文章摘要
李坤明,徐文腾,扶晓琴,陈松林.半滑舌鳎蛋白激酶C-alpha(PKCα)的基因克隆和免疫应答分析.渔业科学进展,2020,41(2):69-77
半滑舌鳎蛋白激酶C-alpha(PKCα)的基因克隆和免疫应答分析
Cloning and Immune Response Analysis of Protein Kinase C-alpha (PKCα) in Chinese Tongue Sole (Cynoglossus semilaevis)
投稿时间:2019-02-19  修订日期:2019-03-08
DOI:
中文关键词: 半滑舌鳎  蛋白激酶C-alpha  哈维氏弧菌  免疫应答
英文关键词: Chinese tongue sole Cynoglossus semilaevis  Protein kinase C-alpha  Vibrio harveyi  Immune response
基金项目:
作者单位
李坤明 中国水产科学研究院黄海水产研究所 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071上海海洋大学水产与生命学院 上海 201306 
徐文腾 中国水产科学研究院黄海水产研究所 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071农业农村部海洋渔业可持续发展重点实验室 青岛 266071 
扶晓琴 中国水产科学研究院黄海水产研究所 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071南京农业大学无锡渔业学院 无锡 214081 
陈松林 中国水产科学研究院黄海水产研究所 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071农业农村部海洋渔业可持续发展重点实验室 青岛 266071 
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中文摘要:
      蛋白激酶C(PKC)广泛参与哺乳动物T、B淋巴细胞涉及的免疫反应并发挥重要作用,然而PKC在鱼类免疫过程中的作用却少有报道。本研究在半滑舌鳎(Cynoglossus semilaevis)中克隆了PKC家族的典型成员PKCα(命名为CsPKCα),并进行了分子特征和表达模式分析。CsPKCα的cDNA全长为2315 bp,其中,开放阅读框(ORF)为2013 bp。qRT-PCR分析显示,CsPKCα在各个组织中广泛表达,其中,在鳃、肝脏、皮肤和肠中具有较高的转录水平,表明CsPKCα可能在免疫过程中发挥作用。随后检测了CsPKCα在哈维氏弧菌(Vibrio harveyi)感染后6种免疫相关组织(鳃、肝脏、皮肤、肠、脾脏和肾)中不同时间点的表达水平,发现细菌侵染后24 h或48 h,CsPKCα在鳃、肾脏、皮肤和脾脏中显著上调,侵染后12 h在肝中有显著性下调。研究表明,CsPKCα可能在应对哈维氏弧菌的免疫应答中发挥作用,但是否参与病原菌侵染的巨噬细胞激活以及与T、B细胞相关的信号通路还需要进一步研究。这是关于PKCα参与鱼类细菌性免疫反应的首次报道。
英文摘要:
      Protein kinase C (PKC) plays an important role in T and B lymphocyte-mediated immune response. However, the function of PKC is rarely reported in teleost fish. In this study, we have cloned the PKCα gene (namely CsPKCα), a typical member of the PKC family, and characterized its molecular features and expression pattern. CsPKCα cDNA is 2315 bp in length, with an open reading frame (ORF) of 2013 bp. qRT-PCR analysis showed that CsPKCα was expressed in various tissues, with a high transcription levels in the gill, liver, skin, and intestine, suggesting that it may play a role in immunity. Subsequently, we examined the expression of CsPKCα at different time points in six immune-related tissues (the gill, liver, skin, intestine, spleen, and kidney) after infection with Vibrio harveyi. CsPKCα was significantly upregulated in the gill, kidney, skin, and spleen at 24 h or 48 h post-infection (hpi) and downregulated in the liver at 12 hpi. The results suggested that PKCα might be involved in the immune response to V. harveyi; however, whether it is involved in macrophage activation during bacterial pathogen infection and the signaling pathways associated with T and B cells requires further investigation. This is the first report, to our knowledge, on the involvement of PKCα in the immune response to bacterial infection in fish.
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