摘要
旨在筛选与季节性发情相关的核心基因和长链非编码RNA(long non-coding RNA,lncRNA)。本研究选取2~3岁空怀的中国美利奴羊和湖羊母羊(乏情期、发情期和发情间期各3只),采集卵巢组织样本,采用RNA-seq技术筛选差异表达的基因(differentially expressed genes,DEGs)和lncRNAs,并通过GO、KEGG富集分析、PPI分析及ceRNA网络构建鉴定与繁殖性能相关的核心基因和lncRNAs。结果,构建了6个不同生理状态下的RNA文库,筛选得到1495个差异表达基因(differentially expressed genes,DEGs)和454个差异表达lncRNAs,其中共性DEGs和lncRNAs分别为313个和435个,特有DEGs和lncRNAs分别为1182个和19个。GO和KEGG富集分析显示,共性DEGs在细胞分裂和细胞周期等方面显著富集,而差异表达的lncRNAs靶基因富集于细胞内信号转导及负调控RNA聚合酶II启动子转录等。特有DEGs在细胞迁移的正向调节中富集,lncRNAs的靶基因集中于细胞间粘附和整合素介导的信号通路等。通过PPI筛选出19个核心基因,主要富集在卵母细胞减数分裂和细胞周期信号通路。聚类分析结果显示,核心基因在乏情期表达量较高。ceRNA网络进一步揭示基因在细胞周期等信号通路的富集。RT-qPCR验证结果与高通量测序一致。筛选出19个核心基因和28个lncRNAs,并发现42个lncRNA-miRNA-mRNA靶向关系,为理解绵羊季节性繁殖的分子机制提供了潜在调控靶点。
The aim of this study was to screen core genes and long non-coding RNAs related to seasonal estrus.Ovarian tissue were collected from 2 to 3 years old,non-pregnant female Chinese Merino and Hu sheep(3 sheep per group,each in anestrus,estrus and interestrus).RNA-seq technology was employed to identify differentially expressed genes and lncRNAs.GO and KEGG enrichment analyses,protein-protein interaction(PPI)network analysis and ceRNA network construction were performed to identify core genes and lncRNAs related to reproductive performance.RNA libraries for 6 distinct physiological states were constructed.There were 1495 differentially expressed genes(DEGs)and 454 differentially expressed long non-coding RNAs(lncRNAs)be identified.Among these,313 DEGs and 435 lncRNAs were common,while 1182 DEGs and 19 lncRNAs were unique.GO and KEGG enrichment analyses revealed that common DEGs were significantly enriched in the processes related to cell division and the cell cycle,whereas the target genes of differentially expressed lncRNAs were enriched in intracellular signaling pathways and negative regulation of RNA polymerase II promoter transcription.Unique DEGs were primarily associated with the positive regulation of cell migration.The target genes of lncRNAs were enriched in cell adhesion and integrin-mediated signaling pathways.There were 19 core genes identified by protein-protein interaction network analysis,which were predominantly enriched in oocyte meiosis and cell cycle pathways.These core genes exhibited higher expression level during the anestrus phase than those in interestrus.Further analysis of the ceRNA network highlighted the enrichment of genes in cell cycle-related signaling pathways.RT-qPCR results were consisted with the RNA-seq results.A total of 19 core genes and 28 lncRNAs were identified,along with 42 lncRNA-miRNA-mRNA interactions,providing potential regulatory targets for understanding the molecular mechanisms of seasonal reproduction in sheep.
作者
杨杨
李良远
万鹏程
卢守亮
刘长彬
杨华
王立民
代蓉
周平
YANG Yang;LI Liangyuan;WAN Pengcheng;LU Shouliang;LIU Changbin;YANG Hua;WANG Limin;DAI Rong;ZHOU Ping(State Key Laboratory of Sheep Genetic Improvement and Healthy Production,Xinjiang Academy of Agricultural and Reclamation Science,Shihezi 832000,China;Institute of Traditional Chinese Medicine and Ethnical Medicine Resources,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)
出处
《畜牧兽医学报》
北大核心
2025年第3期1264-1277,共14页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
兵团重点领域科技攻关项目
兵团农业创新工程(NCG202221)
中央引导地方科技发展资金项目
省部共建绵羊遗传改良与健康养殖国家重点实验室建设(2022CA002)。
作者简介
杨杨(1989-),女,河南西平人,硕士,主要从事动物分子遗传与育种研究,E-mail:1010975408@qq.com;通信作者:代蓉,主要从事动物分子遗传与育种研究,E-mail:dairong1@163.com;通信作者:周平,主要从事动物分子遗传与育种研究,E-mail:zhpxqf@163.com。
