蛋白酶体REGγ对雄性小鼠生精功能的影响

Effects of REGγon spermatogenesis of male mouse

目的研究蛋白酶体REGγ对雄性小鼠生精功能的影响。方法利用免疫蛋白印迹检测小鼠睾丸中REGγ的表达;通过组合酶消化法分离精原干细胞,流式细胞仪检测精原干细胞中c-kit及α6-integrin的表达;通过小鼠精子分析仪检测REGγ基因敲除雄鼠的精子浓度和精子活力;进行小鼠合笼实验检测正常雌鼠和REGγ基因敲除雄鼠合笼后的生育力。结果 REGγ在小鼠睾丸中高表达;应用组合酶消化法得到了纯度70%的精原干细胞;REGγ基因敲除雄鼠精原干细胞的c-kit及α6-integrin表达量均显著低于野生型组(P<0.05);REGγ基因敲除型雄鼠的平均精子浓度(37.1×106/ml)和精子活力(54%)均显著低于野生型雄鼠(75.4×106/ml、74%)(P<0.05);REGγ基因敲除型雄鼠与野生型雌鼠合笼后的产仔数均低于野生型雄鼠(P<0.05)。结论蛋白酶体REGγ参与调节雄鼠的生精功能。

Objective： To study the effect of REGγ on mouse spermatogenisis. Methods： The expression of REGγ in mouse testis was detected by using Western blot. The spermatogonial stern cells was isolated by enzymatic digestion method. The expression of c-kit and a6- integrin in spermatogonial stem cells were detected by using flow cytometry. The sperm concentration and motility of REGγ knockout mouse were analyzed by computer assisted sperm analysis system. The fertility of offspring of REGγ knockout and wild type mouse was investigated by mating experiment. Results： REGγ was highly expressed in mouse testis. About 70 % purity of spermatogonial stem cells were obtained by enzymatic digestion method. The expression of c-kit and α6-integrin in spermatogonial stem cells from knockout mice was significant lower than that from wild type mice （P〈 0.05）. Besides, both sperm concentration （37. 1×10^6/ml vs. 75. 4 ×10^6/ml） and sperm motility （54% vs. 74%） in knockout mice were significantly lower inREG7 wild type mice （P〈0. 05）. The numbers of offspring of REGγ knockout of REGγ mice were significantly less than that of wild type male mice （P〈0.05）.Conclusions： REGγ is involved in regulation of spermatogenisis.