Du Xiaogang, Li Yunkun, Li Dong, Lian Fangliang, Yang Shiyong, Wu Jiayun, Liu Hanmei, Bu Guixian, Meng Fengyan, Cao Xiaohan, Zeng Xianyin, Zhang Huaiyu, Chen Zhiyu
Department of Engineering and Applied Biology, College of Life Science, Sichuan Agricultural University, Ya'an 625014, Sichuan, China.
Department of Engineering and Applied Biology, College of Life Science, Sichuan Agricultural University, Ya'an 625014, Sichuan, China.
Fish Shellfish Immunol. 2017 Mar;62:13-23. doi: 10.1016/j.fsi.2017.01.004. Epub 2017 Jan 4.
Schizothorax prenanti (S. prenanti) is an important economical cold-water fish species in southwestern China, but it is susceptible to various pathogens infection. In order to clearly elucidate the antiviral mechanism, in this study, we have analyzed the transcriptome of S. prenanti spleen after challenge with the virus mimic, poly (I:C) (pIC), using next generation sequencing technology (RNA-seq). A total of 313 differential expressed genes (DEGs) in spleen at 12 h were obtained after pIC treatment, including 268 significantly up-regulated unigenes (fold change > 2) and 45 significantly down-regulated unigenes (fold change > 2). Through the immune-related DEGs (IRDs) screening, 47 IRDs were used to establish heat map, which intuitively showed a significantly difference after pIC treatment. To validate the RNA-seq data and observe gene expression, the expression levels of 14 IRDs were detected by qPCR after pIC treatment at 0, 4, 8, 12, and 24 h. The results indicated that the qPCR data presented a positive line correlation with RNA-seq data, and the 14 IRDs were responsive to pIC stimulation except IL-1β. Thus, based on the RNA-seq and qPCR data, we inferred that MDA5- and Jak-mediated signaling pathways may involve in the antiviral signaling transduction, and induce type I IFNs and ISGs to block virus invasion, respectively. Unfortunately, TLR3 and TLR22, as receptors of virus dsRNA, were no significantly expressed in this study. Nonetheless, our study still provides useful mRNA sequences of antiviral immunity for further immunological research, and facilitates improving disease restriction in S. prenanti.
齐口裂腹鱼是中国西南部一种重要的经济冷水鱼类,但它易受各种病原体感染。为了清楚地阐明其抗病毒机制,在本研究中,我们使用下一代测序技术(RNA测序)分析了用病毒模拟物聚肌苷酸-聚胞苷酸(pIC)刺激后齐口裂腹鱼脾脏的转录组。pIC处理后,在12小时时脾脏中共获得313个差异表达基因(DEG),包括268个显著上调的单基因(倍数变化>2)和45个显著下调的单基因(倍数变化>2)。通过免疫相关DEG(IRD)筛选,47个IRD用于建立热图,直观地显示了pIC处理后的显著差异。为了验证RNA测序数据并观察基因表达,在pIC处理后0、4、8、12和24小时通过qPCR检测了14个IRD的表达水平。结果表明,qPCR数据与RNA测序数据呈正线性相关,除IL-1β外,14个IRD对pIC刺激有反应。因此,基于RNA测序和qPCR数据,我们推断MDA5和Jak介导的信号通路可能参与抗病毒信号转导,并分别诱导I型干扰素和干扰素刺激基因(ISG)来阻断病毒入侵。不幸的是,作为病毒双链RNA受体的TLR3和TLR22在本研究中无显著表达。尽管如此,我们的研究仍为进一步的免疫学研究提供了有用的抗病毒免疫mRNA序列,并有助于改善齐口裂腹鱼的疾病防控。