Boule基因在小鼠精子发生过程中的动态表达

目的:研究雄性不育相关基因Boule在小鼠出生后首个精子发生波中的动态表达,探讨Boule在精子发生过程中的调控作用。方法:运用免疫印迹法分析BOULE蛋白在多组织中的表达情况。运用实时荧光定量、免疫组织化学和免疫荧光技术检测Boule m RNA和BOULE蛋白在雄性小鼠出生后首个精子发生波各个时间点中的表达。结果:1免疫印迹法显示BOULE蛋白在睾丸中特异性高表达,在出生后16 d睾丸可被检测到,并在随后的发育中睾丸和成年睾丸持续表达;2实时荧光定量方法证实Boule m RNA在雄鼠出生后14 d睾丸中可被检测到,20 d呈现表达高峰;3免疫组织化学显示在精母细胞及圆形精子细胞胞质中存在BOULE蛋白阳性信号。结论:Boule在精子发生过程中的表达是在发育水平精密调控的,在粗线期精母细胞和圆形精子细胞阶段的特异表达提示Boule可能在精母细胞进程和圆形精子细胞中发挥作用。

Dynamic expression of Boule in mouse spermatogenesis

Objective:To study the dynamic expression of male sterility related gene Boule in the first spermatogenesis wave of mouse testes as well as the function of Boule during spermatogenesis. Methods: The expression level of BOULE protein in various tissues and time points were examined using Western blotting. Real-time PCR was used to establish the Boule mRNA expression profile after birth. Immunohistochemistry and immunofluorescence were carried out to analyze the expression and distribution of BOULE protein in mouse testes. Results: Western blot confirmed the specific expression of BOULE protein in mouse testes. Real-time PCR showed that Boule mRNA was first detectable in the testis of mice 20 days after birth. Immunohistochemistry showed that BOULE protein located in the cytoplasm of pachytene spermatocyte and round spermatid during spermatogenesis. Immunofluorescence witnessed the BOULE granule,which formed in the cytoplasm. Conclusion: Boule expression is precisely regulated during spermatogenesis. The specific expression in the cytoplasm of spermatocyte and round spermatid suggested that Boule may be important for spermatocyte progressing and round spermatid differentiation.