The Dmrt1 gene: Identification and functional analysis in testicular development and spermatogenesis in the freshwater prawn, Macrobrachium nipponense

Products of the Dmrt gene family are known to harbor a zinc-finger DM domain and play crucial roles as transcription factors in various physiological processes, including sexual differentiation, gonadal development, and spermatogenesis. Despite their significance, research on the Dmrt family in Crustacea remains limited. In the present study, we identified a member of this gene family in the freshwater prawn Macrobrachium nipponense and designated it as MnDmrt1. We investigated its expression patterns across different tissues, potential target genes, and its influence on testicular development and protein interactions within the testes. Our findings revealed that MnDmrt1 encodes a protein composed of 252 amino acids that is predominantly expressed in the testes and localized in spermatogonia, spermatocytes, and spermatozoa. Silencing MnDmrt1 disrupts spermatogenesis and alters the expression levels of several key genes associated with testicular development, spermatogenesis, and sex reversal, such as Dmrt11E, Insulin receptor, and Masculinizer. Additionally, we identified 59 proteins that interact with MnDmrt1 in the testis using pull-down assays and liquid chromatography-tandem mass spectrometry. Notably, the eyestalk negatively regulates the expression level of MnDmrt1 in the testis. Collectively, these results underscore the essential role of MnDmrt1 in testicular development and spermatogenesis in M. nipponense.