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Studies on control mechanisms of cytoskeletal networks maintaining the morphology of mammalian eggs

维持哺乳动物卵形态的细胞骨架网络控制机制研究

基本信息

批准号：
17580243
负责人：
SUZUKI Hiroyuki
金额：
$ 1.84万
依托单位：
Hirosaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17580243/
关键词：
Mammals Oocytes Cytoskeleton Microtubules Microfilaments Intermediate filaments Rho kinase MAPK 胚ミトコンドリア

项目摘要

1. Functional roles of cell-cycle control kinase on the keratin distribution of the mammal eggs Influences of MPF, MAPK and Rho kinase on the distribution of keratin in the hamster eggs were investigated by the fluorescent-imaging. The present results clarified that hamster eggs included keratin in the cytoplasm, of which distribution changed dynamically from the concentrated pattern at the GV stage into the fibrous pattern at the M II stage during oocyte maturation. When MPF and MAP kinase were inhibited, the distributional change of the keratin had not happened. On the other hand, no remarkable changes was seen when Rho kinase was inhibited. It is suggested that the distribution of keratin in the mammal eggs may be controlled by MPF and MAP kinase, but not by Rho kinase.2. Functional role of Rho kinase in maturation process of the pig oocytes Functional role of Rho kinase during oocyte maturation was studied in the pig. The present study revealed that the germinal-vesicle (GV) breakdown and expulsion of the first polar body was restrained when Rho kinase was inhibited. However, the separation of the chromosome wasn't affected. In addition, the GV breakdown had occurred even if Rho kinase was blocked 18 hours after maturation incubation. It is suggested that Rho kinase may play a role in the control mechanism which causes GV breakdown by 18 hours of cultivation. Fluorescent observations revealed that the microfilament in the oocytes declined remarkably in density by Rho kinase inhibitor, thereby the first polar body was not released from the oocytes. The oocytes-cumulus cells complexes incubated with Rho kinase inhibitor didn't show the cumulus cell expansion after 44 hr of maturation. The possibility affecting the distribution of microtubules among the cumulus-cells was suggested in addition to the architecture of microfilaments.

1.细胞周期调控激酶在哺乳动物卵细胞角蛋白分布中的作用用荧光成像技术研究了MPF、MAPK和Rho激酶对金黄地鼠卵细胞角蛋白分布的影响。本研究结果表明，金黄地鼠卵在卵母细胞成熟过程中，细胞质中含有角蛋白，其分布由GV期的集中型向M Ⅱ期的纤维型动态变化。当MPF和MAP激酶被抑制时，角蛋白的分布没有发生变化。另一方面，当Rho激酶被抑制时，没有观察到显著的变化。提示角蛋白在哺乳动物卵子中的分布可能受MPF和MAP激酶控制，而不受Rho激酶控制. Rho激酶在猪卵母细胞成熟过程中的作用研究了Rho激酶在猪卵母细胞成熟过程中的作用。本研究表明，抑制Rho激酶可抑制胚泡（GV）的破裂和第一极体的排出。但是，染色体的分离不受影响。此外，即使Rho激酶在成熟孵育后18小时被阻断，GV也发生了分解。这表明Rho激酶可能在导致GV在培养18小时时破裂的控制机制中起作用。荧光观察显示，Rho激酶抑制剂使卵母细胞内微丝密度显著下降，从而导致第一极体不能从卵母细胞中释放出来。卵母细胞-卵丘细胞复合体经Rho激酶抑制剂孵育44小时后，卵丘细胞不再扩增。除了微丝的结构外，还可能影响微管在卵丘细胞中的分布。