Two spliced variants of insulin-like androgenic gland hormone gene in the Chinese shrimp, Fenneropenaeus chinensis

Abstract

More and more evidence indicates that the insulin-like androgenic gland hormone (IAG) plays an important role in male sexual differentiation in crustaceans. In the present study, two IAG isoforms (Fc-IAG1 and Fc-IAG2) were identified from the penaeid shrimp Fenneropenaeus chinensis. Sequence analysis of IAG gene (Fc-IAG) showed that Fc-IAG1 and Fc-IAG2 were generated by alternative splicing of Fc-IAG pre-mRNA, and they shared almost the same deduced amino acid sequence. Both of them were composed of signal peptide, B chain, C peptide and A chain. They both contained the six conserved cysteine residues and a putative N-linked glycosylated site like IAGs reported in other crustacean species. Tissue distribution and in situ hybridization analysis revealed that they had the highest expression level in the androgenic gland. The transcripts of Fc-IAG1 and Fc-IAG2 could also be detected in hepatopancreas and nerve cord of both sexes at a low expression level. Analysis on their temporal expression profiles showed that they expressed at all embryonic and post-larvae stages. The expression of Fc-IAG1 at different developmental stages displayed a low and stable manner, while the expression of Fc-IAG2 began to increase from post-larvae stages, which suggested that Fc-IAG2 might be involved in male sexual differentiation. In the 5′ flanking sequence of Fc-IAG, putative binding sites for transcription factors regulating transcription of hormone genes and genes related to sexual development were predicted, which provided us a primary understanding on the regulation mechanism of Fc-IAG gene. This is the first time to report the gene structure of IAG gene and distinct variants of IAG transcripts in crustaceans.