Three representative subtypes of caspase in miiuy croaker: Genomic organization, evolution and immune responses to bacterial challenge

Caspase proteins are intracellular proteases which function as initiators and effectors of apoptosis. According their difference of functions, the caspases can be divided into apoptosis related caspases and inflammatory mediator, and the former included apoptosis activator and apoptosis executioner. In this study, three different subtype caspases (caspase1, caspase3 and caspase9) from Miichthys miiuy miiuy croaker were analyzed. The caspase1 belongs to the inflammatory mediator, caspase3 belongs to apoptosis executioner, and caspase9 belongs to apoptosis activator. Miichthys miiuy caspase1 (Mmcaspase1) and Mmcaspase9 exhibited three conserved domains including a CARD, a large subunit p20 and a small subunit p10. The Mmcaspase3 encoded two conserved domains including a large subunit p20 and a small subunit p10. Mmcaspase3 and Mmcaspase9 contained a histidine active sequence and a cysteine active sequence. However, Mmcaspase1 only contained the cysteine active sequence. The real-time PCR (RT-PCR) analysis showed these three caspases were expressed constitutively in all examined tissues in miiuy croaker, although the expression levels varied from tissue to tissue. Expression analysis showed that Mmcaspase1 was up-regulated obviously in liver, spleen and kidney, and indicated its positive role in response to Vibrio anguillarum infection, but Mmcaspase3 and Mmcaspase9 showed different expression pattern in liver, spleen and kidney, its showed that different subtypes of caspase having different immune response mechanisms. These results revealed that the organs adjusted the expressions of these three genes with the infected of pathogens, suggesting the immunoprotection of these genes.