小鼠睾丸曲细精管生精上皮的分离及形态观察

Separation and Morphological Observation of Mouse Testis Seminiferous Tubule Epithelium

目的寻找一种简单可行的研究精子发生过程中蛋白质定位的方法。方法将新鲜的小鼠睾丸去除白膜,通过胶原酶Ⅰ及DNA酶Ⅰ的作用分离各种细胞。细胞经简单处理后以合适的浓度铺片,进行荧光染色,并于共聚焦荧光显微镜下观察。结果通过分析特异的染料对顶体和细胞核染色的结果,可以分辨出各个阶段的生精细胞。结论建立了一种研究精子发生过程中蛋白质定位的简单、可靠的方法。

Objective To explore a simple and feasible technique to locate proteins during spermatogenesis.Methods Various germ cells and somatic cells were separated by collagenase I and DNase I after the albuginea was removed.The cells were then smeared,dyed,and observed directly under fluorescence and confocal microscopy.Results Germ cells at different steps were successfully identified by specific dyestuffs for acrosome and nucleolus.Conclusion A simple method for locating proteins during permatogenesis was successfully developed.