斑马鱼卵母细胞第一次减数分裂过程中MAPK的表达及定位

The activity and localization of MAPK during the first meiosis in zebrafish oocytes

用不同浓度的促滤泡激素(FSH)和17α,20β-双羟孕酮分别作用于斑马鱼卵母细胞。结果表明,1 IU/mL的FSH和5 ng/mL的17α,20β-双羟孕酮对斑马鱼卵母细胞生发泡破裂都具有明显促进作用。其中17α,20β-双羟孕酮的作用效果较好,经过SDS-PDGE电泳和WesternBlot检测发现,5 ng/mL 17α,20β-双羟孕酮对斑马鱼卵母细胞诱导后,分裂原蛋白激酶被激活,生发泡破裂的卵母细胞数目增多。免疫组化检测证实,第Ⅰ期的卵母细胞中分裂原蛋白激酶没有激活,第Ⅱ期、第Ⅲ期细胞质中分裂原蛋白激酶开始激活,第Ⅴ期卵母细胞的胞质与核区同时存在活性的分裂原蛋白激酶,并伴有生发泡破裂。以上结果说明分裂原蛋白激酶在斑马鱼卵母细胞第一次减数分裂恢复过程中发挥重要作用,并且随着分裂原蛋白激酶激活,其活性部位从细胞质转移到细胞核。

Zebrafish oocyte maturation was induced by various concentrations of FSH and 17α,20β-DHP.The results indicated that FSH of 1IU/mL and 17α,20β-DHP of 5ng/mL could initiate significantly germinal vesicle breakdown（GVBD） of zebrafish,separately,while the 17α,20β-DHP of 5ng/mL had better performance.The analysis of SDS-PDGE and Western blot showed that 17α,20β-DHP of 5ng/mL activated oocyte MAPK and increased GVBD in the zebrafish.The histochemistry assay showed MAPK was activated at the stages Ⅱ and Ⅲ of oocyte development in the plasm while no MAPK activity was observed at the stage Ⅰ.At the stage Ⅴ,the activated MAPK could be observed both in the plasm and nucleus while GVBD took place.The results indicated that activated MAPK played an important role in promoting the ovary development and began to move from the plasm to the nucleus while it was activated.