临床疑似猪非典型瘟病毒感染仔猪小脑组织的转录差异  

Transcriptional differences in cerebellar tissue of clinically suspected atypical pestivirus infected piglets

作　　者：李雨濛 杨晓宇 朱玲 LI Yu-Meng;YANG Xiao-Yu;ZHU Ling(Animal Biotechnology Center,College of Veterinary Medicine,Sichuan Agricultural University,Chengdu,Sichuan 611130,China;Sichuan Key Laboratory of Animal Diseases and Human Health,Chengdu,Sichuan 611130,China)

机构地区：[1]四川农业大学动物医学院动物生物技术中心,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《微生物学通报》2020年第12期4122-4130,共9页Microbiology China

基　　金：四川省重大科技专项(2018NZDZX0006)。

摘　　要：【背景】猪非典型瘟病毒作为一种新发病毒,近几年在国内各省份中陆续被检出,目前针对猪非典型瘟病毒(atypical porcine pestivirus,APPV)的研究报道大多集中于基因组学和流行病学,而其感染宿主后的致病机制尚不清楚。【目的】利用高通量测序技术,对感染仔猪小脑组织进行转录水平差异研究,初步探索APPV致病机制。【方法】将实验组3份排除其余能引发仔猪先天性震颤和神经症状相关病原干扰的APPV感染仔猪的小脑组织,与对照组3份健康仔猪的小脑组织进行高通量测序,比较实验组和对照组之间转录水平的差异。【结果】构建了两个cDNA文库:APPV感染小脑组织文库、健康小脑组织文库。在P≤0.05,|Log2(FC)|≥0.263的筛选标准下,共筛选出差异表达基因(differentially expressed gene,DEG)381个,其中上调基因163个,下调基因218个;随机选取8个差异表达的基因,经RT-qPCR检测,以β-actin作为内参基因,结果显示8个差异基因的表达趋势与高通量测序一致。【结论】通过对临床感染APPV的仔猪小脑样本进行转录组研究,通过GO、KEGG对差异基因进行功能注释、分类,发现Pak4、Robo4、Sema3f、Wnt5a等参与轴突导向信号通路的基因转录水平显著下降,可能表明APPV感染可能会导致仔猪体内轴突导向过程受到抑制,进而使其中枢神经系统发育受到抑制,为APPV的致病机制等进一步研究提供了方向和指导。[Background]Atypical porcine pestivirus(APPV)as a new virus,it has been detected in various provinces in China in recent years.At present,most of the research reports on APPV focus on genomics and epidemiology.The pathogenic mechanism after infection of the host is unclear.[Objective]To investigate the pathogenicity of APPV in the cerebellum of infected piglets by high-throughput sequencing technology.[Methods]Three cerebellum of APPV-infected piglets of experimental group excluded from pathogens which can cause congenial tremors(CT)or neurological symptoms were sequenced with 3 cerebellum of healthy piglets of control group,and compare the differences in transcription levels between experimental and control groups.[Results]Two cDNA libraries were constructed in this study:APPV infected cerebellum tissue library and healthy cerebellum tissue library.Under the screening criteria of P≤0.05,|Log2(FC)|≥0.263,a total of 381 differentially expressed genes(DEGs)were screened,including 163 up-regulated genes and 218 down-regulated genes;8 differentially expressed genes were randomly selected.The gene was detected by RT-qPCR andβ-actin was used as an internal reference gene.The expression trends of eight differential genes were consistent with high-throughput sequencing.[Conclusion]A transcriptome study was conducted on the cerebellum samples of clinically infected piglets with APPV.Based on the GO and KEGG,we performed the functional annotation and classification of DEGs.It was found that the transcriptional levels of Pak4,Robo4,Sema3 f,Wnt5 a and other genes involved in axon guidance signaling pathways had significantly decreased.This result indicates that APPV infection may inhibit the axon guidance process in piglets.Furthermore,it inhibits the development of the central nervous system,which provides direction and guidance for further research on the pathogenic mechanism of APPV.

关 键 词：猪非典型瘟病毒 转录组学 差异表达基因 轴突导向 

分 类 号：S858.28[农业科学—临床兽医学]