草鱼呼肠孤病毒Ⅱ型(GCRVⅡ)可引起草鱼严重出血性疾病,影响我国养殖业。已从中国不同流行地区收集了GCRVⅡ分离株,这些分离物可以导致草鱼不同程度的出血症状。稀有小鱼(Gobiociprisrarus)因其对GCRV的高度敏感性而被广泛用作模型鱼研究出血性疾病的机制。在这项研究中,使用基于相对和绝对定量(iTRAQ)的定量蛋白质组学的同量异序标签,研究了罕见小鱼感染GCRV毒力分离株JZ809和减毒分离株XT422后脾脏中的蛋白质水平.获得了毒力和减毒感染组的109和50种差异表达蛋白(DEPs),分别,其中两组共确定了40个DEP.将蛋白质表达谱与基因本体论(GO)注释相结合,稀有小鱼对两种基因型GCRVⅡ在上调蛋白方面的反应相似,专注于ATP合成,其中ATP可以作为“危险”信号激活真核生物中的免疫反应。同时,毒力基因型JZ809可诱导更多的免疫蛋白并增加泛素-蛋白酶体系统成员的水平以适应病毒感染。然而,伴随着持续和过度的炎症反应和不断下降的碳代谢,与减毒XT422相比,JZ809感染后,罕见小鱼呈现更严重的出血性疾病和死亡率.该结果提供了有价值的信息,将增加我们对具有不同毒力水平的病毒的发病机理以及病毒与宿主之间相互作用的机制的理解。此外,在XT422感染组中仅显著上调的6种蛋白均属于第2簇,在JZ809感染组中仅上调的30种蛋白中,有28种被聚集到第1簇.对于下调的蛋白质,XT422感染组的所有DEP均聚集到第4簇,在JZ809感染组中仅显著下调的39种蛋白质中,有25种属于第3簇.结果表明,减毒XT422感染组中的DEP可能是敏感的,当鱼类经历病毒感染时,它们的丰度变化更快。
Grass carp reovirus genotype Ⅱ (GCRV II) causes severe hemorrhagic disease in grass carp and affects the aquaculture industry in China. GCRV Ⅱ isolates have been collected from different epidemic areas in China, and these isolates can lead to different degrees of hemorrhagic symptoms in grass carp. Rare minnow (Gobiocypris rarus) is widely used as a model fish to study the mechanism of hemorrhagic disease because of its high sensitivity to GCRV. In this study, the protein levels in the spleen of rare minnow after infection with GCRV virulent isolate JZ809 and attenuated isolate XT422 were investigated using isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics. 109 and 50 differentially expressed proteins (DEPs) in the virulent and attenuated infection groups were obtained, respectively, among which 40 DEPs were identified in both groups. Combining protein expression profiling with gene ontology (GO) annotation, the responses of rare minnow to the two genotypes GCRV Ⅱ in terms of upregulated proteins were similar, focusing on ATP synthesis, in which ATP can serve as a \"danger\" signal to activate an immunoreaction in eukaryotes. Meanwhile, the virulent genotype JZ809 induced more immunoproteins and increased the levels of ubiquitin-proteasome system members to adapt to virus infection. However, together with a persistent and excessive inflammatory response and declining carbon metabolism, rare minnow presented more severe hemorrhagic disease and mortality after infection with virulent JZ809 than with attenuated XT422. The results provide a valuable information that will increase our understanding of the pathogenesis of viruses with different levels of virulence and the mechanism of interaction between the virus and host. Furthermore, the 6 proteins that were only significantly upregulated in the XT422 infection group all belonged to cluster 2, and 28 of 30 proteins that were only upregulated in JZ809 infection group were clustered into cluster 1. For the downregulated proteins, all DEPs in the XT422 infection group were clustered into cluster 4, and 25 of 39 proteins that were only significantly downregulated in the JZ809 infection group belonged to cluster 3. The results indicated that the DEPs in the attenuated XT422 infection group might be sensitive and their abundance changed more quickly when fish experienced virus infection.