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通过分子克隆和表达谱分析,了解两栖气呼吸鲶鱼(Clarias magur,汉密尔顿,1822年)中非哺乳动物Toll样受体22(TLR22)对细菌感染或配体刺激的分子反应。

Understanding the molecular response of non-mammalian toll-like receptor 22 (TLR22) in amphibious air-breathing catfish, Clarias magur (Hamilton, 1822) to bacterial infection or ligand stimulation through molecular cloning and expression profiling.

作者信息

Paria Anutosh, Muduli Chinmayee, Rathore Gaurav

机构信息

ICAR-National Bureau of Fish Genetic Resources, Canal Ring Road, P.O. Dilkusha, Lucknow-226002, Uttar Pradesh, India.

ICAR-National Bureau of Fish Genetic Resources, Canal Ring Road, P.O. Dilkusha, Lucknow-226002, Uttar Pradesh, India.

出版信息

Gene. 2023 May 25;866:147351. doi: 10.1016/j.gene.2023.147351. Epub 2023 Mar 8.

DOI:10.1016/j.gene.2023.147351
PMID:36893873
Abstract

Toll-like receptor (TLR) 22 is a non-mammalian TLR, which is identified initially as a functional substitute of mammalian TLR3 in recognizing cell surface long dsRNA in teleosts. To understand the pathogen surveillance role played by TLR22 in an air-breathing catfish model the full-length cDNA of TLR22 was identified in Clarias magur and found to be consisted of 3597 nucleotides encoding for 966 amino acids. In the deduced amino acid sequence of C. magur TLR22 (CmTLR22) key signature domains such as one signal peptide, 13 LRRs, one transmembrane domain, one LRR_CT domain and an intracellular TIR domain could be identified. The CmTLR22 formed a separate cluster with other catfish TLR22 genes and situated within the TLR22 cluster in the phylogenetic analysis of teleost TLR groups. The CmTLR22 was constitutively expressed in all the 12 tested tissues of healthy C. magur juveniles with the highest transcript abundance in spleen followed by brain, intestine and head kidney. Following induction with the dsRNA viral analogue, poly (I:C), the level of expression of CmTLR22 was up-regulated in tissues such as kidney, spleen and gills. Whereas, in Aeromonas hydrophila-challenged C. magur, the expression levels of CmTLR22 was found to be up-regulated in gills, kidney and spleen, and down-regulated in liver. The findings of the current study suggest that the specific function of TLR22 is evolutionarily conserved in C. magur and might play a key role in mounting immune response by recognizing Gram-negative fish pathogen such as A. hydrophila and aquatic viruses in air-breathing amphibious catfishes.

摘要

Toll样受体(TLR)22是一种非哺乳动物的TLR,最初被鉴定为硬骨鱼中识别细胞表面长双链RNA的哺乳动物TLR3的功能替代物。为了了解TLR22在气呼吸鲶鱼模型中所起的病原体监测作用,在胡鲶中鉴定出了TLR22的全长cDNA,发现其由3597个核苷酸组成,编码966个氨基酸。在胡鲶TLR22(CmTLR22)的推导氨基酸序列中,可以鉴定出关键的特征结构域,如一个信号肽、13个富含亮氨酸重复序列(LRR)、一个跨膜结构域、一个LRR_CT结构域和一个细胞内TIR结构域。在硬骨鱼TLR组的系统发育分析中,CmTLR22与其他鲶鱼TLR22基因形成一个单独的簇,并位于TLR22簇内。CmTLR22在健康胡鲶幼鱼的所有12个测试组织中组成性表达,在脾脏中的转录本丰度最高,其次是脑、肠和头肾。在用双链RNA病毒类似物聚肌胞苷酸(poly (I:C))诱导后,CmTLR22在肾脏、脾脏和鳃等组织中的表达水平上调。然而,在嗜水气单胞菌感染的胡鲶中,发现CmTLR22在鳃、肾脏和脾脏中的表达水平上调,而在肝脏中下调。本研究结果表明,TLR22的特定功能在胡鲶中具有进化保守性,可能通过识别革兰氏阴性鱼类病原体如嗜水气单胞菌和水生病毒,在气呼吸两栖鲶鱼的免疫反应中发挥关键作用。

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