关键词: 嘌呤霉素, 精子发生, 精原细胞, 新生蛋白合成

Abstract: Objective Mammalian spermatogenesis is a process with continuous cell proliferation and differentiation composed of a series of spermatogenic cells, and ultimately to produce sperm. However, the proteostasis analysis of spermatogenic cells still lacks effective method. The aim of this study was to explore a method to label and detect nascent protein synthesis in spermatogenic cells in vivo. Method Puromycin ( Puro) , an aminoacyl-tRNA structural analog, was intraperitoneally injected into mice at a dose of 65 mg / kg body weight. 1. 5 hours later, the mice were killed and bilateral testes were isolated. Of these, one of the testes was used for total proteins extraction and the other was used for paraffin sections preparation. Then, the content and localization of Puro in testis were respectively detected by Western blot and immunofluorescence using Puro specific antibody. Result Western blot result showed that Puro could label the overall nascent proteins. Co-immunofluorescence analyses showed that Puro specifically labeled spermatogonia in mouse testis. Conclusion Nascent proteins in mouse spermatogonia can be in vivo labelled by Puro, and their levels can be quantitated through corresponding immunological techniques.

Key words: Puromycin, spermatogenesis, spermatogonia, nascent protein synthesis