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)
Abstract 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-mediat...
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| Format: | Article |
| Language: | English |
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BMC
2019-01-01
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| Series: | BMC Veterinary Research |
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| Online Access: | http://link.springer.com/article/10.1186/s12917-018-1763-y |
| _version_ | 1811344726103687168 |
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| author | Shuo Li Jiafang Li Weijiao Peng Gaixiang Hao Jinsheng Sun |
| author_facet | Shuo Li Jiafang Li Weijiao Peng Gaixiang Hao Jinsheng Sun |
| author_sort | Shuo Li |
| collection | DOAJ |
| description | Abstract 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 JfCaspase 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. |
| first_indexed | 2024-04-13T19:52:24Z |
| format | Article |
| id | doaj.art-f61525c97e7d43e6a3c32e4771c44247 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-13T19:52:24Z |
| publishDate | 2019-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-f61525c97e7d43e6a3c32e4771c442472022-12-22T02:32:29ZengBMCBMC Veterinary Research1746-61482019-01-0115111210.1186/s12917-018-1763-yCharacterization 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)Shuo Li0Jiafang Li1Weijiao Peng2Gaixiang Hao3Jinsheng Sun4Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal UniversityTianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal UniversityTianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal UniversityTianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal UniversityTianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal UniversityAbstract 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 JfCaspase 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.http://link.springer.com/article/10.1186/s12917-018-1763-yExtracellular ATPCaspasesImmune responseApoptosisTeleostParalichthys olivaceus |
| spellingShingle | Shuo Li Jiafang Li Weijiao Peng Gaixiang Hao Jinsheng Sun 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) BMC Veterinary Research Extracellular ATP Caspases Immune response Apoptosis Teleost Paralichthys olivaceus |
| title | 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) |
| title_full | 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) |
| title_fullStr | 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) |
| title_full_unstemmed | 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) |
| title_short | 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) |
| title_sort | 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 |
| topic | Extracellular ATP Caspases Immune response Apoptosis Teleost Paralichthys olivaceus |
| url | http://link.springer.com/article/10.1186/s12917-018-1763-y |
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