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Background  Caspases are a family of conserved intracellular cysteine-dependent aspartate-specific cysteine proteases that play important roles in regulating cell death and inflammation. Our previous study revealed the importance of the inflammatory  caspase 1  gene in extracellular ATP-mediated immune signaling in Japanese flounder,  Paralichthys olivaceus . To explore the potential roles of other caspases in  P. olivaceus  innate immunity ,  we extended our study by characterizing of the responses of four additional  P. olivaceus caspase  genes ,  termed  JfCaspase 2 ,  3 ,  6  and  8 , to inflammatory challenge and extracellular ATP stimulation.    Results  Sequence analysis revealed that the domain structures of all the Japanese flounder caspase proteins are evolutionarily conserved. Quantitative real-time PCR analysis showed that the  JfCaspase 2 ,  3 ,  6  and  8  genes were expressed ubiquitously but at unequal levels in all examined Japanese flounder normal tissues. In addition, the basal gene expression levels of  JfCaspase 2 ,  3 ,  6  and  8  were higher than those of  Jf C aspase 1  in both Japanese flounder head kidney macrophages (HKMs) and peripheral blood leukocytes (PBLs). Furthermore, immune challenge experiments showed that the inflammatory stimuli LPS and poly(I:C) significantly modulated the expression of the  JfCaspase 2 ,  3 ,  6  and  8  genes in Japanese flounder immune cells. Finally, DNA fragmentation, associated with increased extracellular ATP-induced  JfCaspase 2, 3, 6  and  8  gene expression and enzymatic activity, was inhibited by the caspase inhibitor Z-VAD-FMK in the HKMs.    Conclusion  Our findings demonstrate broad participation of multiple  caspase  genes in response to inflammatory stimulation in Japanese flounder immune cells and provide new evidence for the involvement of caspase(s) in extracellular ATP-induced apoptosis in fish.

bmc veterinary research

Characterization of the responses of the caspase 2, 3, 6 and 8 genes to immune challenges and extracellular ATP stimulation in the Japanese flounder (Paralichthys olivaceus)