GCRV感染存活草鱼的血清蛋白表达特性

EXPRESSION CHARACTERISTICS OF SERUM PROTEIN IN GRASS CARPS SURVIVING GCRV INFECTION

  • 摘要: 为挖掘草鱼关键抗性分子, 研究利用4D label-free定量蛋白质组学技术系统分析了团队前期获得的GCRV感染存活草鱼(候选抗性草鱼)与对照草鱼的血清蛋白表达差异。共鉴定到858个草鱼血清蛋白, 329个蛋白在两类草鱼中差异表达, 其中163个蛋白在候选抗性草鱼血清中显著高表达, 166个蛋白显著低表达。差异表达蛋白的功能主要注释为体液免疫反应调节相关蛋白, 显著富集到补体凝血级联、细胞黏附和铁死亡等信号通路。对差异表达的免疫相关蛋白进行分析发现, 候选抗性草鱼血清中体液免疫分子MASP2、C4a、C4b、C7b、C8b、C8g、C9、CFI、C3a.1、C3a.2、C3a.3和C3a.6等补体和抗原提呈相关免疫分子MHC1UBA和IGL4V8等蛋白表达水平显著高于对照草鱼, 而T细胞免疫相关分子TRAV、IGHV1-1、IGHV2-1、IGHV6-1、IGHV3-2和IGHV11-2等蛋白表达水平显著低于对照草鱼。免疫印迹检测进一步证实, 以C3和IgM为代表的体液免疫分子在候选抗性草鱼血清中显著高表达, 可能与草鱼抗病能力关联。研究将为高抗性草鱼的选育提供可参考的分子资源。

     

    Abstract: In order to explore the key resistance molecules of grass carp, we compare the serum protein expression profiles of the candidate resistant grass carps and the control grass carps using 4D label-free quantitative proteomics. A total of 858 proteins were identified in the grass carp serum, and 329 proteins were differentially expressed in the candidate resistant grass carps and the control grass carps, with 163 up-regulated and 166 down-regulated in the former. The differentially expressed proteins were mainly related to humoral immune response regulation, including complement coagulation cascade, cell adhesion, iron death and other immune-related signaling pathways. Analysis of differentially expressed immune-related proteins showed that the expression levels of humoral immune molecules such as MASP2, C4a, C4b, C7b, C8b, C8g, C9, CFI, C3a.1, C3a.2, C3a.3 and C3a.6 and antigen presentation related immune molecules including MHC1UBA and IGL4V8, were significantly higher in the serum of candidate resistant grass carps than that in the control grass carps, however, the T cell immunity-related molecules, such as TRAV, IGHV1-1, IGHV2-1, IGHV6-1, IGHV3-2 and IGHV11-2, showed lower expression levels in the serum of candidate resistant grass carps. Notably, Western Blot analysis confirmed that humoral immune molecules, represented by C3 and IgM, were highly expressed in the serum of candidate resistant grass carps, which may be associated with the disease resistance of grass carp. The results may provide theoretical basis and molecular reference for the study of disease-related molecules of grass carp and provide molecular resources for the selective breeding of disease-resistant grass carp.

     

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