active substance regulating differentiation of germ lines

调节种系分化的活性物质

基本信息

批准号：
08456147
负责人：
SATO Eimei
金额：
$ 4.48万
依托单位：
Tohoku University (1997)The University of Tokyo (1996)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08456147/
关键词：
fertilization oogenesis spermatogenesis oocytes sperm primordial germ cell embryos in vitro culture

项目摘要

1) RT/PCR technique coupled with Southern blot hybridization analysis showed that highest level of FasL mRNA was demonstrated in murine ovaries and granulosa cells 1 day after the administration of PMSG,while the level of FasL mRNA became very weak on the day 5, respectively. The observed gradual decrease in FasL mRNA could not be attributed to a generalized degradation of cellular RNA during atresia, as evidenced by the presence of constitutive expression of elongation factorla (EF-1a) mRNA in murine ovaries and granulosa cells treated with PMSG.Furthermore, in situ hybridization analysis with a FasL-specific probe confirmed that FasL was specifically localized in the granulosa cells of most follicles and its expression was regulated by PMSG administration.2) We found the the inactive MAPK was already present in immature oocytes arrested at the GZ stage and that this inactive kinase was localized exclusively in the cytoplasm. At the GZ/M transition stage, part of the MAPK moved into the germinal vesicle (GV) before germinal vesicle breakdown (GVBD). In addition, immunoblot analysis showed that the nuclear MAPK existed in an active form. To determine whether this active could induce GVBD,we microinjected active MAPK into immature porcine oocytes. The active MAPK injected into the cytoplasm was quickly inactivated and could not accelerate GVBD.In contrast, MAPK injection into the GV markedly accelerated GVBD

1) RT/PCR技术结合Southern印迹杂交分析显示，在施用PMSG后第1天，小鼠卵巢和颗粒细胞中FasL mRNA的水平最高，而第5天FasL mRNA的水平变得非常弱。。所观察到的 FasL mRNA 的逐渐减少不能归因于闭锁期间细胞 RNA 的普遍降解，正如用 PMSG 处理的小鼠卵巢和颗粒细胞中存在延伸因子 1a (EF-1a) mRNA 的组成型表达所证明的那样。使用 FasL 特异性探针进行的原位杂交分析证实，FasL 特异性定位于大多数卵泡的颗粒细胞中，并且其表达受到以下因素的调节： PMSG 给药。2) 我们发现在 GZ 阶段停滞的未成熟卵母细胞中已经存在失活的 MAPK，并且这种失活的激酶仅位于细胞质中。在 GZ/M 过渡阶段，部分 MAPK 在生发囊泡破裂 (GVBD) 之前移入生发囊泡 (GV)。此外，免疫印迹分析表明核MAPK以活性形式存在。为了确定该活性物质是否可以诱导 GVBD，我们将活性 MAPK 显微注射到未成熟的猪卵母细胞中。注射到细胞质中的活性MAPK很快失活，不能加速GVBD。相反，注射到GV中的MAPK显着加速GVBD