不同滴度鸭肠炎病毒感染对鸭脾脏的转录组变化分析
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摘要
为探讨不同滴度鸭肠炎病毒(duck enteritis virus,DEV)感染对鸭组织器官转录组的影响,本研究采用2个不同DELD50滴度的DEV,接种50日龄健康鸭90h后,采集脾脏样本,高通量测序技术进行转录组测序,筛选差异表达基因,并应用GO与KEGG数据库进行分析。结果显示,当接种量为100×DELD5 0时,鸭脾脏差异表达基因有685个,其中上调基因341个,主要参与免疫反应、核糖体结构和酶活性等生物学过程,并在核糖体、氧化磷酸化和吞噬体信号通路中显著富集;下调基因为344个,主要参与细胞分化正调控、β转化生长因子生成正调控、酶与相关受体活性、细胞结构等生物学过程,并在细胞黏附分子、内吞作用、肠道免疫网络IgA产生、ECM受体互作、缝隙连接和黏着信号通路上显著富集。当接种量为10-2×DELD50时,鸭脾脏差异表达基因有485个,其中上调基因297个,主要参与细胞功能、蛋白结合和蛋白复合物等生物学过程,并在细胞黏附分子、吞噬体、肠道免疫网络IgA产生、球系列鞘糖脂生物合成及N-糖链合成信号通路中显著富集;下调基因188个,主要参与补体激活、肌肉组织活动调节、受体复合物、酶与受体活性等生物学过程,并在基础转录因子、PPAR信号通路、α-亚麻酸代谢等代谢途径与信号通路上显著富集。这些结果表明不同滴度DEV感染对鸭脾脏转录组产生不同的影响,为深入探究DEV致病分子机制提供基础资料。
To investigate the effects of duck enteritis virus(DEV)infection with different titers on the transcriptome of duck tissue,two LD50 titers of DEV were used to inoculate the 50 day old healthy ducks,and the spleen samples of ducks were collected at 90 hinoculation for the transcriptome sequencing by high-throughput technology and the differentially expressed genes were analyzed by GO and KEGG bioinformatics.The results showed as the follows:when the inoculation amount was 100×DELD50 of DEV,there were 685 of differentially expressed genes in duck spleen,among which 341 were up-regulated genes,mainly involved in immune response,ribosome structure and enzyme activity and other biological processes,which significantly enriched in the ribosome,oxidative phosphorylation and phagosome signal pathways.And 344 were down-regulated genes,mainly involved in positive regulation of cell differentiation and transforming growth factor beta production,enzyme and receptor activity,cell structure and other biological processes,which significantly enriched in cell adhesion molecules,endocytosis,intestinal immune network for IgA production,ECM receptor interaction,gap junction and focal adhesion signal pathways.When the inoculation amount was 10-2×DELD50 of DEV,there were 485 of differentially expressed genes in duck spleen,among which 297 were up-regulated genes,mainly involved in cell function,protein binding,protein complexes and other biological processes,which significantly enriched in phagosome,cell adhesion molecules,intestinal immune network for IgA production,glycosphingolipid biosynthesis-globo series and N-glycan biosynthesis.And 188 were down-regulated genes,mainly involved in complement activation,muscle activity regulation,receptor complexes,enzyme and receptor activity and other biological processes,which significantly enriched in basal transcription factors,PPAR signaling pathway and alpha linolenic acid metabolism.These results indicated that the DEV infection with different titers could be a different influence on duck spleen transcriptome,which provide the basic data for exploring the molecular mechanism of DEV infection.
To investigate the effects of duck enteritis virus(DEV)infection with different titers on the transcriptome of duck tissue,two LD50 titers of DEV were used to inoculate the 50 day old healthy ducks,and the spleen samples of ducks were collected at 90 hinoculation for the transcriptome sequencing by high-throughput technology and the differentially expressed genes were analyzed by GO and KEGG bioinformatics.The results showed as the follows:when the inoculation amount was 100×DELD50 of DEV,there were 685 of differentially expressed genes in duck spleen,among which 341 were up-regulated genes,mainly involved in immune response,ribosome structure and enzyme activity and other biological processes,which significantly enriched in the ribosome,oxidative phosphorylation and phagosome signal pathways.And 344 were down-regulated genes,mainly involved in positive regulation of cell differentiation and transforming growth factor beta production,enzyme and receptor activity,cell structure and other biological processes,which significantly enriched in cell adhesion molecules,endocytosis,intestinal immune network for IgA production,ECM receptor interaction,gap junction and focal adhesion signal pathways.When the inoculation amount was 10-2×DELD50 of DEV,there were 485 of differentially expressed genes in duck spleen,among which 297 were up-regulated genes,mainly involved in cell function,protein binding,protein complexes and other biological processes,which significantly enriched in phagosome,cell adhesion molecules,intestinal immune network for IgA production,glycosphingolipid biosynthesis-globo series and N-glycan biosynthesis.And 188 were down-regulated genes,mainly involved in complement activation,muscle activity regulation,receptor complexes,enzyme and receptor activity and other biological processes,which significantly enriched in basal transcription factors,PPAR signaling pathway and alpha linolenic acid metabolism.These results indicated that the DEV infection with different titers could be a different influence on duck spleen transcriptome,which provide the basic data for exploring the molecular mechanism of DEV infection.
引文
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[16]杨晓燕,程安春,汪铭书,等.鸭瘟病毒强毒株在感染鸭实质器官内的增殖与分布[J].中国兽医学报,2008,28(11):1254-1258.
[17]韩晓英.人工感染鸭瘟病毒急性病理模型的病理学发展规律及免疫荧光检测石蜡切片中鸭瘟病毒分布规律的研究[D].四川雅安:四川农业大学,2004.
[18]张坤,刁有祥,程彦丽,等.鸭瘟病毒强毒株感染SPF鸭动态病理组织学变化[J].中国兽医学报,2013,33(1):9-10.
[19]齐雪峰,罗薇,杨晓燕,等.鸭瘟病理组织学动态观察[J].中国预防兽医学报,2006,28(1):44-47.
[20]HE L,XUE R Y,CAO G L,et al.Analysis of highly expressed genes in Bombyx mori ovarian culture cells(BmN)based on RNA-Seq technology[J].Sci Sericul,2014,40(3):409-421.
[21]JIA X P,YE X Q,LIANG L J,et al.Transcriptome characteristics of Paspalum vaginatum analyzed with Illumina sequencing technology[J].Acta Prataculturae Sinica,2014,23(6):242-252.
[22]CHEN D,FU S Y,GAO H Y,et al.Transcriptome analysis of intranmuscular preadipocytes and matureadipocyte in cashmere goats[J].Acta Veterin Zootech Sinica,2014,45(5):714-721.
[23]DAUDE N.Prion diseases and the spleen[J].Viral Immunol,2004,17(3):334-349.
[2]陈溥言.兽医传染病学[M].5版.北京:中国农业出版社,2006:383-385.
[3]侯玉臻,赵丹彤,刘国英,等.基于高通量测序技术分析家兔感染猪瘟兔化弱毒株后脾脏的转录组学变化[J].病毒学报,2016,32(3):316-323.
[4]MOHAMMAD H,MARIANNE H,DMITRY K,et al.Transcriptional profiling of Mareks disease virus genes during cytolytic and latent infection[J].Virus Gen,2008,36(2):383-392.
[5]HUANG Y H,HUANG X H,YAN Y,et al.Transcriptome analysis of orange-spotted grouper(Epinephelus coioides)spleen in response to Singapore grouper iridovirus[J].BMC Genom,2011,12(1):556.
[6]JAE H P,HYUN D J,SUHEE H.Analysis of immune gene expression in rock bream spleen after rock bream iridovirus infection by transcriptome sequencing[J].Fish Shellfish Immunol,2016,53:96.
[7]NIE Q H,SANDTORD E E,ZHANG X Q,et al.Deep sequencing-based transcriptome analysis of chicken spleen in response to avian pathogenic Escherichia coli(APEC)infection[J].PLoS One,2012,7(7):e41645.
[8]YANG Y,YU H,LI H,et al.Transcriptome profiling of grass carp(Ctenopharyngodon idellus)infected with Aeromonas hydrophila[J].Fish Shellfish Immunol,2016,51:329-336.
[9]XU B H,ZHONG L,LIU Q L,et al.Characterization of grass carp spleen transcriptome during GCRV infection[J].Genet Mol Res,2016,15(2):6650.
[10]张英楠,杨树宝,杨柳,等.鸡脾脏组织结构早期发育的动态观察[J].中国家禽,2010,32(3):13-15.
[11]孙小光,欧阳军.脾脏的免疫功能及脾脏对血流变影响的研究[J].现代生物医学进展,2009,19(9):3776-3778.
[12]MORTAZAVI A,WILLIAMS B A,MCCUE K,et al.Mapping and quantifying mammalian transcriptomes by RNA-Seq[J].Nat Methods,2008,5(7):621-628.
[13]YUAN G P,CHENG A C,WANG M S,et al.Electron microscopic studies of the morphogenesis of duck enteritis virus[J].Avian Dis,2005,49(1):50-55.
[14]文晶亮,李俊平,李启红,等.鸭肠炎病毒强毒参考株在感染鸭体内的动态分布研究[J].中国兽药杂志,2014,48(12):10-16.
[15]刘菲,程安春,汪铭书,等.鸭瘟病毒强毒株在急性人工感染成年鸭病例体内分布规律[J].中国兽医学报,2004,24(3):228-230.
[16]杨晓燕,程安春,汪铭书,等.鸭瘟病毒强毒株在感染鸭实质器官内的增殖与分布[J].中国兽医学报,2008,28(11):1254-1258.
[17]韩晓英.人工感染鸭瘟病毒急性病理模型的病理学发展规律及免疫荧光检测石蜡切片中鸭瘟病毒分布规律的研究[D].四川雅安:四川农业大学,2004.
[18]张坤,刁有祥,程彦丽,等.鸭瘟病毒强毒株感染SPF鸭动态病理组织学变化[J].中国兽医学报,2013,33(1):9-10.
[19]齐雪峰,罗薇,杨晓燕,等.鸭瘟病理组织学动态观察[J].中国预防兽医学报,2006,28(1):44-47.
[20]HE L,XUE R Y,CAO G L,et al.Analysis of highly expressed genes in Bombyx mori ovarian culture cells(BmN)based on RNA-Seq technology[J].Sci Sericul,2014,40(3):409-421.
[21]JIA X P,YE X Q,LIANG L J,et al.Transcriptome characteristics of Paspalum vaginatum analyzed with Illumina sequencing technology[J].Acta Prataculturae Sinica,2014,23(6):242-252.
[22]CHEN D,FU S Y,GAO H Y,et al.Transcriptome analysis of intranmuscular preadipocytes and matureadipocyte in cashmere goats[J].Acta Veterin Zootech Sinica,2014,45(5):714-721.
[23]DAUDE N.Prion diseases and the spleen[J].Viral Immunol,2004,17(3):334-349.