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热带爪蟾卵母细胞减数分裂成熟过程中MPF和MAPK活性的表征

Characterization of MPF and MAPK activities during meiotic maturation of Xenopus tropicalis oocytes.

作者信息

Bodart Jean-Francois L, Gutierrez Davina V, Nebreda Angel R, Buckner Bree D, Resau James R, Duesbery Nicholas S

机构信息

Van Andel Research Institute, Laboratory of Developmental Cell Biology, Special Program in Analytical, Cellular, and Molecular Microscopy, 333 Bostwick NE, Grand Rapids, Michigan 49503, USA.

出版信息

Dev Biol. 2002 May 15;245(2):348-61. doi: 10.1006/dbio.2002.0647.

Abstract

Resumption of meiosis in oocytes of Xenopus tropicalis required translation but not transcription, and was marked by the appearance of a white spot and a dark ring, coincident with entry into metaphase I and the onset of anaphase I, respectively. Cyclin B(2)/p34(cdc2) activity increased prior to the first meiotic division, declined at the onset of anaphase I, and subsequently increased again. The capacity of egg cytoplasm to induce germinal vesicle breakdown (GVBD) was inhibited by cycloheximide, despite the fact that these oocytes contained cyclin B(2)/p34(cdc2) complexes. However, cycloheximide-treated oocytes underwent GVBD following injection of constitutively active mitogen-activated protein kinase (MAPK) kinase 2 (MEK2), p33(Ringo), or Delta 90 cyclin B. MAPK activity increased just prior to the first meiotic division and remained stable thereafter. Although injection of constitutively active MEK2 induced GVBD, treatment with the MEK inhibitors U0126 or anthrax lethal factor delayed GVBD and prevented spindle formation. Interestingly, the ability of egg cytoplasm to induce GVBD was unaffected by the inhibition of MEK activity. Our results indicate that the synthesis of a novel or short-lived protein(s) which acts in a MEK-independent fashion is required in order for egg cytoplasm to induce GVBD in X. tropicalis oocytes.

摘要

热带爪蟾卵母细胞减数分裂的恢复需要翻译但不需要转录,其标志是出现一个白色斑点和一个深色环,分别与进入减数第一次分裂中期和后期开始同时发生。细胞周期蛋白B(2)/p34(cdc2)活性在第一次减数分裂前增加,在后期开始时下降,随后再次增加。尽管这些卵母细胞含有细胞周期蛋白B(2)/p34(cdc2)复合物,但环己酰亚胺抑制了卵细胞质诱导生发泡破裂(GVBD)的能力。然而,用组成型活性丝裂原活化蛋白激酶(MAPK)激酶2(MEK2)、p33(Ringo)或Delta 90细胞周期蛋白B注射后,经环己酰亚胺处理的卵母细胞发生了GVBD。MAPK活性在第一次减数分裂前刚刚增加,此后保持稳定。虽然注射组成型活性MEK2可诱导GVBD,但用MEK抑制剂U0126或炭疽致死因子处理会延迟GVBD并阻止纺锤体形成。有趣的是,卵细胞质诱导GVBD的能力不受MEK活性抑制的影响。我们的结果表明,为了使卵细胞质在热带爪蟾卵母细胞中诱导GVBD,需要合成一种以不依赖MEK的方式起作用的新的或寿命短的蛋白质。

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