H-Y抗体对小鼠胚胎H-Y抗原表达的研究

Detection of H-Y antigen expression on mouse embryos by H-Y antiboby

BALB/C雄性小鼠脾细胞悬液腹腔注射免疫同系母鼠 ,精子细胞毒性试验筛选抗血清 ,效价高的抗血清分别用于胚胎培养的毒性试验和制备单克隆抗体再辅以间接免疫荧光和PCR对胚胎进行性别鉴别。结果表明 ,直接用H Y抗血清培养的胚胎 ,胚胎退化率达 43 3 % ,与自然性比差异不显著 ;PCR验证间接免疫荧光法鉴定的胚胎性别 ,雄性胚胎准确率为 83 % ,雌性胚胎准确率为 94%。

BALB/C female mice were immunized by intraperitoneal injections of syngeneic male spleen cells and antiserum was detected by sperm cytotoxicity test.Then the chosen antisera were used in mouse embryos culture in vitro and to prepare H-Y monoclonal antibody for later sex detection by indirect immuno-fluorescence assay and polymerase chain reaction(PCR)with amplification of Y chromosome specific sequence.Cultured embryos were classified either affected or unaffected,affected embryos was 43.3% without significant difference from the normal sex ratio.The results from PCR and indirect immuno-fluorescence assay showed that male embryos of H-Y positive was 83%(15/18),female embryos of H-Y negative was 94%(15/16).