| Summary: | <i>Spats1</i> (spermatogenesis-associated, serinerich 1) has been characterized as a male-biased gene which acts an important role in the germ cell differentiation of mammals. Nevertheless, the function of <i>Spats1</i> in the Chinese soft-shelled turtle (<i>P. sinensis</i>) has not yet been reported. To initially explore the expression of <i>Spats1</i> in <i>P. sinensis</i> and its response to sex steroid treatment, we cloned the CDS of <i>Spats1</i> for the first time and analyzed its expression profile in different tissues, including the testes in different seasons. The <i>Spats1</i> cDNA fragment is 1201 base pairs (bp) in length and contains an open reading frame (ORF) of 849 bp, which codes for 283 amino acids. <i>Spats1</i> mRNA was highly expressed in the testes (<i>p</i> < 0.01) and barely detectable in other tissues. In <i>P. sinensis</i>, the relative expression of <i>Spats1</i> also responsive to seasonal changes in testis development. In summer (July) and autumn (October), <i>Spats1</i> gene expression was significantly higher in the testes than in other seasons (<i>p</i> < 0.05). <i>Spats1</i> mRNA was found to be specifically expressed in germ cells by chemical in situ hybridization (CISH), and it was mainly located in primary spermatocytes (Sc1), secondary spermatocytes (Sc2) and spermatozoa (St). <i>Spats1</i> expression in embryos was not significantly changed after 17α-methyltestosterone (MT)and 17β-estradiol (E2) treatment. In adults, MT significantly induced <i>Spats1</i> expression in male <i>P. sinensis</i>. However, the expression of <i>Spats1</i> in testes was not responsive to E2 treatment. In addition, the expression of <i>Spats1</i> in females was not affected by either MT or E2 treatment. These results imply that <i>Spats1</i> is a male-specific expressed gene that is mainly regulated by MT and is closely linked to spermatogenesis and release in <i>P. sinensis.</i>
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