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对感染 WSSV 的锯缘青蟹进行高通量测序得到的 miRNA 图谱分析。

The miRNAs profiling revealed by high-throughput sequencing upon WSSV infection in mud crab Scylla paramamosain.

机构信息

Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Institute of Marine Sciences, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.

Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Institute of Marine Sciences, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.

出版信息

Fish Shellfish Immunol. 2020 May;100:427-435. doi: 10.1016/j.fsi.2020.03.002. Epub 2020 Mar 5.

Abstract

microRNAs (miRNAs) are known to regulate various immune functions by silencing the target genes in both vertebrates and invertebrates. However, in mud crab Scylla paramamosain, the role of miRNAs during the response to virus invasion remains unclear. To investigate the roles of miRNAs in S. paramamosain during virus infection, the mud crab was challenged with white spot syndrome virus (WSSV) and then subjected to the transcriptional analysis at different conditions. The results of high-throughput sequencing revealed that 940,379 and 1,306,023 high-quality mappable reads were detected in the hemocyte of normal and WSSV-infected mud crabs, respectively. Besides, the total number of 261 unique miRNAs were identified. Among them, 131 miRNAs were specifically expressed in the hemocytes of normal mud crabs, 46 miRNAs were specifically transcribed in those of WSSV-infected individuals, the other 84 miRNAs were expressed in both normal and WSSV-infected individuals. Furthermore, a number of 152 (89 down-regulated and 63 up-regulated) miRNAs were found to be differentially expressed in the WSSV-infected hemocytes, normalized to the controls. The identified miRNAs were subjected to GO analysis and target gene prediction and the results suggested that the differentially regulated miRNAs were mainly correlated with the changes of the immune responses of the hemocytes, including phagocytosis, melanism, and apoptosis as well. Taken together, the results demonstrated that the expressed miRNAs during the virus infection were mainly involved in the regulation of immunological pathways in mud crabs. Our findings not only enrich the understanding of the functions of miRNAs in the innate immune system but also provide some novel potential targets for the prevention of WSSV infection in crustaceans.

摘要

微小 RNA(miRNAs)被认为通过在脊椎动物和无脊椎动物中沉默靶基因来调节各种免疫功能。然而,在泥蟹 Scylla paramamosain 中,miRNAs 在病毒入侵时的作用尚不清楚。为了研究 miRNAs 在泥蟹 S. paramamosain 感染病毒过程中的作用,用白斑综合征病毒(WSSV)对泥蟹进行了攻毒,然后在不同条件下进行转录分析。高通量测序的结果显示,正常和 WSSV 感染的泥蟹血细胞中分别检测到 940,379 和 1,306,023 个高质量可映射读数。此外,鉴定出 261 个独特的 miRNAs。其中,131 个 miRNAs 特异性表达在正常泥蟹的血细胞中,46 个 miRNAs 特异性转录在 WSSV 感染个体的血细胞中,其余 84 个 miRNAs 在正常和 WSSV 感染个体中均表达。此外,在 WSSV 感染的血细胞中发现 152 个(89 个下调和 63 个上调)miRNAs 的表达水平与对照组相比发生了差异。对鉴定出的 miRNAs 进行 GO 分析和靶基因预测,结果表明差异表达的 miRNAs 主要与血细胞免疫反应的变化相关,包括吞噬作用、黑化作用和凋亡等。总之,研究结果表明,病毒感染期间表达的 miRNAs 主要参与泥蟹免疫途径的调节。本研究不仅丰富了 miRNAs 在先天免疫系统中的功能理解,还为甲壳动物 WSSV 感染的预防提供了一些新的潜在靶标。

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