论文部分内容阅读
目的:利用特异性小发夹RNA(shRNA)表达质粒,研究IA型磷脂酰肌醇3-激酶(PI3Ks)催化亚基p110α对小鼠受精卵发育的影响。方法:把p110αshRNA表达质粒显微注射到受精卵中。确定能干扰p110α表达的shRNA的最适浓度。通过检测RNA干扰后的p110αmRNA水平和蛋白表达情况,证明干扰是否成功。在确定成功干扰p110α后,观察受精卵的形态变化、分裂率以及对MPF活性的影响。结果:p110α表达受到干扰后,MPF的活性明显降低,受精卵第一次有丝分裂受到干扰,卵裂率明显降低。结论:p110α可能通过影响MPF的活性,调控小鼠受精卵的早期发育,为进一步探讨PI3K信号转导通路在小鼠胚胎中的作用奠定了基础。
OBJECTIVE: To study the effects of type I phosphatidylinositol 3-kinase (PI3Ks) catalytic subunit p110α on the development of mouse fertilized eggs by using specific shRNA expression plasmids. Methods: The p110α shRNA expression plasmid was microinjected into zygotes. The optimal concentration of shRNA that can interfere with p110α expression was determined. By detecting the RNA interference p110αmRNA levels and protein expression, to prove the interference was successful. After confirming the successful interference of p110α, morphological changes, cleavage rates and effects on MPF activity were observed. Results: After the expression of p110α was disturbed, the activity of MPF was significantly reduced. The first mitosis of fertilized eggs was disturbed and the cleavage rate was significantly decreased. Conclusion: p110α may regulate the early development of mouse fertilized eggs by affecting the activity of MPF, which lays the foundation for further exploring the role of PI3K signal transduction pathway in mouse embryos.