PROTEIN-SYNTHESIS AND PHOSPHORYLATION PATTERNS OF BOVINE OOCYTES MATURING INVIVO
PROTEIN-SYNTHESIS AND PHOSPHORYLATION PATTERNS OF BOVINE OOCYTES MATURING INVIVO
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DOI：
10.1002/mrd.1080290309
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发表时间：
1991-07-01
影响因子：
2.5
通讯作者：
KRUIP, TAM
中科院分区：
文献类型：
--
作者：
KASTROP, PMM;BEVERS, MM;KRUIP, TAM
为了研究体内牛卵母细胞成熟过程中的蛋白质合成和磷酸化，在排卵前黄体生成素（LH）峰值后连续收集卵母细胞。因此，在黄体生成素峰值达到最大值后的3 ~ 20小时内，进行超排卵治疗的母牛卵巢切除。随后，选择非闭锁卵泡直径为10 mm的卵母细胞，用s- 35-蛋氨酸或p -32-正磷酸盐放射标记3小时，并单独制备凝胶电泳。蛋白质合成模式的改变与生发囊泡破裂（GVBD）一致。在随后的成熟期或与第一极体挤压相吻合的时期没有发现任何变化。此外。蛋白磷酸化模式在GVBD周围表现出显著差异。特别是，一个19 kDa的磷酸化蛋白带和两个明显分子量在50和60 kDa之间的重度磷酸化蛋白存在于萌发囊泡期的卵母细胞模式中。这些结果与先前在体外成熟过程中获得的数据有关。
To investigate protein synthesis and phosphorylation during bovine oocyte maturation in vivo, oocytes were collected at consecutive times after the preovulatory luteinizing hormone (LH) peak. Therefore, heifers treated for superovulation were ovariectomized between 3 and 20 h after the maximum of the LH peak. Subsequently, cumulus-enclosed oocytes, selected from nonatretic follicles > 10 mm, were radiolabeled with S-35-Methionine or P-32-orthophosphate for 3 h and individually prepared for gel electrophoresis. Changes in the protein synthesis patterns were observed coinciding with germinal vesicle breakdown (GVBD). No changes were detected during the ensuing maturation period or coinciding with the extrusion of the first polar body. In addition. the protein phosphorylation patterns exhibited striking differences around GVBD. In particular, a phosphoprotein band of 19 kDa and the two heavily phosphorylated proteins with apparent molecular weights between 50 and 60 kDa were present in patterns of oocytes in the germinal vesicle stage. The results are discussed in relation to previous data obtained during maturation in vitro.