Cyclin B1 and Cdc2 in Porcine Oocytes and Early Parthenotes
Molecular Characterization and Polyadenylation-Regulated Expression of Cyclin B1 and Cdc2 in Porcine Oocytes and Early Parthenotes
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DOI：
10.1002/mrd.21095
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发表时间：
2010-01-01
影响因子：
2.5
通讯作者：
Kim, Nam-Hyung
中科院分区：
文献类型：
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作者：
Zhang, Ding-Xiao;Cui, Xiang-Shun;Kim, Nam-Hyung
哺乳动物卵母细胞减数分裂成熟受成熟/M期促进因子（MPF）控制，MPF是Cdc 2激酶和细胞周期蛋白B蛋白的复合物。为了更好地了解卵母细胞成熟的分子机制，我们对猪细胞周期蛋白B1和Cdc 2基因进行了表征，这两个基因在猪的组织中广泛表达。我们进一步从mRNA和蛋白水平分析了它们在猪卵母细胞体外成熟和早期胚胎发育过程中的表达谱。细胞周期蛋白B1的两种亚型，包括相同的开放阅读框架，但不同的3 '-UTR长度，被确定。细胞周期蛋白B1的转录上调后30小时的成熟，而Cdc 2的mRNA水平在成熟过程中没有变化，但在44小时急剧下降。细胞周期蛋白B1的蛋白合成增加与卵母细胞成熟。Cdc 2蛋白表达在0-18小时期间相对较低，随后在成熟的44小时内表达水平较高。Poly（A）-test PCR清楚地显示，两种细胞周期蛋白B1亚型在成熟过程中约18-24小时开始发生胞质聚腺苷酸化，而在中期II卵母细胞和孤雌激活后的胚胎发育过程中观察到Cdc 2亚型的大量去腺苷酸化和降解。来自小卵泡的猪MII卵母细胞（
Meiotic maturation of mammalian oocytes is controlled by the maturation/M-phase promotion factor (MPF), a complex of Cdc2 kinase and cyclin B protein. To better understand the molecular mechanism of oocyte maturation, we characterized porcine cyclin B1 and Cdc2 genes, both of which are widely expressed in pig tissues. We further analyzed their expression profiles during in vitro maturation of pig oocyte and early embryonic development at both the mRNA and protein level. Two isoforms of cyclin B1, comprising the same open reading frame but differing in 3'-UTR length, were identified. Cyclin B1 transcripts was up-regulated after 30 hr of maturation, while Cdc2 mRNA levels were unchanged during maturation except for a sharp decline at 44 hr. Cyclin B1 protein synthesis increased with oocyte maturation. Cdc2 protein expression was relatively low during 0-18 hr, followed by a higher level of expression up to 44 hr of maturation. Poly(A)-test PCR clearly revealed that both cyclin B1 isoforms underwent cytoplasmic polyadenylation starting around 18-24 hr during maturation, while a substantial de-adenylation and degradation of Cdc2 isoforms were observed in metaphase II oocytes and during embryo development after parthenogenetic activation. Porcine MII oocytes derived from small follicles (