Dynamic events are differently mediated by microfilaments, microtubules, and mitogen-activated protein kinase during porcine oocyte maturation and fertilization in vitro
Dynamic events are differently mediated by microfilaments, microtubules, and mitogen-activated protein kinase during porcine oocyte maturation and fertilization in vitro
复制标题
DOI：
10.1095/biolreprod64.3.879
复制
发表时间：
2001-03-01
影响因子：
3.6
通讯作者：
Schatten, H
中科院分区：
文献类型：
--
作者：
Sun, QY;Lai, LX;Schatten, H
微丝、微管和丝裂原活化蛋白（MAP）激酶在猪卵母细胞成熟和受精的几个重要动态事件中的调节作用。荧光显微镜或激光扫描共聚焦显微镜观察荧光标记的微丝、微管和皮质颗粒。丝裂原活化蛋白激酶磷酸化通过Western免疫印迹法显示。我们发现：1）微丝断裂不影响减数分裂的恢复和中期I减数分裂器的形成，但抑制细胞周期的进一步进展（染色体分离）即使MAP激酶被磷酸化; 2）皮质颗粒（CG）的迁移是由微丝驱动的（但不是微管），一旦染色体和CG位于卵膜下，它们在皮层的锚定就不依赖于微丝或微管;（3）卵母细胞激活过程中CG胞吐作用不受微丝和微管的影响：（4）精子的掺入是由微丝介导的，而原核（PN）的融合是由微管而不是微丝控制的; 5）纺锤体微管结构与MAP激酶磷酸化存在时间相关性，而在没有MAP激酶激活的情况下，精子中广泛的微管组织是PN并置和融合所必需的;和6）当微管被诺考达唑破坏或细胞质微管星形细胞被紫杉醇诱导时，MAP激酶磷酸化没有改变。本研究表明，细胞骨架在猪卵母细胞成熟过程中的作用类似于啮齿动物，而猪的受精机制类似于低等脊椎动物。
The role of microfilaments, microtubules, and mitogen-activated protein (MAP) kinase in regulation of several important dynamic events of porcine oocyte maturation and fertilization is described. Fluorescently labeled microfilaments, microtubules, and cortical granules were visualized using either epifluorescence microscopy or laser scanning confocal microscopy. Mitogen-activated protein kinase phosphorylation was revealed by Western immunoblotting. We showed that 1) microfilament disruption did not affect meiosis resumption and metaphase I meiotic apparatus formation but inhibited further cell cycle progression (chromosome separation) even though MAP kinase was phosphorylated; 2) cortical granule (CG) migration was driven by microfilaments (but not microtubules), and once the chromosomes and CGs were localized beneath the oolemma their anchorage to the cortex was independent of either microfilaments or microtubules; 3) neither microfilaments nor microtubules were involved in CG exocytosis during oocyte activation; 4) sperm incorporation was mediated by microfilaments, while pronuclear (PN) syngamy was controlled by microtubules rather than microfilaments; 5) spindle microtubule organization was temporally correlated with MAP kinase phosphorylation, while the extensive microtubule organization in the sperm aster that is required for PN apposition and syngamy occurred in the absence of MAP kinase activation; and 6) MAP kinase phosphorylation did not change either when microtubules were disrupted by nocodazole or when cytoplasmic microtubule asters were induced by taxol. The present study suggests that the role of the cytoskeleton during porcine oocyte maturation is similar to that of rodents, while the mechanisms of fertilization in pig resemble those of lower vertebrates.