Spermatogenesis is composed of a series of complex biological events, which are regulated by complex factors. There is a phenomenon of delayed translation in spermatogenesis, so the changes of transcription and protein expression are not completely consistent. Thus post-translational modifications (PTMs) play a key role in spermatogenic biological events. In recent years, the development of proteomics has deepened the discovery of PTM. This paper reviews the advances in multiple PTMs proteomic during testicular spermatogenesis. Their effects on sperm function and fertility, as well as their significance for future diagnosis and treatment are discussed.