Studies On The Expression Pattern And Function Of Ezrin In Mouse Ovaries

Ezrin has been proved to have relation with mouse embryonic development and embryo implantation in rat uterine. However, there were few documents about ezrin and Phosphorylation ezrin effect on the expression pattern of mouse ovary and the regulation of the development of mouse oocytes. Here we investigated ezrin on the expression pattern of mouse ovary, the follicles of mouse and the regulation of the development of mouse oocytes. There were two parts in this study: 1.The expression of Ezrin on different days of mouse ovaries and different developmental stages of mouse follicles.By quantitative PCR and immunohistochemical methods to detect Ezrin mRNA transcription and Ezrin protein in different days of mouse ovaries. Results showed that:Ezrin mRNA of mouse ovaries were transcribed in different days, the lowest was newborn 10d mouse ovaries, the maximum 10w sexual mouse ovaries mature. The expression pattern was with the greater mouse-week-old, showed increased expression of the trend.Ezrin protein were expressed in mouse ovaries of different days by immunohistochemical staining, the highest expression of the mature ovary 10w mouse. Ezrin mRNA transcripted levels of PMSG treatment groups were higher than normal mouse ovaries. The highest expression were PMSG treatment group 6w. By immunohistochemical staining,PMSG treatment group 6w oocytes were deeperr colored, PMSG treatment group 10w of stromal cells were deeper than other groups.Simultaneously the normal and superovulation of mouse ovary by immunohistochemical staining to detect phosphorylation Ezrin protein; Results showed that phosphorylation Ezrin protein were expressed in mouse ovaries of different days.At the same time, we detected different developmental stages of follicles in mice by quantitative PCR:Ezrin mRNA transcripts were found in preantral follicles and antral follicles, the antral follicles were significantly higher than preantral follicles. The difference was significant (P<0.05). 2. Ezrin were Expressed at different developmental stages of mouse oocytesBy quantitative PCR methods and indirect immunofluorescence technique showed that:Ezrin mRNA transcription observed GV oocytes, MⅡ oocytes and MⅡ oocytes in vitro maturation. GV of the lowest, in vivo MⅡ of the highest,in vitro matured MⅡ oocytes was significantly lower than in vivo matured oocytes, the Ezrin mRNA transcription of various developmental stages were significantly different (P <0.05). Immunofluorescence staining showed that Ezrin protein and phosphorylation Ezrin protein were expressed at different developmental stages in mouse oocytes, the highest expression levels in vivo MⅡ oocytes.We injected Ezrin siRNA and negative controls by microinjection to GV oocytes and cultured in vitro 3h, 10h,18h, counted the oocyte maturation rate, as the first polar body of mature standards. The results showed that the maturation rate of Ezrin siRNA injection group was significantly lower than the negative control group. Also collected stagnant GV oocytes and MⅡ oocytes, Ezrin expression and spindle positioning was detected by indirect immunofluorescence. The results showed that the fluorescence intensity of Ezrin siRNA injection group was lower than the control group, the position of the spindle showed abnormal positioning compared with the control group.