Transcriptomic analysis of exosomal shuttle mRNA in Pacific oyster Crassostrea gigas during bacterial stimulation
Transcriptomic analysis of exosomal shuttle mRNA in Pacific oyster Crassostrea gigas during bacterial stimulation
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细菌刺激期间太平洋牡蛎外泌体穿梭 mRNA 的转录组分析
DOI：
10.1016/j.fsi.2018.01.017
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发表时间：
2018-03
影响因子：
4.7
通讯作者：
Lei Wang
中科院分区：
文献类型：
--
作者：
Mengqiang Wang;Mei Liu;Baojie Wang;Keyong Jiang;Zhihao Jia;Lingling Wang;Lei Wang
作为海洋无脊椎动物，牡蛎缺乏适应性免疫，而是将先天免疫作为抵御入侵者的第一道防线，几乎也是唯一的防御机制。越来越多的研究成果表明，外泌体可作为先天免疫效应物，有助于宿主防御机制。为了更好地了解长牡蛎外泌体在应对细菌刺激时的免疫功能，本研究应用RNA测序技术，探究金黄色葡萄球菌和灿烂弧菌刺激后牡蛎外泌体的整体表达变化。通过Ion Torrent Proton测序共产生171573691条单端原始读数，经修剪后得到121988325条纯净读数，随后鉴定出1505个丰富的外泌体穿梭信使核糖核酸（esmRNAs）。基因本体论（GO）分析显示，这些丰富的esmRNAs可分为15种细胞成分、12种分子功能和21种生物学过程，并且通过京都基因与基因组百科全书（KEGG）将这些丰富的esmRNAs定位到62条生物信号通路上。在金黄色葡萄球菌刺激组与对照组之间，共鉴定出68个显著差异表达基因（DEGs，倍数变化≥2，Q值<0.05），其中21个上调，47个下调。而在灿烂弧菌刺激组与对照组之间，鉴定出99个显著差异表达基因，其中42个上调，57个下调。为验证转录组数据，随机选取24个差异表达基因，通过实时荧光定量聚合酶链反应（qRT-PCR）进行确认，结果表明它们的表达模式与RNA测序分析结果高度吻合。本研究将丰富长牡蛎转录组数据库，并为深入了解牡蛎外泌体在抵御细菌感染中的免疫功能提供思路。
As marine invertebrates, oysters lack adaptive immunity and employ innate immunity as the front line and almost the solo defense mechanism to protect them against invaders. Accumulating research achievements demonstrated that exosomes could act as innate immune effectors that contribute to host defense mechanism. To better understand the immune functions of exosomes in Crassostrea gigas against bacterial stimulation, RNA-Seq was applied to explore the global expression changes of exosomes in oyster after Staphylococcus aureus and Vibrio splendidus stimulation. Totally 171573691 single end raw reads were yielded via Ion Torrent Proton sequencing, which were trimmed into 121988325 clean reads, and then 1505 abundant exosomal shuttle mRNAs (esmRNAs) were identified. Gene ontology (GO) analysis revealed that these abundant esmRNAs could be categorized into 15 cellular components, 12 molecular functions and 21 biological processes, and these abundant esmRNAs were mapped onto 62 biological signaling pathways by KEGG. In total, 68 significant differentially expressed genes (DEGs, Fold change≥2, Q-value < 0.05) were identified between S. aureus stimulated group and control group, including 21 up-regulated and 47 downregulated ones. While 99 significant DEGs between V. splendidus challenged group and control group were identified, including 42 up-regulated and 57 down-regulated ones. To validate the transcriptomic data, 24 DEGs were randomly selected and confirmed via quantitative real-time PCR (qRT-PCR) and the results showed that their expression patterns agreed well with the RNA-Seq analysis. This study would enrich the C.gigas transcriptome database and provide insight into the immune functions of oyster exosomes against bacterial infection.
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影响因子：
46.9
作者：
Trapnell C;Williams BA;Pertea G;Mortazavi A;Kwan G;van Baren MJ;Salzberg SL;Wold BJ;Pachter L
通讯作者：
Pachter L
影响因子：
13.3
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影响因子：
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作者：
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通讯作者：
Zhang Guofan
影响因子：
82.9
作者：
Zitvogel, L;Regnault, A;Amigorena, S
通讯作者：
Amigorena, S
影响因子：
4.7
作者：
Mengqiang Wang;Lingling Wang;Ying Guo;Zhi Zhou;Qilin Yi;Daoxiang Zhang;Huan Zhang;Rui Liu
通讯作者：
Mengqiang Wang;Lingling Wang;Ying Guo;Zhi Zhou;Qilin Yi;Daoxiang Zhang;Huan Zhang;Rui Liu