【摘要】 目的探讨内质网蛋白27(ERp27)在小鼠卵母细胞的表达定位,以及该蛋白在小鼠精卵融合中的作用。方法将性成熟后的小鼠促排卵后,取出卵母细胞,并分为空白对照组(不做任何处理)、阴性对照组(与正常血清IgG共孵育)、抗ERp27抗体封闭组(与抗ERp27单克隆抗体共孵育),用免疫荧光法检测ERp27在小鼠卵母细胞上的定位;通过穿卵实验观察抗ERp27抗体对受精率、受精指数的影响。结果免疫荧光结果显示ERp27在小鼠MI期卵母细胞主要分布在胞浆的周边区域中,在MⅡ期卵母细胞则均匀分布于卵母细胞胞质;穿卵实验结果显示,空白对照组、阴性对照组及抗ERp27抗体封闭组受精率分别为90.91%、89.13%和76.00%,差异无统计学意义(P>0.05),受精指数分别为(5.20±1.32)、(5.40±1.07)和(3.20±0.92),抗ERp27抗体封闭组的受精指数显著低于空白对照组和阴性对照组,差异有统计学意义(P<0.05)。结论 ERp27在小鼠MⅠ及MⅡ期卵母细胞中均有表达,该蛋白可能与精卵融合有关。更多还原

【Abstract】 Objective:To investigate the localization of endoplasmic reticulum protein 27(ERp27)on mouse oocytes and its role in gamete fusion.Methods: The oocytes were collected from sexually mature male mice after controlled ovarian hyperstimulation.The oocytes were randomly divided into blank control(no treatment)group,negative control(co-incubated with IgG)group and anti-ERp27group(co-incubated with ERp27antibody).The localization of ERp27 was detected by immunofluorescence analysis.The effect of anti-ERp27 antibody on fertilization rate(FR)and fertilization index(FI)was observed by sperm-oocyte fusion assay.Results:According to indirect immunofluorescence analysis,ERp27 was mainly distributed in the cytoplasm of the peripheral region in M Ⅰ oocytes,while evenly distributed in the cytoplasm in M Ⅱoocytes.The fertilization rate(90.91% vs.89.13% vs.76.00%)was not significantly different among the blank control,negative control and anti-ERp27groups(P>0.05).The fertilization index of antiERp27group(3.20±0.92)was significantly lower than the other two groups [(5.20±1.32),(5.40±1.07)](P<0.05).Conclusions:ERp27was expressed in MI and MⅡstage oocytes in mouse and this protein may play a role in facilitating sperm-oocyte fusion.更多还原