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The signaling molecule hedgehog (Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates. Patched 1 (Ptc1), a receptor of Hh, mediates the activity of Hh by binging the Hh ligands. In this study, the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis (CsPtc1) was cloned and characterized. The full-length of CsPtc1 cDNA is 5 212 nucleotides (nt), and encodes a protein with 1 543 amino acid residues. CsPtc1 shared many features with Ptc1 of other telesot species. Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain, liver, heart, gill, intestines, spleen, gonad and kidney, with expression level in testis significantly higher than in ovary. In testis, hybridization signals were mainly detected in primary spermatocytes, secondary spermatocytes, and sertoli cells, while weak signals were found in oocytes. Correspondingly, the degree of methylation is higher in female than in male and pseudomale. Results indicate that CsPtc1 may be involved in Desert Hedgehog (DHH) maintenance of the male and pseudo-male germ line and spermatogenesis.