Caspase -1, -3, -8 and antioxidant enzyme genes are key molecular effectors following Vibrio parahaemolyticus and Aeromonas veronii infection in fish leukocytes

Caspases are a family of proteases involved in many important biological processes including apoptosis and inflammation. In order to get insights into the caspase gene family and antioxidant enzymes in Totoaba macdonaldi during bacterial infection, an in vitro assay was performed involving three different types of caspases (Casp-1, Casp-3 and Casp-8) and antioxidant enzymes (catalase, gluthathione peroxidase 1 and 4) after Vibrio parahaemolyticus and Aeromonas veronii infection, using head-kidney and spleen leukocytes from the teleost fish totoaba at 12 and 24 h post-exposure. Characterization of caspases by bioinformatics analyses showed that TmCas-1, TmCas-3 and TmCas-8 shared overall sequence identities of 82-61%, 85-97% and 77-63%, respectively, with other teleost fish. Caspase-1, -3 and -8 proteins revealed a conserved penta-peptide sequence at the catalytic site and three amino acid residues involved in the catalysis (H, G and C), as well as two conserved domains. The expression levels of the three caspases were detected in a wide range of fish tissues; however, they varied among tissues and caspases, which were highly up-regulated in immune organs, such as head-kidney, liver and/or spleen. The pathogen-induced gene expression pattern revealed two interesting facts; first, that the expression of all the caspase genes and antioxidant enzyme genes evaluated in this study were strongly induced following V. parahaemolyticus infection; second, these up-regulations reached a maximum level at 24 h post-infection in head-kidney whereas in spleen leukocytes, it was observed at 6-h post-infection. In conclusion, based on these observations, the acute toxic effects of V. parahaemolyticus are associated to cell death and release of free radicals. This information provides a better understanding of the effects and nature of early immune response against common bacterial infections in totoaba leukocytes.