active substance regulating differentiation of germ lines

1) RT/PCR技术结合Southern blot杂交分析显示，PMSG给药后第1天小鼠卵巢和颗粒细胞FasL mRNA表达水平最高，第5天FasL mRNA表达水平变弱。在小鼠卵巢和经PMSG处理的颗粒细胞中，延长因子（EF-1a） mRNA的组成性表达证明，FasL mRNA的逐渐减少不能归因于闭锁期间细胞RNA的普遍降解。此外，用FasL特异性探针原位杂交分析证实，FasL特异性定位于大多数卵泡颗粒细胞，其表达受PMSG调控。2)我们发现失活的MAPK在GZ期捕获的未成熟卵母细胞中已经存在，并且该失活激酶仅局限于细胞质中。在GZ/M过渡阶段，部分MAPK在生发囊泡破裂（GVBD）之前进入生发囊泡（GV）。此外，免疫印迹分析显示核MAPK以活性形式存在。为了确定这种活性是否能诱导GVBD，我们将活性MAPK微量注射到未成熟的猪卵母细胞中。活性MAPK注入细胞质后迅速失活，不能加速GVBD。相反，MAPK注入GV明显加速GVBD

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