跨膜4域亚家族成员6D基因敲除致雄性小鼠生育力降低实验研究

Experimental study of Ms4a6d gene knockout leads to reduced fertility in male mice

目的:探讨跨膜4域亚家族成员6D(Ms4a6d)基因敲除对雄性小鼠生育力的影响。方法:采用聚合酶链式反应(PCR)鉴定小鼠基因型,将Ms4a6d^(-/-)雄鼠和野生型C57小鼠分别与同龄野生型雌鼠合笼交配,统计各组雌鼠怀孕产仔情况,收集睾丸组织,称重并计算器官指数,评估附睾尾精子数量、运动参数等变化情况,通过形态学分析和HE染色评估小鼠睾丸组织形态学变化,采用免疫荧光染色分析睾丸巨噬细胞变化。结果:Ms4a6d^(-/-)雄鼠子代数目显著低于WT雄鼠(P<0.05);Ms4a6d^(-/-)雄鼠睾丸体积及质量均减少(均P<0.05);Ms4a6d^(-/-)雄鼠附睾尾精子浓度下降(P<0.05),各项运动参数未见统计学差异(P>0.05);睾丸组织切片HE染色结果显示Ms4a6d^(-/-)雄鼠睾丸曲细精管直径及管腔厚度均明显减少,支持细胞、精原细胞、初级精母细胞及精子细胞均减少;免疫荧光结果显示Ms4a6d^(-/-)雄鼠睾丸巨噬细胞受到影响。结论:Ms4a6d基因敲除干扰了雄性小鼠睾丸的发育,并造成生精小管直径减少、厚度变薄,附睾尾部精子数量显著下降,从而使成年雄鼠的生育力显著下降。

Objective:To investigate the effect of Ms4a6d gene knockout on the fertility of male mice.Methods:The genotypes of mice were identified by PCR method.Male Ms4a6d^(-/-)male mice and wild type C57 mice were mated in cages with wild type female mice of the same age,respectively.The pregnancy and birth of female mice in each group were counted,the testicular tissue was collected,and the organ index was calculated to evaluate the changes of epididymal tail sperm count and motor parameters.Morphological changes of mouse testicular tissue were evaluated by morphological analysis and HE staining.The changes of testicular macrophages were analyzed by immunofluorescence staining.Results:The number of offspring of Ms4a6d^(-/-)male mice was significantly lower than that of WT male mice,the testicular volume and weight of Ms4a6d^(-/-)male mice decreased,and the epididymal caudal sperm concentration of Ms4a6d^(-/-)male mice decreased(all P<0.05).There was no significant difference in motor parameters(all P>0.05).The results of HE staining of testicular tissue sections showed that the diameter and lumen thickness of testicular convoluted seminiferous tubules,Sertoli cells,spermatogonia,primary spermatocytes and spermatids decreased significantly in Ms4a6d^(-/-)male mice,and immunofluorescence results showed that testicular macrophages in Ms4a6d^(-/-)male mice were affected.Conclusion:Ms4a6d gene knockout interferes with the testicular development of male mice,resulting in a decrease in the diameter and thickness of seminiferous tubules and a significant decrease in the number of sperm in the caudal epididymis,resulting in a significant decrease in fertility in adult male mice.