Jin Ping, Li Shengjie, Sun Lianjie, Lv Caiyun, Ma Fei
Laboratory for Comparative Genomics and Bioinformatics, Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046, China.
Laboratory for Comparative Genomics and Bioinformatics, Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046, China.
Dev Comp Immunol. 2017 Sep;74:243-252. doi: 10.1016/j.dci.2017.05.002. Epub 2017 May 6.
MicroRNAs (miRNAs) are endogenous small non-coding RNAs that participate in diverse biological processes via regulating expressions of target genes at post-transcriptional level. Amphioxus, as modern survivor of an ancient chordate lineage, is a model organism for comparative genomics study. However, miRNAs involved in regulating immune responses in Branchiostoma belcheri are largely unclear. Here, we systematically investigated the microRNAs (miRNAs) involved in regulating immune responses in the cephalochordate amphioxus (Branchiostoma belcheri) through next-generation deep sequencing of amphioxus samples infected with Vibrio parahemolyticus. We identified 198 novel amphioxus miRNAs, consisting of 12 conserved miRNAs, 33 candidate star miRNAs and 153 potential amphioxus-specific-miRNAs. Using microarray profiling, 14 miRNAs were differentially expressed post infection, suggesting they are immune-related miRNAs. Eight miRNAs (bbe-miR-92a-3p, bbe-miR-92c-3p, bbe-miR-210-5p, bbe-miR-22-3p, bbe-miR-1∼bbe-miR-133 and bbe-miR-217∼bbe-miR-216 clusters) were significantly increased at 12 h post-infection, while bbe-miR-2072-5p was downregulated at 6 h and 12 h. Three miRNAs, bbe-miR-1-3p, bbe-miR-22-3p and bbe-miR-92a-3p, were confirmed to be involved in immune responses to infection by qRT-PCR. Our findings further clarify important regulatory roles of miRNAs in the innate immune response to bacterial infection in amphioxus.
微小RNA（miRNA）是内源性小非编码RNA，通过在转录后水平调节靶基因的表达参与多种生物学过程。文昌鱼作为古老脊索动物谱系的现代幸存者，是比较基因组学研究的模式生物。然而，参与调节文昌鱼免疫反应的miRNA在很大程度上尚不清楚。在此，我们通过对感染副溶血性弧菌的文昌鱼样本进行下一代深度测序，系统地研究了头索动物文昌鱼（白氏文昌鱼）中参与调节免疫反应的微小RNA（miRNA）。我们鉴定出198个新的文昌鱼miRNA，包括12个保守miRNA、33个候选星状miRNA和153个潜在的文昌鱼特异性miRNA。使用微阵列分析，14个miRNA在感染后差异表达，表明它们是免疫相关的miRNA。8个miRNA（bbe-miR-92a-3p、bbe-miR-92c-3p、bbe-miR-210-5p、bbe-miR-22-3p、bbe-miR-1至bbe-miR-133以及bbe-miR-217至bbe-miR-216簇）在感染后12小时显著增加，而bbe-miR-2072-5p在6小时和12小时下调。通过qRT-PCR证实3个miRNA，bbe-miR-1-3p、bbe-miR-22-3p和bbe-miR-92a-3p参与了对感染的免疫反应。我们的研究结果进一步阐明了miRNA在文昌鱼对细菌感染的先天免疫反应中的重要调节作用。
