cAMP/PKA诱导卵母细胞减数分裂停滞的机制

大多数物种的卵母细胞在减数分裂前都要经历长时间停滞,其中cAMP对卵母细胞减数分裂停滞具有重要作用,本研究关注c AMP对卵母细胞减数分裂的影响及其机制。本研究通过将卵母细胞与cAMP预孵育,再用胰岛素刺激研究胰岛素诱导的卵母细胞成熟的影响,接着本研究通过显微注射和Zeiss 100TV显微镜分析cAMP对PKA在卵母细胞中定位的影响,并且本研究用Western blotting的方法研究cAMP/PKA对mos蛋白的表达和MAPK蛋白磷酸化的影响。结果显示,本研究通过亲和层析得到了高纯度的PKA蛋白,且cAMP/PKA能够抑制卵母细胞的成熟,而PKA的热稳定抑制剂PKI能够解除PKA对卵母细胞减数分裂的抑制,cAMP/PKA也能够影响mos的积累以及MAPK的磷酸化。cAMP能够影响PKA在卵母细胞中的定位,cAMP/PKA能够通过影响mos积累抑制卵母细胞的减数分裂,这可能与cAMP能够抑制MAPK磷酸化有关。

Mechanism of cAMP/PKA-induced Meiosis Arrest in Oocytes

Oocytes of most species experience long periods of stagnation before meiosis,and cAMP plays an important role in oocyte meiosis stagnation.This study focused on the effect of cAMP on oocyte meiosis and its mechanism.In this study,oocytes were pre-incubated with camp,then insulin stimulation was used to study the effects of insulin-induced maturation of oocytes,then the effect of cAMP on the localization of PKA in oocytes was analyzed by microinjection and Zeiss 100 TV microscope,and the effect of cAMP/PKA on the expression of mos protein and the phosphorylation of MAPK protein was studied by Western blotting.The results showed that high purity PKA protein was obtained by affinity chromatography in this study,and cAMP/PKA could inhibit oocyte maturation,while PKI,a thermal stability inhibitor of PKA,could relieve the inhibition of PKA on meiosis of oocytes,and cAMP/PKA could also affect the accumulation of MOS and the phosphorylation of MAPK.CAMP can affect the location of PKA in oocytes,and cAMP/PKA can inhibit the meiosis of oocytes by affecting MOS accumulation,which may be related to the inhibition of MAPK phosphorylation by cAMP.