Functional identification and characterization of IpMSTNa, a novel orthologous myostatin (MSTN) gene in channel catfish Ictalurus punctatus

Abstract

Channel catfish (Ictalurus punctatus) are one of the most important commercial freshwater fish in the world. China has been the major producer and consumer of channel catfish following the rapid development in the past three decades. In the present study, a novel orthologous myostatin gene, IpMSTNa, of channel catfish was identified based on homology cloning and genome locating. Multiple sequence alignments and gene structure analyses showed that the IpMSTNa gene and its deduced protein presented similar architectures to other known vertebrates. Phylogenetic and synteny analyses indicated that IpMSTNa belongs to MSTN1 orthologues. Pro-IpMSTNa protein is a typical disulphide-linked homodimer, with each chain containing an N-terminal pro-domain and a C-terminal unmatured GF domain, while pro-IpMSTNa present some significant differences in secondary structure and three-dimensional substances with pro-IpMSTNb. Relative expression level of the IpMSTNa gene upregulated rapidly and decreased dramatically during the embryonic and larval developmental stages, respectively. In addition, IpMSTNa displayed remarkably higher expression at most developmental stages compared to IpMSTNb. Tissue distribution analysis indicated that the IpMSTNa gene had a significantly higher level of expression than IpMSTNb in all selected tissues, with abundantly greater expression in the liver, muscle, gill and spleen, and moderately greater expression in the kidney, intestine, and head kidney. ISH analysis demonstrated that the expression signals of IpMSTNa and IpMSTNb at the selected developmental stages are consistent to qRT-PCR tests. Our study suggested that the IpMSTNa gene may have more biological functions, which have yet to be determined compared to the IpMSTNb gene.