CD9、GPI-AP在卵母细胞表面的表达及其在精卵融合中的作用

目的:探讨CD9(cluster of differentiation9)、CPI-AP(glycosylphosphatidy-linositol anchored proteins)在卵母细胞膜表面的表达及其在精卵融合过程中的作用。方法:用免疫组化法测定CD9和GPI锚定蛋白在正常和未受精卵细胞膜上的表达,通过加入单克隆抗体观察其对受精率的影响,并计算阳性表达率与受精率的关系。结果:①CD9、GPI-AP主要在正常卵母细胞表面表达,在未受精卵母细胞呈低表达,CD9及GPI-AP在正常卵母细胞膜表面的阳性表达率(70.0%,63.3%)明显高于未受精卵母细胞膜表面的阳性表达率(24.0%,28.0%),差异有统计学意义(P0.005)。②随着CD9及GPI-AP单克隆抗体的浓度增加,受精率显著下降,提示CD9在精卵融合中起着重要的作用,且呈剂量依赖性。③受精率与卵母细胞表面CD9、GPI-AP阳性表达率具有相关性(P0.05)。结论:CD9、GPI-AP在精卵融合过程中起重要作用,其检测对精卵融合的发生有预测意义。

Expressions of CD9 and GPI-AP and their functions in sperm-egg fusion

Objective:To explore the expressions of cluster of differentiation 9 (CD9) and glycosylphosphatidy-linositol anchored proteins (GPI-AP) on cell membrane of oocytes and their functions in sperm-egg fusion.Methods:The expressions of CD9 and GPI-AP anchoring proteins on cell membranes of normal cells and unfertilized oocytes were detected by immunohistochemical method,monoclonal antibody was added to observe its effect on fertilization rate,then the relationship between positive expression rate and fertilization rate was calculated.Results:CD9 and GPI-AP mainly expressed on cell membrane of normal oocytes,their expressions on cell membrane of unfertilized oocytes were low,the positive expression rates of CD9 and GPI-AP on cell membrane of normal oocytes were significantly higher than those on cell membrane of unfertilized oocytes (24.0%,28.0%),there was significant difference (P<0.005);the fertilization rate decreased significantly with the increases of CD9 and GPI-AP monoclonal antibody concentrations,which indicated that CD9 played an important role in sperm-egg fusion in a dose-dependent manner;there was positive correlation between fertilization rate and positive expression rates of CD9 and GPI-AP on cell membrane of normal oocytes (P<0.05).Conclusion:CD9 and GPI-AP may play important roles in sperm-egg fusion.The detection of CD9 and GPI-AP can predict sperm-egg fusion.