Molecular Characterization and Functional Study of Insulin-Like Androgenic Gland Hormone Gene in the Red Swamp Crayfish, .

The androgenic gland (AG) is a male-specific endocrine organ that controls the primary and secondary sexual characteristics in male crustaceans. More evidence indicates that the insulin-like androgenic gland hormone gene () is the key male sexual differentiation factor, particularly the application of RNA interference (RNAi) technology on . In this study, the full-length cDNA of (termed ) was isolated from the red swamp crayfish, . Tissue distribution analysis showed that in addition to its expression in the AG of male , was widely expressed in female tissues and other male tissues. The PcIAG protein was detected in the reproductive and nervous systems of adult male . Additionally, RNAi results showed that the expression could be silenced efficiently, and the male sperm maturation and release possibly present a transient adverse interference at lower doses (0.1 μg/g and 1 μg/g) of PcIAG-dsRNA ( double-stranded RNA). Dramatically, the expression level of increased sharply shortly after the injection of higher doses (5 μg/g and 10 μg/g) of PcIAG-dsRNA, which might accelerate the maturation and release of sperm. Moreover, the expression of () was detected by Quantitative Real-Time PCR (qPCR) after the injection of PcIAG-dsRNA to explore whether the gene regulates the gene, and we found that the did not directly regulate in . The study could help accelerate the progress of functional research and provide a useful reference for the single-sex selective breeding of .