Ddx1基因在不同发育阶段小鼠睾丸组织中的表达

Developmental expression of ddx1 in mouse testis

目的系统观察不同日龄小鼠睾丸组织的发育特点,检测ddx1基因与蛋白在不同发育阶段小鼠睾丸组织中的表达模式,探讨ddx1基因在精子发生过程中的作用。方法采用石蜡切片HE染色的方法观察不同发育阶段小鼠睾丸组织的结构特征;采用实时荧光定量PCR法检测ddx1mRNA在不同日龄小鼠睾丸组织中的表达;采用免疫印迹和免疫组织化学法检测DDX1蛋白在不同日龄小鼠睾丸组织中的表达特性与细胞内的定位。结果石蜡切片HE染色显示5、15、23、35、42和60日龄小鼠睾丸组织结构特点能代表生精上皮组织发生发育阶段;实时荧光定量PCR法显示ddx1mRNA在各日龄小鼠睾丸组织中均有表达,于15日龄开始增高,随后维持稳定水平;免疫印迹法显示DDX1蛋白在35、42、60日龄小鼠睾丸组织中特异性表达并逐渐增高;免疫组织化学法显示在少量精原细胞、大量精母细胞和圆形精子细胞胞质与细胞核中存在DDX1蛋白阳性表达。结论 Ddx1基因和蛋白在小鼠精子发生过程中呈现阶段与细胞特异性表达,提示ddx1基因可能在精子发生过程中发挥作用。

Objective To identify the different developmental stages of testis in mice.To investigate the dynamic expression of ddx1 mRNA and protein in mouse testis as well as the function of ddx1 during spermatogenesis.Methods Paraffin sections were used with HE staining to show morphological structure of normal mouse spermatogenesis at postnatal stages of the developing testis.Quantitative real-time PCR was used to establish the ddx1 mRNA expression in the testis of mice at different ages.Immunohistochemistry and Western blotting were carried out to detect the expression and distribution of DDX1 protein in mouse testis and spermatocytes.Results Histological analysis showed morphological structure of normal mouse spermatogenesis at different postnatal stages of the developing testis from 5,15,23,35,42,60-day old mice.Quantitative real-time PCR showed that express of ddx1 mRNA situate at low relatively at postnatal P5,then increased at P15 and retained high afterwards.Western blotting confirmed that the specific expression of DDX1 protein in mouse testis.The DDX1 antigen was detected in testis of 35,42 and 60-day old and located mainly in the cytoplasm and nuclear of spermatocyte and round spermatid by immunohistochemistry.Conclusion The ddx1 mRNA and protein expressed in the testis of mice at different stage during spermatogenesis.It indicated that ddx1 may be involved in the regulation of spermatogenesis and spermiogenesis.