Ddx3基因在不同日龄小鼠睾丸组织中的表达

Expression of Ddx3 in the testes of mice of different days old

目的:观察小鼠睾丸初次生精过程中的发育特点;检测Ddx3基因与蛋白在小鼠睾丸生精过程中的表达模式;探讨Ddx3基因对小鼠睾丸生精的影响。方法:采用HE染色与PAS染色的方法观察不同日龄小鼠睾丸组织的石蜡切片;用荧光实时定量PCR的方法检测Ddx3 mRNA在小鼠睾丸精子发生过程中的表达;采用免疫组织化学和Western blot方法检测DDX3蛋白在小鼠睾丸初次生精过程中的表达特点和细胞内定位。结果:HE染色结果显示4、13、21、35、42和60日龄是小鼠睾丸发生初次生精过程的特定阶段;荧光实时定量PCR结果显示Ddx3 mRNA在4日龄以后小鼠睾丸组织中均有表达,于35日龄开始明显增高,随后表达水平保持稳定;Western blot显示DDX3蛋白在35、42和60日龄小鼠睾丸生精小管中出现特异性表达并逐渐增高;免疫组织化学法显示DDX3蛋白主要在初级精母细胞和精子细胞的细胞质和细胞核中表达。结论:DDX3蛋白在小鼠初次精子发生过程中呈现特异性表达,提示Ddx3基因可能在精母细胞与精子细胞的发育中发挥作用。

Objective： To investigate developmental characters of the initial spermatogenesis of mice,and to explore the specific expression of Ddx3 mRNA and protein in mouse testis as well as the role of Ddx3 during spermatogenesis. Methods： The testis paraffin sections of the mice of different days old were observed by HE staining and PAS staining. Quantitative RT-PCR was used to detect the Ddx3 mRNA expression during the spermatogenesis of the mice. Immunohistochemistry and Western blot were carried out to assay the distribution and expression of DDX3 protein during the initial spermatogenesis of the mice. Results： HE staining showed that4,13,21,35,42,and 60-day-old were the specific stages of the initial spermatogenesis of the mice. Quantitative RT-PCR indicated that the expression level of Ddx3 mRNA was relativelylow at postnatal 4 days（ P4）,then increased at P35 and retained stable afterwards. Western blotsuggested that the protein expression of DDX3 was detected in testis of 35,42 and 60-day-old and increased gradually. Further,immunohistochemistry showed that DDX3 proteinlocated mainly in the cytoplasm and nuclear of spermatocyte and round spermatid. Conclusions： The DDX3 protein is specifically expressed in the testis during spermatogenesis of the mice of difference days old. It suggests that Ddx3 may be involved in the regulation of spermatogenesis and spermiogenesis.