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目的 通过观察精子膜蛋白Cyritestin单抗对体外鼠精 卵结合和融合作用的影响 ,探讨该蛋白在受精过程中的作用。方法 应用鼠Cyritestin单抗 ,通过体外受精试验观察该单抗预孵育不同浓度精子后 ,对精 卵结合和融合功能的影响 ;应用免疫细胞化学方法确定Cyritestin在精子表面的特异性定位。结果 不同精子浓度下 ,Cyritestin单抗均可显著降低体外精 卵的结合和融合能力 ;可使精 卵结合指数降低 3 7.18%～ 79.0 0 % ,平均 60 .3 3 % ;使精 卵融合的受精指数降低 5 1.66%～ 77.0 1% ,平均 66.90 % ;受精率下降3 2 .2 8%～ 76.15 % ,平均 5 7.41%。在精子顶体反应前后 ,Cyritestin在精子表面均特异性定位于精子头部赤道区域和顶体内膜区域。结论 Cyritestin在精 卵的结合和融合中均发挥重要生理作用。该蛋白作为精子靶抗原开发避孕疫苗的可能性值得深入研究。
Objective To investigate the effect of the sperm membrane protein Cyritestin monoclonal antibody on the binding and fusion of murine sperm and egg in vitro and to explore its role in the process of fertilization. Methods The mouse Cyritestin monoclonal antibody was used to observe the effect of Cyto-MS on the sperm-egg binding and fusion function after pre-incubation with different concentrations of sperm in vitro fertilization test. Cyritestin specific localization on sperm surface was determined by immunocytochemistry. Results Cyritestin monoclonal antibody could significantly reduce the binding and fusion ability of sperm in vitro under different sperm concentrations. The sperm-egg combination index could be reduced by 31.78% -79.0 0% with an average of 60.33% Index decreased 5 1.66% ~ 77.0 1%, an average of 66.90%; fertilization rate decreased 32.28% ~ 76.15%, an average of 5 7.41%. Before and after sperm acrosome reaction, Cyritestin is located on the sperm surface both in the equatorial region of sperm head and in the acrosome region. Conclusion Cyritestin plays an important physiological role in sperm-egg binding and fusion. The possibility of developing this contraceptive vaccine as a sperm target antigen deserves further study.