草鱼芳香烃受体ahr1a基因克隆、表达及进化分析
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S917.4

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国家自然科学基金(32030112)


Identification, phylogeny and expression analysis of an aromatic hydrocarbon receptor homologue in grass carp (Ctenopharyngodon idella)
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    摘要:

    芳香烃受体(Aryl hydrocarbon receptor,AhR)作为配体激活的转录因子在T细胞、单核细胞和树突状细胞(Dendritic cells,DCs)的功能调节方面扮演重要角色,炎症相关细胞因子的产生与AhR信号通路也密切相关。为阐明草鱼(Ctenopharyngodon idella)ahr1a基因在炎症发生过程中的作用,本研究克隆获得了草鱼ahr1a基因cDNA序列,全长2 893 bp,其编码区共2 544 bp,编码847个氨基酸。结构域预测发现草鱼AhR1a蛋白含有螺旋-环-螺旋(Helix-loop-helix,HLH)超家族中bHLH-PAS(Period-aryl hydrocarbon receptor nuclear translocator single minded,Pre-Arnt-Sim)亚家族的典型结构域。氨基酸序列分析表明,草鱼AhR1a与黄河鲤(Cyprinus carpio)的相似性与一致性最高。系统进化树结果发现草鱼ahr1a基因与金鱼(Carassius auratus)、黄河鲤和斑马鱼(Danio rerio)的同源基因聚在一支。草鱼ahr1a基因在多个组织(肠、鳃、肾、肝、头肾、胸腺、脾、皮肤、脑)中均有表达,其中肠和鳃表达量最高。草鱼感染柱状黄杆菌(Flavobacterium columnare)后,ahr1a在脾(24、48 h)和头肾(12、24 h)中的表达水平显著下降。腹腔注射草鱼呼肠孤病毒(Reovirus of grass carp,GCRV)后,脾和头肾中ahr1a表达水平无明显变化。体外实验结果显示,脂多糖(Lipopolysaccride,LPS)和IL-1β重组蛋白下调草鱼原代头肾白细胞中ahr1a的表达。综上所述,本研究克隆分析了草鱼ahr1a基因并初步证实其参与炎症反应的调控。

    Abstract:

    The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor and plays an important regulatory role in immune response. The AhR is central to the functions of T cells, monocytes and dendritic cells (DCs) and the production of inflammatory cytokines. To understand the roles of grass carp (Ctenopharyngodon idella) ahr1a gene, we cloned the cDNA of the ahr1a gene in grass carp. The ahr1a cDNA of C. idella is 2 893 bp in length, encoding a protein of 847 aa. It was predicted that grass carp AhR1a protein contains a typical domain of the bHLH-PAS (Period-Aryl hydrocarbon receptor nuclear translocator Single Minded, Pre-Arnt-Sim) subfamily which belongs to the helix-loop-helix (HLX) superfamily. Sequence analysis showed that the amino acid similarity and identity between grass carp AhR1a and common carp (Cyprinus carpio) were the highest. Phylogenetic tree analysis showed that the AhR1a of C. idella was clustered in a clade with homologous amino acid of goldfish (Carassius auratus), common carp and zebrafish (Danio rerio), all of which belong to the cyprinid family. Quantitative real-time PCR (qRT-PCR) analysis found that ahr1a was constitutively expressed in multiple tissues including intestine, gills, kidney, liver, head kidney, thymus, spleen, skin and brain, with the highest expression detected in the intestine and gills. Infection with Flavobacterium columnare resulted in significant decreases of the ahr1a expression in the spleen (24,48 h) and head kidney (12,24 h). In contrast, infection of grass carp reovirus (Reovirus of grass carp, GCRV) did not alter the expression levels of ahr1a in the spleen and head kidney. In the primary head kidney leukocytes, lipopolysaccride (LPS) and recombinant IL-1β dowgregulated the expression of ahr1a. In conclusion, the grass carp ahr1a gene was analyzed in this study. The results suggest that the ahr1a of C. idella may be involved in regulating inflammatory response.

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邬恺正,王俊亚,范丁月,李仕然,徐兢,王梓璇,邹钧.草鱼芳香烃受体ahr1a基因克隆、表达及进化分析[J].上海海洋大学学报,2023,32(1):40-52.
WU Kaizheng, WANG Junya, FAN Dingyue, LI Shiran, XU Jing, WANG Zixuan, ZOU Jun. Identification, phylogeny and expression analysis of an aromatic hydrocarbon receptor homologue in grass carp (Ctenopharyngodon idella)[J]. Journal of Shanghai Ocean University,2023,32(1):40-52.

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  • 收稿日期:2021-11-22
  • 最后修改日期:2022-02-27
  • 录用日期:2022-03-10
  • 在线发布日期: 2023-01-12
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