Marangos Petros, Carroll John
Department of Physiology, University College London, Gower Street, London WC1E 6BT, UK.
Reproduction. 2004 Aug;128(2):153-62. doi: 10.1530/rep.1.00192.
Cdk1-cyclin B1 kinase activity drives oocytes through meiotic maturation. It is regulated by the phosphorylation status of cdk1 and by its spatial organisation. Here we used a cyclin B1-green fluorescent protein (GFP) fusion protein to examine the dynamics of cdk1-cyclin B1 distribution during meiosis I (MI) in living mouse oocytes. Microinjection of cyclin B1-GFP accelerated germinal vesicle breakdown (GVBD) and, as previously described, overrides cAMP-mediated meiotic arrest. GVBD was pre-empted by a translocation of cyclin B1-GFP from the cytoplasm to the germinal vesicle (GV). After nuclear accumulation, cyclin B1-GFP localised to the chromatin. The localisation of cyclin B1-GFP is governed by nuclear import and export. In GV intact oocytes, cyclin export was demonstrated by showing that cyclin B1-GFP injected into the GV is exported to the cytoplasm while a similar size dextran is retained. Import was revealed by the finding that cyclin B1-GFP accumulated in the GV when export was inhibited using leptomycin B. These studies show that GVBD in mouse oocytes is sensitive to cyclin B1 abundance and that the changes in distribution of cyclin B1 contribute to progression through MI.
细胞周期蛋白依赖性激酶1(Cdk1)-细胞周期蛋白B1激酶活性驱动卵母细胞进行减数分裂成熟。它受Cdk1的磷酸化状态及其空间组织的调节。在此,我们使用细胞周期蛋白B1-绿色荧光蛋白(GFP)融合蛋白来研究活的小鼠卵母细胞减数分裂I(MI)期间Cdk1-细胞周期蛋白B1的分布动态。显微注射细胞周期蛋白B1-GFP加速了生发泡破裂(GVBD),并且如先前所述,克服了cAMP介导的减数分裂阻滞。细胞周期蛋白B1-GFP从细胞质易位到生发泡(GV)可抢先引发GVBD。核积累后,细胞周期蛋白B1-GFP定位于染色质。细胞周期蛋白B1-GFP的定位受核输入和输出的控制。在完整的GV卵母细胞中,通过显示注射到GV中的细胞周期蛋白B1-GFP被输出到细胞质而类似大小的葡聚糖被保留,证明了细胞周期蛋白的输出。当使用雷帕霉素B抑制输出时,细胞周期蛋白B1-GFP在GV中积累,这揭示了输入。这些研究表明,小鼠卵母细胞中的GVBD对细胞周期蛋白B1的丰度敏感,并且细胞周期蛋白B1分布的变化有助于通过MI的进程。