Deep Illumina sequencing reveals conserved and novel microRNAs in grass carp in response to grass carp reovirus infection.
Deep Illumina sequencing reveals conserved and novel microRNAs in grass carp in response to grass carp reovirus infection.
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DOI：
10.1186/s12864-017-3562-4
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发表时间：
2017-02-20
期刊：
影响因子：
4.4
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
He L;Zhang A;Chu P;Li Y;Huang R;Liao L;Zhu Z;Wang Y
由草鱼呼肠孤病毒（GCRV）引起的草鱼出血病是影响草鱼养殖业发展的主要疾病。GCRV的发病机制和出血症状仍不清楚。microRNAs（miRNAs）是参与多种生物学过程的关键调控因子。本研究的目的是鉴定草鱼对GCRV感染应答的保守和新的miRNAs，并试图揭示GCRV致病和出血症状的机制。用GCRV感染草鱼，并在感染后0（对照）、1、3、5、7和9天（dpi）采集脾脏样品。这些样品用于构建和测序小RNA文库。共鉴定了1208个miRNAs，其中278个为已知miRNAs，930个为新发现的miRNAs。与对照组相比，36个miRNAs被鉴定为表现出差异表达，并且预测了36个miRNAs的536个靶基因。这些靶基因的GO和KEGG富集分析显示，许多显著富集的术语与免疫应答、血液凝固、止血以及补体和凝血级联相关，尤其是GO术语“血液凝固”和途径“补体和凝血级联”。选择了10个参与“补体和凝血级联”的代表性靶基因进行qPCR分析，结果显示这些靶基因的表达模式在7 dpi时显著上调，表明“补体和凝血级联”途径被强烈激活。在草鱼中发现了与GCRV感染相关的保守和新的miRNAs，其中已知miRNAs 278个，新发现miRNAs 930个。许多靶基因参与免疫应答、凝血、止血以及补体和凝血级联反应。途径“补体和凝血级联反应”的强烈激活可能导致内皮细胞和血细胞损伤和出血症状。本研究为理解GCRV的发病机制和出血症状提供了新的见解。本文的在线版本（doi：10.1186/s12864-017-3562-4）包含补充材料，可供授权用户使用。
The grass carp hemorrhagic disease caused by the grass carp reovirus (GCRV) is a major disease that hampers the development of grass carp aquaculture. The mechanism underlying GCRV pathogenesis and hemorrhagic symptoms is still unknown. MicroRNAs (miRNAs) are key regulators involved in various biological processes. The aim of this study was to identify conserved and novel miRNAs in grass carp in response to GCRV infection, as well as attempt to reveal the mechanism underlying GCRV pathogenesis and hemorrhagic symptoms. Grass carp were infected with GCRV, and spleen samples were collected at 0 (control), 1, 3, 5, 7, and 9 days post-infection (dpi). These samples were used to construct and sequence small RNA libraries. A total of 1208 miRNAs were identified, of which 278 were known miRNAs and 930 were novel miRNAs. Thirty-six miRNAs were identified to exhibit differential expression when compared with the control, and 536 target genes were predicted for the 36 miRNAs. GO and KEGG enrichment analyses of these target genes showed that many of the significantly enriched terms were associated with immune response, blood coagulation, hemostasis, and complement and coagulation cascades, especially the GO term “blood coagulation” and pathway “complement and coagulation cascades.” Ten representative target genes involved in “complement and coagulation cascades” were selected for qPCR analysis, and the results showed that the expression patterns of these target genes were significantly upregulated at 7 dpi, suggesting that the pathway “complement and coagulation cascades” was strongly activated. Conserved and novel miRNAs in response to GCRV infection were identified in grass carp, of which 278 were known miRNAs and 930 were novel miRNAs. Many of the target genes involved in immune response, blood coagulation, hemostasis, and complement and coagulation cascades. Strong activation of the pathway “complement and coagulation cascades” may have led to endothelial-cell and blood-cell damage and hemorrhagic symptoms. The present study provides a new insight into understanding the mechanism underlying GCRV pathogenesis and hemorrhagic symptoms. The online version of this article (doi:10.1186/s12864-017-3562-4) contains supplementary material, which is available to authorized users.