Molecular characterization and expression of TLR7 and TLR8 in barbel chub (Squaliobarbus curriculus): Responses to stimulation of grass carp reovirus and lipopolysaccharide
Molecular characterization and expression of TLR7 and TLR8 in barbel chub (Squaliobarbus curriculus): Responses to stimulation of grass carp reovirus and lipopolysaccharide
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鲢鱼 (Squaliobarbus curriculus) 中 TLR7 和 TLR8 的分子特征和表达：对草鱼呼肠孤病毒和脂多糖刺激的反应
DOI：
10.1016/j.fsi.2018.09.035
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发表时间：
2018-09
影响因子：
4.7
通讯作者：
Tiaoyi Xiao
中科院分区：
文献类型：
--
作者：
Shengzhen Jin;Xin Zhao;Hongquan Wan;Jianming Su;Jing'an Wang;Chunhua Ding;Yaoguo Li;Tiaoyi Xiao
黄颡鱼（Squaliobarbus）是一种广泛分布于亚洲的小型经济鱼类，具有较强的抗病能力。本研究对S.克隆了草鱼ScTLR 7和ScTLR 8基因，研究了它们在结构上的差异以及对草鱼病毒（GCRV）感染和脂多糖刺激的反应。ScTLR 7 cDNA全长3715 bp，包含一个3162 bp的开放阅读框，编码1053个氨基酸残基。ScTLR 8全长4624 bp，包含一个3072 bp的开放阅读框，编码1023个氨基酸残基; ScTLR 7和ScTLR 8均由N端信号肽、富含亮氨酸重复序列（LRR）和Toll/IL-1受体结构域组成。ScTLR 7和ScTLR 8的LRR基序弯曲成马蹄铁状螺线管结构，但两个基因的LRR数目不同。系统发育分析表明，ScTLR 7和ScTLR 8与草鱼和团头鲂的亲缘关系较近。实时荧光定量聚合酶链反应（PCR）分析显示，ScTLR 7在S.结果表明，脑中的脂肪酸含量最高，其次是脾和心，肠中的脂肪酸含量最低。ScTLR 8在脾脏中的表达量最高，在肝脏中的表达量最低。LPS刺激后，ScTLR 7和ScTLR 8的相对表达水平总体呈上调趋势。LPS刺激后12、24、72和168 h，I-IFN的表达水平与6 h相比，总体呈下调趋势。与GCRV感染后6 h相比，ScTLR 7在感染后12 ~ 168 h转录水平上调，ScTLR 8、MyD 88和I-IFN在感染后12 ~ 72 h转录水平先上调，72 ~ 168 h下调。相关性分析显示，GCRV刺激后，ScTLR 7与MyD 88的表达量呈显著正相关（Pearson相关系数：0.909，P：0.033），MyD 88与I型IFN的表达量呈显著正相关（Pearson相关系数：0.962，P：0.009）。这些结果表明ScTLR 7和ScTLR 8在草鱼感染和脂多糖刺激的应答中可能起重要作用，并触发不同的下游免疫信号通路。
The barbel chub (Squaliobarbus curriculus) is a kind of small size commercial fish species that is widely spread in Asia and has shown significant resistance to disease. In this study, the full-length cDNA sequences ofToll-like receptor(TLR)7and8fromS. curriculus, designated asScTLR7andScTLR8, were cloned, and their differences in the structure and the responses to the grass carp (GCRV) infection and lipopolysaccharide stimulation were investigated. The full-length 3715 base pair (bp) cDNA ofScTLR7contained a complete open reading frame of 3162 bp and encoded a putative polypeptide of 1053 amino acid residues. The full-length 4624 base pair (bp) cDNA ofScTLR8contained a complete open reading frame of 3072 bp and encoded a putative polypeptide of 1023 amino acid residues.ScTLR7andScTLR8consisted of N-terminal signal peptide, leucine-rich repeats (LRRs), and Toll/IL-1 Receptors domain. LRR motifs ofScTLR7andScTLR8bend into horseshoe-like solenoid structure, while the number of LRRs between the two genes is different. Phylogenetic analysis showed that both theScTLR7andScTLR8were closely clustered withCtenopharyngodon idellusandMegalobrama amblycephala. Quantitative real-time polymerase chain reaction analysis showed that the expression levels ofScTLR7inS. curriculuswere most abundant in the brain followed by the spleen and heart, and the lowest in the intestine. The highest expression level ofScTLR8was observed in spleen and the lowest in the liver. After LPS stimulation, the relative expression levels of bothScTLR7andScTLR8exhibited an overall trend of up-regulation. The expression levels oftype I-IFNshowed an overall trend of down-regulation at time points of 12, 24, 72 and 168 h compared to that of 6 h after LPS stimulation. Compared to 6 h post GCRV infection, the transcription level ofScTLR7was up-regulated from 12 to 168 h, and transcription levels ofScTLR8,MyD88, andtype I-IFNwere firstly up-regulated from 12 to 72 h, and then down-regulated from 72 to 168 h. Correlation analysis showed that expression level ofScTLR7in the spleen was significantly positively correlated with that ofMyD88(Pearson correlation coefficient: 0.909,P: 0.033), and a significantly positive correlation was also observed between expression levels ofMyD88andtype I IFN(Pearson correlation coefficient: 0.962,P: 0.009), after GCRV stimulation. These results indicate thatScTLR7andScTLR8may play important roles in the responses to the grass carp (GCRV) infection and lipopolysaccharide stimulation and trigger different downstream immune signal pathways.
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DOI：
10.1353/ccr.2015.0004
发表时间：
2015-07
期刊：
Cream City Review
影响因子：
--
作者：
Trace Peterson
通讯作者：
Trace Peterson
DOI：
10.1016/b978-012455900-4/50268-3
发表时间：
2005-01-01
期刊：
MEASURING IMMUNITY: BASIC BIOLOGY AND CLINICAL ASSESSMENT
影响因子：
--
作者：
Hawn, Thomas R.;Underhill, David M.
通讯作者：
Underhill, David M.
DOI：
10.1007/978-1-4419-5906-5_77
发表时间：
2010-09
期刊：
Composites Part B: Engineering
影响因子：
--
作者：
Carlisle M. Adams
通讯作者：
Carlisle M. Adams
影响因子：
13.6
作者：
Jiménez-Dalmaroni MJ;Gerswhin ME;Adamopoulos IE
通讯作者：
Adamopoulos IE
影响因子：
4.7
作者：
Youchuan Wei;Shu Hu;Baobao Sun;Qihuan Zhang;G. Qiao;Zisheng Wang;Rong Shao;Guoqiang Huang;Z. Qi
通讯作者：
Youchuan Wei;Shu Hu;Baobao Sun;Qihuan Zhang;G. Qiao;Zisheng Wang;Rong Shao;Guoqiang Huang;Z. Qi