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。
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-ERp27 group (co-incubated with ERp27 antibody). 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 II oocytes. The fertilization rate (90.91% vs. 89.13% vs. 76.00%) was not significantly different among the blank control, negative control and anti ERp27 groups (P〉0.05). The fertilization index of anti ERp27 group (3. 20± 0. 92) was significantly lower than the other two groups [(5. 20±1. 32) , (5. 40±1.07) ](P〈0. 05). Conclusions: ERp27 was expressed in MI and M Ⅱ stage oocytes in mouse and this protein may play a role in facilitating sperm-oocyte fusion.