TLRs基因家族在鸭法氏囊中的表达模式及分子进化分析

doi:10.11843/j.issn.0366-6964.2018.09.006

摘要/Abstract

摘要：

旨在探究TLRs基因家族（toll-like receptors，TLRs）在鸭法氏囊中的表达情况和功能差异。本研究利用qRT-PCR检测鸭法氏囊胚后发育中TLRs基因家族的表达情况，对表达模式进行聚类分析，并分别构建了TLRs启动子区和编码区的系统进化树。结果显示，TLR2a在鸭法氏囊2周龄（W2）和6周龄（W6）时期的表达水平显著高于4周龄（W4）时期（P<0.05），其余TLRs各成员在鸭法氏囊胚后发育各阶段的表达差异不显著（P>0.05）；TLR1a、TLR2a、TLR2b以及TLR3、TLR5、TLR15具有相似的表达模式；在编码区进化树上TLR1a、TLR1b、TLR2a、TLR2b聚为一支，与TLR15距离较近，TLR3和TLR5在同一分支上，TLR4和TLR21为单独两个分支，TLR21与其他TLRs成员距离最远；而TLRs基因家族启动子区进化树没有明显规律。结果提示，TLRs家族成员可能不参与鸭法氏囊胚后发育调控过程；TLR2a和TLR2b，以及TLR3、TLR5和TLR15可能具有相似功能，TLR1a和TLR1b虽然具有较高的序列相似性，但可能在进化中出现了功能分化。TLRs表达模式与启动子区序列进化无明显联系。

Abstract:

This study aimed to explore the expression and function of TLRs gene family in duck bursa of Fabricius, therefore, the mRNA expressions of TLRs genes in duck bursa of Fabricius during the postembryonic development stage were detected by qRT-PCR. The expression patterns were analyzed by clustering and the phylogenetic trees based on the TLRs promoters and coding regions were constructed, respectively. The results showed that the expression level of TLR2a in duck bursa of Fabricius was significantly higher in W2 and W6 than in W4(P<0.05). The expressions of other TLRs in the different postembryonic development stages of duck bursa of Fabricius were not significantly different (P>0.05). TLR1a, TLR2a and TLR2b had similar expression patterns, as well as TLR3, TLR5 and TLR15. In the phylogenetic tree of the coding region, TLR1a, TLR1b, TLR2a and TLR2b were clustered together, and they were closer to TLR15. TLR3 and TLR5 were on the same branch, TLR4 and TLR21 were on two separate branches, respectively, and TLR21 was far away from the other TLRs. However, there was no obvious rules in the phylogenetic tree of TLRs gene family promoter regions. The results indicate that TLRs family members may not be involved in the regulation of postembryonic development in duck bursa of Fabricius; TLR2a and TLR2b may have similar function, as well as TLR3, TLR5 and TLR15. Although TLR1a and TLR1b have high sequence similarity, functional differentiation may occur during the long process of evolution. There was no significant relationship between TLRs expression patterns and its promoter region sequence evolution.

中图分类号:

S834.2

甘心梦, 陈凯文, 刘贺贺, 李彦莹, 王继文, 李亮, 白丽丽. TLRs基因家族在鸭法氏囊中的表达模式及分子进化分析[J]. 畜牧兽医学报, 2018, 49(9): 1851-1860.

GAN Xin-meng, CHEN Kai-wen, LIU He-he, LI Yan-ying, WANG Ji-wen, LI Liang, BAI Li-li. The Expression Patterns of TLRs Gene Family during Duck Bursa of Fabricius Development and Their Molecular Evolutionary Analysis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1851-1860.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.09.006

https://www.xmsyxb.com/CN/Y2018/V49/I9/1851

参考文献

[1] ROACH J C,GLUSMAN G,ROWEN L,et al.The evolution of vertebrate Toll-like receptors[J].Proc Natl Acad Sci USA, 2005,102(27):9577-9582.
[2] AKIRA S.Toll receptor families:structure and function[J].Semin Immunol,2004,16(1):1-2.
[3] 郭佳佳,戴子淳,陆晨,等. TLR家族基因在产蛋高峰期和停产期鸭肠道中的表达研究[J]. 畜牧兽医学报, 2016, 47 (10):2012-2019.
GUO J J, DAI Z C, LU C, et al. Expression of toll-like receptors in duck intestinal tract at the peak and end of lay[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47 (10):2012-2019. (in Chinese)
[4] WANG J L,ZHANG Z,LIU J,et al.Ectodomain architecture affects sequence and functional evolution of vertebrate toll-like receptors[J].Sci Rep,2016,6:26705.
[5] TRINCHIERI G,SHER A.Cooperation of Toll-like receptor signals in innate immune defence[J].Nat Rev Immunol,2007,7(3):179-190.
[6] BUCKLEY K M,RAST J P.Dynamic evolution of toll-like receptor multigene families in echinoderms[J].Front Immunol,2012,3:136.
[7] JIA H,LI G,LI J,et al.Cloning,expression and bioinformatics analysis of the duck TLR 4 gene[J].Br Poult Sci,2012,53(2):190-197.
[8] KANNAKI T R,REDDY M R,VERMA P C,et al.Differential toll-like receptor (TLR) mRNA expression patterns during chicken embryological development[J].Anim Biotechnol,2015,26(2):130-135.
[9] SONG C P,YU S Q,DUAN Y B,et al.Effect of age on the pathogenesis of DHV-1 in Pekin ducks and on the innate immune responses of ducks to infection[J].Arch Virol,2014,159(5):905-914.
[10] GONG Y W,FENG S S,LI S Q,et al.Genome-wide characterization of Toll-like receptor gene family in common carp (Cyprinus carpio) and their involvement in host immune response to Aeromonas hydrophila infection[J].Comp Biochem Physiol Part D Genomics Proteomics,2017,24:89-98.
[11] ANSARI A R,WEN L,HUANG H B,et al.Lipopolysaccharide stimulation upregulated toll-like receptor 4 expression in chicken cerebellum[J].Vet Immunol Immunopathol,2015,166(3-4):145-150.
[12] 刘俊莹.两品种鸭法氏囊差异表达基因、miRNA的富集及验证[D].雅安:四川农业大学,2015.
LIU J Y.Enrichment and verification of differentially expressed genes and miRNAs in bursa of Fabricius in two breeds of duck[D].Ya'an:Sichuan Agricultural Uniersity,2015.(in Chinese)
[13] SETO F.Early development of the avian immune system[J].Poult Sci,1981,60(9):1981-1995.
[14] 方静,崔恒敏,廖婷彬,等.胚胎及胚后发育期鸭腔上囊淋巴细胞增殖与凋亡的研究[J].畜牧兽医学报,2008,39(11):1581-1587.
FANG J,CUI H M,LIAO T B,et al.Study on lymphocyte proliferation and apoptosis in the embryonic and post embryonic development of the duck bursa of Fabricius[J].Acta Veterinaria et Zootechnica Sinica,2008,39(11):1581-1587.(in Chinese)
[15] JIE H,LIAN L,QU L J,et al.Differential expression of Toll-like receptor genes in lymphoid tissues between Marek's disease virus-infected and noninfected chickens[J].Poult Sci,2013,92(3):645-654.
[16] 侯巍,岳华,汤承.荧光定量RT-PCR检测雏鸡感染NDV强毒后胸腺和法氏囊中TLR2与TLR4 mRNA的表达[J].中国预防兽医学报,2010,32(4):254-258.
HOU W,YUE H,TANG C.Development of a real-time RT-PCR for detection of Toll-like receptor2 and Toll-like receptor4 mRNAs in bursals and thymus of chicken infected with virulent NDV[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(4):254-258.(in Chinese)
[17] EFSTRATIADIS A,POSAKONY J W,MANIATIS T,et al.The structure and evolution of the human β-globin gene family[J].Cell,1980,21(3):653-668.
[18] 李萌.脊椎动物MAPK家族的进化研究和三角河蚌防御素基因(Defensins)的克隆与分析[D].镇江:江苏大学,2012.
LI M.The evolution of vertebrate MAPK family and the Triangle Mussel defensin gene (Defensins) cloning and analysis[D].Zhenjiang:Jiangsu University,2012.(in Chinese)
[19] DARFOUR-ODURO K A,MEGENS H J,ROCA A L,et al.Adaptive evolution of toll-like receptors (TLRs) in the family suidae[J].PLoS One,2015,10(4):e124069.
[20] LOPES-MARQUES M,CUNHA I,REIS-HENRIQUES M A,et al.Diversity and history of the long-chain acyl-CoA synthetase (Acsl) gene family in vertebrates[J].BMC Evol Biol,2013,13:271.
[21] WANG M H,ZHANG X Z,ZHAO H B,et al.FoxO gene family evolution in vertebrates[J].BMC Evol Biol,2009,9:222.
[22] ZHAO M,YING X X,SUN J,et al.Molecular evolution of Coq1 gene family in eukaryotes[J].J Syst Evol,2017,55(5):417-425.
[23] LAN D L,LIN B S,XIONG X R,et al.Identification and characteristics analysis of toll-like receptors family genes in yak[J].Genes Genomics,2016,38(5):429-438.
[24] 赵霞,张瑞莉,王海彬,等.IBDV感染SPF雏鸡法氏囊中TLR3表达的动态变化[J].黑龙江畜牧兽医,2016(5):168-170.
ZHAO X,ZHANG R L,WANG H B,et al.Dynamic changes in the expression of TLR3 in the bursa of Fabricius of chicks infected with IBDV[J].Heilongjiang Animal Science and Veterinary Medicine,2016(5):168-170.(in Chinese)
[25] 王建琳,曹志伟,王冬冬,等.不同致病性H9N2亚型禽流感病毒诱导鸡TLR-7和Mx基因mRNA转录水平的差异[J].畜牧兽医学报,2017,48(5):907-913.
WANG J L,CAO Z W,WANG D D,et al.Transcriptional responses of TLR-7 and Mx mRNA in 3 chicken host systems infected with H9N2 influenza viruses with different pathogenicities[J].Acta Veterinaria et Zootechnica Sinica,2017,48(5):907-913.(in Chinese)
[26] ZHANG A G,XU J H,LAI H Z,et al.Age-related changes and distribution of T cell markers (CD3 and CD4) and toll-like receptors(TLR2,TLR3,TLR4 and TLR7) in the duck lymphoid organs[J].Immunobiology,2017,222(7):857-864.
[27] LIU X D,ZHANG F B,SHAN H,et al.mRNA expression in different developmental stages of the chicken bursa of Fabricius[J].Poult Sci,2016,95(8):1787-1794.
[28] HASHIMOTO C,HUDSON K L,ANDERSON K V.The Toll gene of Drosophila,required for dorsal-ventral embryonic polarity,appears to encode a transmembrane protein[J].Cell,1988,52(2):269-279.
[29] MEADE K G,HIGGS R,LLOYD A T,et al.Differential antimicrobial peptide gene expression patterns during early chicken embryological development[J].Dev Comp Immunol,2009,33(4):516-524.
[30] 杭柏林,桑建君,钱琨,等.利用基因芯片分析鸡法氏囊差异表达基因[J].畜牧兽医学报,2014,45(7):1162-1169.
HANG B L,SANG J J,QIAN Q,et al.Analysis of differential expression genes in chicken bursa of Fabricius with gene expression array[J].Acta Veterinaria et Zootechnica Sinica,2014,45(7):1162-1169.(in Chinese)
[31] FULUI A,INOUE N,MATSUMOTO M,et al.Molecular cloning and functional characterization of chicken toll-like receptors.A single chicken toll covers multiple molecular patterns[J].J Biol Chem,2001,276(50):47143-47149.
[32] TIRUMURUGAAN K G,DHANASEKARAN S,RAJ G D,et al.Differential expression of toll-like receptor mRNA in selected tissues of goat (Capra hircus)[J].Vet Immunol Immunopathol,2010,133(2-4):296-301.
[33] TEMPERLEY N D,BERLIN S,PATON I R,et al.Evolution of the chicken toll-like receptor gene family:a story of gene gain and gene loss[J].BMC Genomics,2008,9:62.
[34] 汤龙军,李鹏程,朱璐,等.天冬氨酸蛋白酶在拟南芥和水稻中的分子进化、表达模式以及在花药发育中的功能分析[J].植物生理学报,2015,51(3):323-336.
TANG L J,LI P C,ZHU L,et al.Identification,evolutionary and expression profile analysis of the aspartic protease gene superfamily in arabidopsis thaliana and rice[J].Plant Physiology Journal,2015,51(3):323-336.(in Chinese)