Changes in protein synthesis and phosphorylation patterns during bovine oocyte maturation in vitro.
Changes in protein synthesis and phosphorylation patterns during bovine oocyte maturation in vitro.
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DOI：
10.1530/jrf.0.0900305
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发表时间：
1990-09
期刊：
影响因子：
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通讯作者：
P. Kastrop;M. Bevers;O. Destrée;T. Kruip
中科院分区：
文献类型：
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作者：
P. Kastrop;M. Bevers;O. Destrée;T. Kruip
用[35S]蛋氨酸和[32P]正磷酸盐分别对不同时期的牛卵丘细胞复合物进行放射性标记，产生了顺序的蛋白质合成和蛋白质磷酸化模式。放射性标记的卵母细胞被评估其核状态，并单独用于凝胶电泳。蛋白质合成模式的显著变化只发生在生发囊泡破裂（GVBD）后，而保持在生发囊泡阶段的卵母细胞则表现出一致的蛋白质合成模式。在培养后8和16小时，GVBD后不久和第一次极体挤压前观察到这些变化。培养3 h后，观察到显性磷酸化蛋白条带，表观分子量为24000，两条在50000 ~ 60000之间。后一条带的分子量变化不大，与时间关系不密切。GVBD后，Mr为19000的磷酸化蛋白条带不复存在。本研究表明，在牛卵母细胞成熟过程中，蛋白质合成和磷酸化的特异性变化是程序化的。
Sequential protein synthesis and protein phosphorylation patterns were generated by radiolabelling bovine cumulus-oocyte complexes after various periods of culture with [35S]methionine and [32P]orthophosphate respectively. The radiolabelled oocytes were assessed for their nuclear status and used individually for gel electrophoresis. Marked changes in the protein synthesis patterns were observed exclusively after germinal vesicle breakdown (GVBD), whereas oocytes which remained in the germinal vesicle stage showed a consistent protein synthesis pattern. The changes were observed after 8 and 16 h or culture, shortly after GVBD and before first polar body extrusion. From 3 h of culture, dominant phosphoprotein bands with apparent molecular weights of 24,000 and two between 50,000 and 60,000 were observed. The latter bands displayed slight molecular weight changes, which were not closely time related. After GVBD, the phosphoprotein band with Mr 19,000 was no longer observed. This study demonstrates that specific changes in protein synthesis and protein phosphorylation are programmed during bovine oocyte maturation.