路俊怡, 姜立妍, 龙凯, 王韬, 吴正理, 李艳红. HcTLR1通过MyD88-NF-κB信号通路参与三角帆蚌抗菌免疫应答[J]. 南方水产科学. DOI: 10.12131/20240093
引用本文: 路俊怡, 姜立妍, 龙凯, 王韬, 吴正理, 李艳红. HcTLR1通过MyD88-NF-κB信号通路参与三角帆蚌抗菌免疫应答[J]. 南方水产科学. DOI: 10.12131/20240093
LU Junyi, JIANG Liyan, LONG Kai, WANG Tao, WU Zhengli, LI Yanhong. HcTLR1 involved in antimicrobial immune response by MyD88-NF-κB signaling pathway in Hyriopsis cumingii[J]. South China Fisheries Science. DOI: 10.12131/20240093
Citation: LU Junyi, JIANG Liyan, LONG Kai, WANG Tao, WU Zhengli, LI Yanhong. HcTLR1 involved in antimicrobial immune response by MyD88-NF-κB signaling pathway in Hyriopsis cumingii[J]. South China Fisheries Science. DOI: 10.12131/20240093

HcTLR1通过MyD88-NF-κB信号通路参与三角帆蚌抗菌免疫应答

HcTLR1 involved in antimicrobial immune response by MyD88-NF-κB signaling pathway in Hyriopsis cumingii

  • 摘要: Toll样受体 (Toll-like receptors, TLR) 家族是一类进化保守的病原体识别受体,在检测和防御微生物病原体的先天免疫中发挥着重要作用。为了探究HcTLR1基因在三角帆蚌 (Hyriopsis cumingii) 抗菌应答中的作用,采用cDNA末端快速克隆技术 (Rapid-amplification of cDNA ends, RACE) 获得HcTLR1基因cDNA全长序列,采用实时荧光定量PCR分析比较HcTLR1基因在三角帆蚌不同组织和不同免疫刺激后的表达水平,利用双链RNA干扰技术分析敲低该基因后MyD88依赖性通路及相关免疫基因的变化。结果显示,HcTLR1基因开放阅读框全长为3 687 bp,编码1 228个氨基酸。预测的HcTLR1蛋白结构包含多个亮氨酸富集的重复序列 (Leucine-rich repeat, LRR) 结构域、1个跨膜结构域和1个胞内Toll/白细胞介素-1 (Toll-IL-1 receptor domain, TIR) 受体。此外,HcTLR1基因在血细胞中表达量最高,且对维氏气单胞菌GL (Aeromonas veronii GL1) 和病原体相关分子模式 (Pathogen-associated molecular patterns, PAMPs) 刺激呈现出差异显著的时间依赖性变化。敲低HcTLR1基因后,维氏气单胞菌GL1刺激所激活的MyD88相关通路基因、抗菌肽、溶菌酶、防御素、乳清酸性蛋白、脂多糖结合蛋白/杀菌通透性增加蛋白2和白细胞介素17等基因的表达水平显著降低。结果表明,HcTLR1参与三角帆蚌对于微生物感染过程中MyD88依赖性信号通路活化并促进血细胞抵抗机制。

     

    Abstract: Toll-like receptor (TLR) family is an evolutionarily conserved pathogen recognition receptor, playing an important role in detecting and defending against microbial pathogens. To investigate the role of the HcTLR1 gene in the antimicrobial response of Hyriopsis cumingii, the full-length cDNA sequence of the HcTLR1 gene was cloned using rapid-amplification of cDNA ends (RACE) technology; real-time fluorescence quantitative PCR analysis was employed to compare the expression levels of the HcTLR1 gene in various tissues of H. cumingii challenged with different stimuli; double-stranded RNA interference technology was used to analyze the changes in the MyD88-dependent pathway and related immune genes after the knockdown of the gene. The results show that the open reading frame (ORF) of the HcTLR1 gene was 3 687 bp, encoding 1 228 amino acids. The predicted structure of the HcTLR1 protein contained multiple Leucine-rich repeat domains, a transmembrane domain and an intracellular Toll/interleukin-1 receptor. Furthermore, the mRNA expression of HcTLR1 gene was highest in hemocytes and exhibited significant changes in response to Aeromonas veronii GL1 and pathogen-associated molecular patterns stimulation at different time points. Moreover, the knockdown of the HcTLR1 gene significantly reduced the expression levels of genes in MyD88-related pathway, antibacterial peptides, lysozyme, defensins, lactoferrin, LPS1-binding protein/bactericidal permeability-increasing protein 2, and interleukin 17 stimulated by A. veronii GL1. In conclusion, it is suggested that HcTLR1 activates MyD88-dependent signaling pathways in H. cumingii during microbial infection and promotes resistance mechanisms in hemocytes.

     

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