基于转录组测序挖掘犊牦牛肾周脂质沉积关键基因

Exploring Candidate Genes Related to Kidney-adjacent Lipid Deposition in Yak Calves Based on Transcriptome Sequencing

【目的】比较不同日龄犊牦牛肾周脂肪组织转录组表达谱的变化,挖掘调控犊牦牛肾周脂质沉积的信号通路和关键基因。【方法】选取1、7和30日龄各3头健康雄性犊牦牛,放血处死,采集肾周脂肪组织,利用Illumina HiSeq平台进行转录组测序,对转录组数据进行质控、比对、差异表达基因(DEGs)筛选,并对DEGs进行GO功能和KEGG通路富集分析。随机选取7个基因进行实时荧光定量PCR验证。【结果】转录组结果显示,在1和7日龄比较组中共鉴定到1627个显著DEGs,其中939个上调,688个下调;在7和30日龄比较组中共鉴定到3979个显著DEGs,其中1925个上调,2054个下调。GO功能和KEGG通路富集分析结果显示,在1和7日龄比较组中DEGs富集到酰基辅酶A脱氢酶活性条目,参与了脂肪酸生物合成、胰岛素抵抗、PPAR信号通路及鞘磷脂信号通路等;在7和30日龄比较组中DEGs富集到脂肪酸代谢、脂质代谢、磷酸水解酶活性及脂肪酶活性等多个生物过程,参与了丁酸盐代谢、脂肪酸代谢、甘油磷脂代谢、磷脂酶D信号通路及AMPK信号通路等。从富集通路中发现,SCD、ADIPOQ、PLIN1、ACADM和LPL等基因是与犊牦牛肾周脂质沉积相关的候选基因。实时荧光定量结果显示,7个基因的表达与转录组测序结果一致。【结论】本研究筛选出SCD、ADIPOQ、PLIN1、ACADM和LPL共5个与脂肪细胞分化和脂质沉积相关的基因。研究结果为进一步探究不同生长阶段犊牦牛肾周脂质沉积提供理论依据。

【Objective】This study was aimed to compare the changes in transcriptome expression profiles of kidney-adjacent adipose tissue in yak calves at different ages,and explore the signaling pathways and key genes that regulated kidney-adjacent lipid deposition in yak calves.【Method】Three healthy male yak calves at 1,7 and 30 day-old were bled to death,and the kidney-adjacent adipose tissue were collected for transcriptome sequencing using Illumina HiSeq platform,after which the transcriptome data were subjected to quality control,comparison,differentially expressed gene(DEGs)screening,and the DEGs were performed to GO function and KEGG pathway enrichment analysis.In addition,seven genes were randomly selected for Real-time quantitative PCR verification.【Result】Transcriptome results showed that a total of 1627 significantly DEGs were identified in the comparison group of 1-and 7-day-old,of which 939 were up-regulated and 688 were down-regulated.Also,a total of 3979 significantly DEGs were identified in the comparison group of 7-and 30-day-old,of which 1925 were up-regulated and 2054 were down-regulated.GO function and KEGG pathway enrichment analysis showed that the DEGs were enriched in acyl-CoA dehydrogenase activity to participate in fatty acid biosynthesis,insulin resistance,PPAR signaling pathway,sphingolipid signaling pathway,etc.in the comparison group of 1-and 7-day-old.In addition,the DEGs were enriched in several biological processes,including fatty acid metabolism,lipid metabolism,phosphohydrolase activity and lipase activity to participate in butyrate metabolism,fatty acid metabolism,glycerophospholipid metabolism,phospholipase D signaling pathway,AMPK signaling pathway,etc.in the comparison group of 7-and 30-day-old.The enriched pathways showed that SCD,ADIPOQ,PLIN 1,ACADM and LPL were identified as candidate genes associated with kidney-adjacent lipid deposition in yak calves.Real-time quantitative PCR results showed that the expression of seven genes were consistent with RNA-Seq results.【Conclusion】Five genes related to adipocyte differentiation and fat deposition were screened in this study,including SCD,ADIPOQ,PLIN 1,ACADM and LPL.The results provided a theoretical reference for further research on kidney-adjacent lipid deposition in yak calves at different growth stages.