基于转录组测序数据的山羊下丘脑组织繁殖相关信号通路及候选基因的筛选  

Selection of Related Signaling Pathways and Candidate Genes Related to Hypothalamus Tissue Reproduction in Goats Based on Transcriptome Sequencing Data

作　　者：施会彬 王玉琴[1,2] 张小辉 吴至博 杨芳 SHI Huibin;WANG Yuqin;ZHANG Xiaohui;WU Zhibo;YANG Fang(Henan University of Science and Technology,Luoyang 471023,China;Henan ProvinceMutton Sheep Breeding Engineering Technology Research Center,Luoyang 471023,China)

机构地区：[1]河南科技大学,洛阳471023 [2]河南省肉羊繁育工程技术研究中心,洛阳471023

出　　处：《畜牧兽医学报》2019年第7期1380-1395,共16页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金面上课题(31472095)

摘　　要：旨在通过对不同光照时长条件下河南槐山羊下丘脑组织进行转录组测序和生物信息学分析,筛选出与山羊繁殖相关的信号通路及候选基因。本研究利用Illumina二代高通量测序平台(NGST),采用PE150测序策略,对经自然光照(8 h光照:16 h黑暗)与人工光照(16 h光照:8 h黑暗)条件处理后的8只空怀母羊(每组各4只,平均年龄1周岁)下丘脑组织进行转录组测序,将组装得到的Unigene总数比对到参考基因组序列,进行差异表达基因分析、GO和KEGG富集分析以及候选基因筛选,并采用Real-time PCR方法对候选基因的相对表达量变化进行分析。结果显示,RNA-Seq共得到了约4.4亿条reads,平均每个样本的reads数约为5 249万条;自然光照与人工光照2组DESeq分析得到448个差异表达基因,并富集在KEGG数据库中的241个信号通路,其中包括5个与繁殖相关的信号通路;3个速激肽家族候选基因(TACR1、TACR2和TACR3)显著富集在Calcium signaling pathway(钙离子信号通路,chx04020)通路;Real-time PCR分析结果显示,转录组测序结果可靠。综上表明,Calcium signaling pathway(钙离子信号通路,chx04020)与TACR1、TACR2和TACR3基因,可能在山羊繁殖过程中发挥着重要作用。The aim of this study was to screen the signaling pathways and candidate genes related to goat reproduction of hypothalamus tissue of Henan Huai goats under different illumination conditions by transcriptome sequencing and bioinformatics analysis. The transcriptome sequencing of 8 infecund ewes hypothalamus tissue(4 in each group, one year old) treated with natural light(8 h light: 16 h dark) and artificial light(16 h light: 8 h dark) were performed by using a second generation high throughput sequencing platform(NGST), and PE150 sequencing strategy. The total number of unigene assembled was compared to the reference genome sequence, differentially expressed genes(DEGs) analysis, Go and KEGG enrichment analysis and candidate gene screening were performed. The relative expression of candidate genes was analyzed by Real-time PCR. The results showed that about 440 million reads were obtained by RNA-Seq, and the average reads number per sample was about 52.49 million. Four hundred and forty-eight differentially expressed genes were obtained by DESeq analysis between natural and artificial illumination groups, which were enriched in 241 signaling pathways in KEGG database, including 5 reproductive-related signaling pathways. Three candidate genes of tachykinin family(TACR1,TACR2 and TACR3) were significantly enriched in the calcium signaling pathway(chx04020). The results of Real-time PCR analysis showed that the sequencing results of the transcriptome were reliable. These results suggest that calcium signaling pathway(chx04020) and TACR1,TACR2 and TACR3 genes may play the important role in goat reproduction.

关 键 词：河南槐山羊 下丘脑 转录组 信号通路 差异表达基因 速激肽 

分 类 号：S826.2[农业科学—畜牧学]