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目的 研究中心体蛋白centrin在大鼠生精细胞中的表达情况 ,以深入了解centrin在精子发生过程中的作用。 方法 通过重力沉降法分离大鼠不同发育阶段的生精细胞 ,用免疫荧光和蛋白印迹实验检测各级生精细胞中centrin蛋白的表达 ,用定量RT PCR检测centrin同源基因centrin1和centrin2mRNA的表达水平。 结果 间接免疫荧光和蛋白印迹显示精母细胞、圆形、长形精子细胞均有centrin蛋白存在 ,位于中心粒上 ,而在附睾的成熟精子中centrin则消失。RT PCR研究发现 ,centrin1在睾丸组织中特异性表达 ,centrin2在多种组织中均有表达。在睾丸中 ,centrin1仅在生精细胞进入减数分裂后转录 ,其mRNA水平在圆形精子细胞中最高 ,而centrin2在精原细胞中即有表达 ,减数分裂后其mRNA难以检测到。 结论 centrin蛋白在大鼠雄性配子的发育过程中最终丢失 ;该基因家族中同源基因centrin1和centrin2表达呈现组织特异性和发育阶段特异性 ,在精子发生过程中发挥不同功能 ,centrin1蛋白可能与减数分裂及鞭毛生成相关 ,centrin2则参与细胞有丝分裂过程
Objective To study the expression of central centrin protein in rat spermatogenic cells in order to further understand the role of centrin in spermatogenesis. Methods The spermatogenic cells in different developmental stages of rats were isolated by gravity sedimentation. The centrin expression in spermatogenic cells was detected by immunofluorescence and Western blotting. The expression of centrin1 and centrin2 mRNA was detected by quantitative RT-PCR. . Results Indirect immunofluorescence and Western blotting showed that centrin was present in spermatocytes, round and elongated spermatids, located on the centrioles, whereas centrin disappeared in mature spermatozoa of epididymis. RT PCR study found that centrin1 is specifically expressed in testicular tissue, centrin2 is expressed in many tissues. In testis, centrin1 transcribes only after spermatogenic cells enter meiosis, and its mRNA level is highest in round spermatids, whereas centrin2 is expressed in spermatogonia, and its mRNA is difficult to detect after meiosis. CONCLUSION centrin protein is lost during the development of male gametes in rats. The expression of centrin1 and centrin2 genes in this gene family is tissue-specific and developmental-specific and exerts different functions during spermatogenesis. Mitosis and the formation of flagella, centrin2 is involved in cell mitosis