REGULATION OF HSP70 MESSENGER-RNA LEVELS DURING OOCYTE MATURATION AND ZYGOTIC GENE ACTIVATION IN THE MOUSE
REGULATION OF HSP70 MESSENGER-RNA LEVELS DURING OOCYTE MATURATION AND ZYGOTIC GENE ACTIVATION IN THE MOUSE
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DOI：
10.1016/0012-1606(91)90423-z
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发表时间：
1991-04-01
影响因子：
2.7
通讯作者：
SCHULTZ, RM
中科院分区：
文献类型：
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作者：
MANEJWALA, FM;LOGAN, CY;SCHULTZ, RM
camp依赖性蛋白激酶催化的蛋白磷酸化与双细胞小鼠胚胎基因组的转录激活有关，而热休克蛋白（hsp70）已被确定为合子基因激活的首批产物之一。利用逆转录聚合酶链反应，我们分析了hsp70 mRNA在卵母细胞成熟和早期胚胎发生过程中的相对变化。我们报道，hsp70 mRNA的数量在生发囊泡破裂后减少，而抑制生发囊泡破裂抑制这种成熟相关的减少。hsp70 mRNA的表达量在单细胞期和双细胞期增加。α-amanitin或camp依赖性蛋白激酶抑制剂H-8均可抑制这种增加；同样浓度的H-7对hsp70 mRNA相对量的增加几乎没有抑制作用，而H-7是一种更有效的蛋白激酶C抑制剂。最后，在G2后期的单细胞胚胎中添加环己亚胺，既没有抑制双细胞阶段的分裂，也没有抑制hsp70 mRNA相对量的增加。这些结果加强了先前提出的蛋白磷酸化参与双细胞小鼠胚胎的合子基因激活。
Protein phosphorylation catalyzed by the cAMP-dependent protein kinase is implicated in transcriptional activation of the embryonic genome in the two-cell mouse embryo, while heat shock protein (hsp70) has been identified as one of the first products of zygotic gene activation. Using reverse transcription-polymerase chain reaction we have analyzed relative changes in the amount of hsp70 mRNA during oocyte maturation and early embryogenesis. We report that the amount of hsp70 mRNA decreases after germinal vesicle breakdown, while inhibiting germinal vesicle breakdown inhibits this maturation-associated decrease. The amount of hsp70 mRNA increases between the one- and two-cell stages. This increase is inhibited by either α-amanitin or the cAMP-dependent protein kinase inhibitor H-8; the same concentration of H-7, which is a more potent inhibitor of protein kinase C, has little inhibitory effect on this increase in the relative amount of hsp70 mRNA. Last, addition of cycloheximide to one-cell embryos late in G2 inhibits neither cleavage to the two-cell stage nor the increase in the relative amount of hsp70 mRNA. These results strengthen the previous proposal that protein phosphorylation is involved in zygotic gene activation in the two-cell mouse embryo.