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首页> 外文期刊>Developmental and Comparative Immunology: Ontogeny, Phylogeny, Aging: The Official Journal of the International Society of Developmental and Comparative Immunology >Cadmium triggers kidney cell apoptosis of purse red common carp (Cyprinus carpio) without caspase-8 activation
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Cadmium triggers kidney cell apoptosis of purse red common carp (Cyprinus carpio) without caspase-8 activation

机译：镉触发荷包红鲤（Cyprinus carpio）的肾细胞凋亡而不激活caspase-8

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摘要

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.
机译：Caspase-8是必需的启动子caspase，据信在死亡受体介导的凋亡途径中起关键作用。它也通过切割哺乳动物中的促凋亡投标而参与线粒体介导的凋亡。但是，它在鱼类中的作用在镉诱导的凋亡途径中仍然难以捉摸。在这项研究中，我们通过快速扩增cDNA末端（RACE）从鲤鱼中分离了caspase-8基因，鲤鱼是中国最重要的工业水生动物之一。推导的caspase-8氨基酸序列包含475个氨基酸，与斑马鱼（。Danio rerio）caspase-8具有大约64.1％的同源性和79.8％的相似性，具有两个保守的死亡效应域，一个大亚基和一个小亚基。系统发育分析表明，caspase-8与斑马鱼caspase-8形成了进化枝。在肾脏中，镉（Cd）的暴露触发细胞凋亡并增加caspase-3和-9活性，而它并不影响caspase-8活性。实时定量PCR分析显示，暴露于Cd后肾脏中的caspase-8转录水平没有显着增加。使用蛋白质印迹分析，在相同的Cd处理条件下，未检测到caspase-8裂解的片段，也未发现procaspase-8水平的显着改变。此外，免疫阳性染色主要限于肾小管上皮细胞的细胞质，Cd处理后使用免疫组织化学检测未观察到明显的免疫反应性变化。结果表明，镉可以触发细胞凋亡，而不能激活荷包红鲤鱼中的胱天蛋白酶8。

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