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Objective Hundreds of HSP40s derived from various species have been identified, of which several proteins are involved in spermatogenesis.DNAJB13 is a type Ⅱ HSP40/ DnaJ protein.In a previous study, we cloned mouse Dnajb13, which is up-regulated in cryptorchidism.To date, however, little is known about the localization and functions of DNAJB13 during spermatogenesis.This study intends to identify the expression pattern of DNAJB13 during mammalian spermatogenesis.Methods We performed RT-PCR and western blot to study the expression profile of Dnajb13 mRNA and DNAJB 13 protein in different tissues of adult BALB/c mice.And testes from different developmental periods of BALB/c mice were collected for quantitative PCR, western blot, in situ hybridization and immunohistochemical detection to study the expression of DNAJB13 protein in mouse spermatogenesis.Results RT-PCR and western blot revealed that the Dnajb13 gene and DNAJB13 protein were expressed not only in the testis but also in several other ciliated cell-containing tissues like the trachea, lung and oviduct.Quantitative PCR showed that the expression of Dnajb13 mRNA in testis was detectable as early as postnatal week 1, and sharply increased from postnatal week 3.Western blotting and immunohistochemistry determined that the DNAJB 13 protein, which was located in the cytoplasm of spermatids and the sperm flagellum, was detectable from postnatal week 4.Conclusions Based on the spatiotemporal expression observed in the cytoplasm of spermatids and sperm flagella, we suggest that DNAJB13 participates in spermiogenesis and the motility of mature spermatozoa.