Molecular characterization and expression analysis of immunoglobulins (IgM and IgT) heavy chains in black rockfish (Sebastes schlegelii) that response to bacterial challenge

Sebastes schlegelii is a kind of fish with great economic values. Recently, with the continuous expansion of aquaculture scale and the continuous improvement of aquaculture density, outbreak of various diseases has caused huge economic losses to its aquaculture industry. Study of fish immune system can help to understand the mechanism of immune response to external pathogens and can promote the development of immune prevention and control methods. Immunoglobulins (Igs) are complex glycoproteins that appear to be unique to the vertebrates that can recognize a wide variety of pathogens and recruit immune cells and molecules to destroy pathogens, which are generated by a series of rearrangement and somatic mutations. We therefore studied the immunoglobulin genes of S. schlegelii in view of their important roles in resisting to external pathogen infections. In this study, the immunoglobulin heavy chain genes (sIgM, mIgM, sIgT, and mIgT) of S. schlegelii were successfully identified and cloned. Phylogenetic analysis showed that the IgM and IgT genes of S. schlegelii were clustered together with homologous genes of other species, indicating that they were highly conserved during the evolutionary process. Collinearity analysis showed that the immunoglobulin genes and their adjacent genes were aligned with zebrafish, Atlantic salmon and tilapia, which further confirmed the conserved immunoglobulin gene of teleost. Expression analysis of healthy tissues showed that the expression levels of sIgM, sIgT and mIgT were the highest in the skin, while mIgM was the highest in spleen. After different bacterial infection, IgM and IgT were significantly expressed in skin and gill, which may be because skin and gill are the first line of defense against the infection pathogens. Subcellular localization showed that the mIgT protein was expressed in both the cell membrane and cytoplasm. Meanwhile, recombinant protein of mIgT was obtained in vitro, which laid a foundation for subsequent protein function studies. These results provide a theoretical basis for understanding the immunity role of immunoglobulin in S. schlegelii.