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首页> 外文期刊>Molecular reproduction and development >Protein kinase C and meiotic regulation in isolated mouse oocytes.
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Protein kinase C and meiotic regulation in isolated mouse oocytes.

机译：分离的小鼠卵母细胞中的蛋白激酶C和减数分裂调控。

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In this study, the possible role of protein kinase C (PKC) in mediating both positive and negative actions on meiotic maturation in isolated mouse oocytes has been examined. When cumulus cell-enclosed oocytes (CEO) were cultured for 17-18 hr in a medium containing 4 mM hypoxanthine (HX) to maintain meiotic arrest, each of the five different activators and five different antagonists of PKC stimulated germinal vesicle breakdown (GVB) in a dose-dependent fashion. One of the activators, phorbol-12-myristate 13-acetate (PMA), also triggered GVB in CEO arrested with isobutylmethylxanthine or guanosine, but not in those arrested with dibutyryl cyclic AMP. When denuded oocytes (DO) were cultured for 3hr in inhibitor-free medium, all PKC activators suppressed maturation (<10% GVB compared to 94% in controls), while the effect of PKC antagonists was negligible. Four of the five antagonists reversed the meiosis-arresting action of HX in DO. PMA transiently arrested the spontaneous maturation of both CEO and DO, with greater potency in DO. The stimulatory action of PMA in HX-arrested oocytes was dependent on cumulus cells, because meiotic induction occurred in CEO but not DO. PKC activators also preferentially stimulated cumulus expansion when compared to antagonists. A cell-cell coupling assay determined that the action of PMA on oocyte maturation was not due to a loss of metabolic coupling between the oocyte and cumulus oophorus. Finally, Western analysis demonstrated the presence of PKCs alpha, beta1, delta, and eta in both cumulus cells and oocytes, but only PKC epsilon was detected in the cumulus cells. It is concluded that direct activation of PKC in the oocyte suppresses maturation, while stimulation within cumulus cells generates a positive trigger that leads to meiotic resumption.
机译：在这项研究中，已经研究了蛋白激酶C（PKC）在介导分离的小鼠卵母细胞成熟中减数分裂成熟的正面和负面作用中的可能作用。当将卵丘细胞封闭卵母细胞（CEO）在含有4 mM次黄嘌呤（HX）的培养基中培养17-18小时以维持减数分裂阻滞时，PKC的五种不同激活剂和五种不同拮抗剂各自刺激生发囊泡分解（GVB）以剂量依赖的方式。其中一种激活剂佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）也会在被异丁基甲基黄嘌呤或鸟苷逮捕的CEO中触发GVB，但在被二丁酰基环AMP逮捕的CEO中则触发GVB。当裸露的卵母细胞（DO）在无抑制剂的培养基中培养3小时时，所有PKC激活剂均抑制成熟（GVB <10％，而对照中为94％），而PKC拮抗剂的作用可忽略不计。五个拮抗剂中的四个逆转了DO中HX的减数分裂阻滞作用。 PMA暂时阻止了CEO和DO的自发成熟，在DO中具有更大的潜力。 PMA在HX阻滞卵母细胞中的刺激作用取决于卵丘细胞，因为减数分裂诱导发生在CEO而非DO中。与拮抗剂相比，PKC激活剂还优先刺激卵丘扩张。细胞-细胞偶联测定法确定PMA对卵母细胞成熟的作用不是由于卵母细胞和卵丘之间的代谢偶联丧失。最后，Western分析表明，在卵丘细胞和卵母细胞中都存在PKCsα，beta1，δ和eta，但在卵丘细胞中仅检测到PKCε。结论是卵母细胞中PKC的直接激活抑制了成熟，而卵丘细胞内的刺激产生了导致减数分裂恢复的正触发。

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