期刊
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
卷 41, 期 4, 页码 728-737出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.dci.2013.08.004
关键词
Caspase-8; Apoptosis; Cadmium; Heavy metal; Common carp
资金
- National Natural Science Foundation of China [31001128]
- Natural Science Foundation of Jiangxi Province [2010GQY0209]
- Technology Pedestal and Society Development Project of Jiangxi Province [2010BSB03001]
- Educational Department of Jiangxi Province [GJJ11053]
Caspase-8, the essential initiator caspase, is believed to play a pivotal role in death receptor-mediated apoptotic pathway. It also participates in mitochondria-mediated apoptosis via cleavage of proapoptotic Bid in mammals. However, its role in fish remains elusive in Cadmium-induced apoptotic pathway. In this study, we isolated the caspase-8 gene from common carp, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of caspase-8 comprised 475 amino acids, which showed approximately 64.1% identity and 79.8% similarity to zebrafish (Danio rerio) caspase-8, possessed two conserved death effector domains, a large subunit and a small subunit. Phylogenetic analysis demonstrated that caspase-8 formed a clade with zebrafish caspase-8. In kidney, cadmium (Cd) exposure triggered apoptosis and increased caspase-3 and -9 activities, whereas it did not affect caspase-8 activity. Real-time quantitative PCR analysis revealed that caspase-8 transcriptional level was not significantly increased in kidney after exposure to Cd. Using Western blot analysis, no caspase-8 cleaved fragment was detected and no significant alteration of procaspase-8 level was found with the same Cd-treated condition. Moreover, the immunopositive staining was predominantly limited to the cytoplasm of renal tubular epithelial cells and no remarkable changes of immunoreactivities were observed using immunohistochemical detection after Cd treatment. The results reveal that Cd can trigger apoptoSis, while it cannot activate caspase-8 in purse red common carp. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.
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