| Summary: | Ubiquitin ligase (E3) plays a versatile role in gonadal development and spermatogenesis in mammals, while its function in fish is little reported. In this study, a Z-chromosome linked ubiquitin ligase <i>rchy1</i> in <i>C. semilaevis</i> (<i>Cs-rchy1</i>) was cloned and characterized. The full-length cDNA was composed of 1962 bp, including 551 bp 5′UTR, 736 bp 3′UTR, and 675 bp ORF encoding a 224-amino-acid (aa) protein. <i>Cs-rchy1</i> was examined among seven different tissues and found to be predominantly expressed in gonads. In testis, <i>Cs-rchy1</i> could be detected from 40 days post hatching (dph) until 3 years post hatching (yph), but there was a significant increase at 6 months post hatching (mph). In comparison, the expression levels in ovary were rather stable among different developmental stages. In situ hybridization showed that <i>Cs-rchy1</i> was mainly localized in germ cells, that is, spermatid and spermatozoa in testis and stage I, II and III oocytes in ovary. In vitro RNA interference found that <i>Cs-rchy1</i> knockdown resulted in the decline of <i>sox9</i> and <i>igf1</i> in ovarian cell line and down-regulation of <i>cyp19a</i> in the testicular cell line. These data suggested that <i>Cs-rchy1</i> might participate in gonadal differentiation and gametogenesis, via regulating steroid hormone synthesis.
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