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蛋白质合成抑制后牛卵母细胞中的组蛋白H1和丝裂原活化蛋白激酶活性

Histone H1 and MAP kinase activities in bovine oocytes following protein synthesis inhibition.

作者信息

Meinecke B, Janas U, Podhajsky E, Meinecke-Tillmann S

机构信息

Institute of Reproductive Medicine, School of Veterinary Medicine Hannover, Hannover, Germany.

出版信息

Reprod Domest Anim. 2001 Aug;36(3-4):183-8.

Abstract

In vitro nuclear maturation is associated with known activity profiles of the M-phase promoting factor (MPF) and the mitogen-activated protein (MAP) kinases, which are two key regulators of mitotic and meiotic cell cycles. Initiation of meiotic resumption in vitro can be prevented by cycloheximide treatment and after removal of the inhibitor germinal vesicle breakdown takes place nearly twice as fast as in untreated controls. In this study experiments were conducted in order to examine the chromosome condensation status and the dynamics of MPF and MAP kinase activities after cycloheximide treatment (10 microg/ml) of cumulus-enclosed oocytes for 17 and 24 h, respectively, and subsequent culture in inhibitor-free medium for various times. Bovine oocytes displayed variations in the degree of chromosome condensation at the end of the inhibitor treatment phase. Following removal of the inhibitor germinal vesicle breakdown occurred after 4-5 h of subsequent culture in inhibitor-free medium. MPF and MAP kinase exhibited low activities during the first 1-3 h following cycloheximide treatment. Increasing levels of enzyme activities were detected 4-7 h following cycloheximide treatment for 17 and 24 h, respectively, and subsequent culture in inhibitor-free medium. The patterns of enzyme activities corresponded with the accelerated nuclear maturation process. It can be concluded that cycloheximide treatment does not lead to a more synchronous course of nuclear maturation and that the activities of both, MPF and MAP kinase are initiated at least 2-5 h earlier in comparison with untreated oocytes.

摘要

体外核成熟与促有丝分裂因子(MPF)和丝裂原活化蛋白(MAP)激酶的已知活性谱相关,这两种蛋白是有丝分裂和减数分裂细胞周期的两个关键调节因子。体外减数分裂恢复的启动可通过环己酰亚胺处理来阻止,去除抑制剂后,生发泡破裂的发生速度几乎是未处理对照组的两倍。在本研究中,进行了实验,以检查分别用环己酰亚胺(10微克/毫升)处理卵丘包裹的卵母细胞17小时和24小时后,以及随后在无抑制剂培养基中培养不同时间后,染色体凝聚状态以及MPF和MAP激酶活性的动态变化。在抑制剂处理阶段结束时,牛卵母细胞的染色体凝聚程度存在差异。去除抑制剂后,在无抑制剂培养基中后续培养4 - 5小时后生发泡破裂发生。在环己酰亚胺处理后的最初1 - 3小时内,MPF和MAP激酶活性较低。分别在环己酰亚胺处理17小时和24小时后,以及随后在无抑制剂培养基中培养,在4 - 7小时检测到酶活性水平升高。酶活性模式与加速的核成熟过程相对应。可以得出结论,环己酰亚胺处理不会导致核成熟过程更加同步,并且与未处理的卵母细胞相比,MPF和MAP激酶的活性至少提前2 - 5小时开始。

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