基于RNA-Seq技术的山羊嘴唇黏膜色素沉积相关差异表达基因筛选及初步分析  

Screening and preliminary analysis of differentially expressed genes related to pigmentation in goat lip mucosa based on RNA-Seq technology

作　　者：蒋婧[1,2] 屈明好 孙晓燕 李杰[1,2] 陈灿灿 李世勇[3] 任航行 JIANG Jjng;QU Minghao;SUN Xiao-yan;LI Jie;CHEN Can-can;LI Shi-yong;REN Hang-xing(Chongqing Academy of Animal Sciences,Rongchang,Chongqing 402460,China;Chongqing Engineering Research Center for Goat,Rongchang,Chongqing 402460,China;Fengming Town People's Government,Yunyang,Chongqing 404500,China)

机构地区：[1]重庆市畜牧科学院,重庆荣昌402460 [2]重庆市山羊工程技术研究中心,重庆荣昌402460 [3]云阳县凤鸣镇人民政府,重庆云阳404500

出　　处：《西南农业学报》2024年第6期1361-1369,共9页Southwest China Journal of Agricultural Sciences

基　　金：重庆市技术创新与应用发展专项重点项目(cstc2021jscx-gksbX0008);重庆市畜牧科学院市级财政科研项目(23511C、22544C)。

摘　　要：【目的】通过RNA-Seq技术筛选不同颜色山羊嘴唇色素代谢相关的候选基因,初步揭示山羊嘴唇黏膜颜色差异形成的分子机理,为动物黏膜色素代谢研究提供理论依据。【方法】以嘴唇等面部可视黏膜颜色存在差异的酉州乌羊(乌黑色)和板角山羊(粉色)为研究对象,采用Illumina Hiseq 2000测序平台对嘴唇黏膜组织进行转录组测序,所得序列经质控、组装后,以|log_(2)(Fold Change)|>1且q value<0.05为标准筛选差异表达基因,并对差异表达基因进行功能注释和富集分析。【结果】利用RNA-Seq技术共筛选出249个基因差异表达,其中与粉色嘴唇黏膜组织相比,乌黑色嘴唇黏膜组织中上调基因有218个,下调基因有31个。23个差异表达基因被划分在9种TFs家族中,其中14个基因属于含有锌指结构域的转录因子家族。GO功能富集分析结果显示,差异表达基因主要参与离子转运相关生物学过程和分子功能。KEGG代谢通路富集结果显示,差异表达基因显著富集于催产素、钙、cGMP-PKG信号通路,以及肥厚性心肌病、心脏肌肉收缩等7个与肌肉组织功能相关的信号通路,其中钙和cGMP-PKG信号通路与色素代谢密切相关;此外,ASIP等12个基因富集到MAPK信号通路、cAMP信号通路、黑色素合成信号通路(Melanogenesis)等色素代谢核心通路中;CALML6、OXTR、ATP1A2、ATP1B44个差异表达基因同时富集到多条与色素代谢相关的通路。qRT-PCR结果与测序结果一致,表明测序结果可靠。【结论】初步确定ASIP基因为山羊嘴唇色素代谢的候选基因,CALML6、OXTR、ATP1A2、ATP1B44个基因参与了山羊嘴唇黏膜色素沉积过程,为研究山羊黏膜色素代谢的分子机制提供理论基础,也为探究人类黏膜异常着色的分子机制提供参考。【Objective】The present study aimed to screen candidate genes related to the metabolism of lip pigment in goats with different colors screened by RNA-Seq technology,and the molecular mechanism of the color difference of goat lip mucosa was initially revealed,which provided theoretical basis for the study of animal mucosal pigment metabolism.【Method】The lip mucosa tissues of Youzhou Dark-skin goats(black)and Banjiao goats(pink)with different facial mucosal color were selected as the study objects.The transcriptome sequencing was performed by Illumina Hiseq 2000 sequencing platform.After quality control and assembly,the differentially expressed genes were screened using|log_(2)(Fold Change)|>1 and q value<0.05 as the criteria.Functional annotation and enrichment analysis of differentially expressed genes were performed.【Result】A total of 249 differentially expressed genes were screened by RNA-Seq technology.Compared with the pink mucosal tissues,218 up-regulated genes and 31 down-regulated genes were found in the black mucosal tissues.A total of 23 differentially expressed genes were predicted to be classified into 9 TFs families,of which 14 genes were classified into transcription factor families containing zinc finger domains.GO functional enrichment analysis showed that differentially expressed genes were mainly involved in various ion transport-related biological processes and molecular functions.The enrichment results of KEGG metabolic pathways showed that the differentially expressed genes were significantly enriched in oxytocin,calcium,cGMP-PKG signaling pathways,and other 7 signaling pathways related to muscle tissue function,include hypertrophic cardiomyopathy and cardiac muscle contraction,among which calcium and cGMP-PKG signaling pathways were closely related to pigment metabolism.In addition,12 genes including ASIP were enriched into the core metabolic pathways of pigment such as MAPK signaling pathway,cAMP signaling pathway,and Melanogenesis signaling pathway.Four differentially expressed g

关 键 词：山羊 转录组测序 嘴唇黏膜 色素沉积 差异表达基因 

分 类 号：S827[农业科学—畜牧学]