Tianjin Key Lab of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin, China.
Front Immunol. 2023 Feb 3;14:1098455. doi: 10.3389/fimmu.2023.1098455. eCollection 2023.
The gill of fish is an important immune organ for pathogen defense, but its microRNA (miRNA) expression and regulatory mechanism remain unclear. In this study, we report on the histopathological and immunohistochemical features of the gills of the crucian carp challenged with . Small RNA libraries of the gills were constructed and sequenced on the Illumina HiSeq 2000 platform. A total of 1,165 differentially expressed miRNAs (DEMs) were identified in gills, of which 539 known and 7 unknown DEMs were significantly screened ( < 0.05). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the potential target genes/proteins were primarily involved in 33 immune-related pathways, in which the inflammatory responses were focused on the Toll-like receptor (TLR), mitogen-activated protein kinase (MAPK), and nuclear factor kappa B (NF-κB) signaling pathways. Moreover, the expression levels of 14 key miRNAs (e.g., miR-10, miR-17, miR-26a, miR-144, miR-145, and miR-146a) and their target genes (e.g., , , , , , and ) were verified. In addition, the protein levels based on isobaric tags for relative and absolute quantification (iTRAQ) were significantly associated with the results of the quantitative real-time PCR (qRT-PCR) analysis ( < 0.01). miR-17/pre-miR-17 were identified in the regulation expression of the NF-κB target gene, and the phylogenetic tree analysis showed that the pre-miR-17 of with the closest similarity to the zebrafish is highly conserved in teleosts. This is the first report of the multi-omics analysis of the miRNAs and proteins in the gills of infected with , thus enriching knowledge on the regulation mechanism of the local immune response in Cyprinidae fish.
鱼类的鳃是抵御病原体的重要免疫器官，但它的 microRNA（miRNA）表达和调控机制尚不清楚。在这项研究中，我们报告了鲫鱼鳃在感染后组织病理学和免疫组织化学特征。我们构建了鲫鱼鳃的小 RNA 文库，并在 Illumina HiSeq 2000 平台上进行测序。在鳃中总共鉴定出 1165 个差异表达的 miRNA（DEM），其中 539 个已知和 7 个未知的 DEM 被显著筛选（<0.05）。基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析表明，潜在的靶基因/蛋白主要参与 33 个免疫相关途径，其中炎症反应集中在 Toll 样受体（TLR）、丝裂原激活蛋白激酶（MAPK）和核因子 kappa B（NF-κB）信号通路。此外，还验证了 14 个关键 miRNA（如 miR-10、miR-17、miR-26a、miR-144、miR-145 和 miR-146a）及其靶基因（如 、 、 、 、 ）的表达水平。此外，基于相对和绝对定量同位素标记（iTRAQ）的蛋白质水平与定量实时 PCR（qRT-PCR）分析的结果显著相关（<0.01）。miR-17/pre-miR-17 被鉴定为 NF-κB 靶基因的调节表达，系统发育树分析表明，与斑马鱼 具有最相似的 pre-miR-17 在硬骨鱼中高度保守。这是首次报道感染后鲫鳃中 miRNAs 和蛋白质的多组学分析，丰富了鲤科鱼类局部免疫反应的调控机制知识。
