Objective To analyze the expression characteristics of SRPK2 (serine/arginine-rich protein specific kinase 2 mRNA and its encoded protein products in mouse testis, and to reveal its molecular role during spermatogenesis. Methods Using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting to analyze the expression of SRPK2 mRNA and its protein products in several mouse tissues. The stage-specific expression pattern of SRPK2 mRNA in mouse testis was examined by real-time quantitative PCR. Immunohistochemical staining was applied to identify the SRPK2 protein distribution in mouse testicular seminiferous tubules and indirect immunofluorescence staining was used to detect the subcellular localization of SRPK2 protein in spermatic cells. Results Semi-quantitative RT-PCR and Western blotting analysis revealed that SRPK2 was highly expressed in mice testis. Real-time quantitative PCR analysis showed that SRPK2 mRNA was expressed abundantly at the stage of 5-and 8-week old mouse testes, suggesting that SRPK2 gene has stage-specific expression patterns during mouse spermatogenesis. Immunohistochemical and indirect immunofluorescence staining demonstrated that SRPK2 protein was located around the surface of elongated sperm nucleus. Conclusions SRPK2 is highly expressed in mouse testis and has significant stage-specific expression characteristics with distinct nuclear localization. These results indicate that SRPK2 may participate in precursor mRNA splicing during mouse spermiogenesis. The molecular mechanism of SRPK2 is yet to be further studied.