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基于转录组测序分析牛不同脂肪组织的脂肪沉积差异研究
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引用本文:王思元,刘 迪,张伟红,王国富,高树新.基于转录组测序分析牛不同脂肪组织的脂肪沉积差异研究[J].西北农业学报,2021,(12):1755~1766
DOI:10.7606/j.issn.1004-1389.2021.12.001
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作者单位
王思元,刘 迪,张伟红,王国富,高树新 (1.内蒙古民族大学 动物科技学院内蒙古通辽 0280002. 内蒙古通辽市开鲁县兽医局内蒙古通辽 028000) 
基金项目:内蒙古自然科学基金(2019MS03077); 内蒙古自治区第五批"草原英才"产业创新创业人才团队"肉牛科学研究与肉牛产业创新人才团队"专项;内蒙古重大专项"科尔沁肉牛品种培育"(2060901); 内蒙古自治区科技计划项目"科尔沁肉牛新品种选育集成技术研究与示范推广"(KJXM2020002-05)。
中文摘要:为了更好地了解肉牛(Bos taurus)的脂肪沉积机制,以安格斯牛(Aberdeen Angus)和西门塔尔牛(Simmental cattle)为模型,通过对2种脂肪组织(肾周脂肪和背皮下脂肪)进行RNA测序,确定转录组图谱。分析4个组织中共同的高表达基因,以及2个组织之间的差异表达基因。结果显示,每个样品获得约10.99 Gb数据,并注释总共23 472个基因。在50个高度表达的共同基因中,发现脂肪酸结合蛋白4(Fatty Acid Binding Protein4, FABP4),脂肪形成调节因子(Adipogenesis Regulatory Factor,ADIRF)和硬脂酰辅酶A去饱和酶(Stearoyl-CoA Desaturase,SCD)是与脂肪沉积相关的基因,表明这3个基因可能在脂肪沉积中发挥作用。在安格斯牛和西门塔尔牛的肾周脂肪与皮下脂肪组织之间分别鉴定出1 678个和1 955个差异表达基因。GO分析表明,一些DEG与代谢有关。KEGG途径分析显示,PPAR信号途径、甘油酯代谢和ECM-受体相互作用途径富集丰富。在背皮下脂肪中,PPAR信号途径和甘油脂代谢中的3个基因:视黄酸X受体α(retinoid X receptor alpha,RXRA)、C1q肿瘤坏死因子相关蛋白7(C1q and TNF related 7, C1QTNF7)和单酰甘油-O-酰基转移酶2(Monoacylglycerol O-acyltransferase 2, MOGAT2)上调;6个基因:肉碱脂酰转移酶1B(Carnitine Palmitoyltransferase 1B, CPT1B) 、解偶联蛋白1(Uncoupling Protein 1, UCP1)、 脂肪酸结合蛋白3(Fatty Acid Binding Protein 3, FABP3)、 脂肪酸转位酶(Fatty Acid Translocation enzyme, CD36) 、脂蛋白1( LPIN1) 和甘油-3-磷酸酰基转移酶3(Glycerol-3-phosphate Acyltransferase 3, GPAT3)表达量下调。在ECM-受体相互作用中,发现Ⅰ型胶原Α1(Collagen Type I alpha 1 Chain, COL1A1)、III型胶原Α1(Collagen Type III alpha 1 Chain, COL3A1) 、Ⅰ型胶原Α2(Collagen Type I alpha 2 Chain, COL1A2)和II型胶原Α1(Collagen Type II alpha 1 Chain, COL2A1)在牛的背皮下脂肪中上调,而层粘连蛋白γ3(Laminin Subunit gamma 3 , LAMC3) 和粘连蛋白γ2(Laminin Subunit gamma 2, LAMC2)的表达量下调。为验证转录组的准确性,对差异基因 COL1A1 COL3A1RXRA LPIN1 GPAT3进行qPCR验证,结果显示qPCR结果与转录组测序数据基本一致。本研究揭示了肉牛不同脂肪组织中复杂的转录组图谱,发掘了参与脂肪沉积的候选基因。
中文关键词:  脂肪组织  脂肪沉积  转录组测序  候选基因
 
Analysis of the Difference of Fat Deposition in Different Adipose Tissues of Cattle Based on Transcriptome Sequencing
Abstract:In order to explore the biological difference between perirenal visceral fat and back subcutaneous fat,mRNA from the fat tissue of Angus cattle and Simmental cattle were extracted,and RNA-seq was done to identify the common highly expressed genes in four tissues and the differentially expressed genes (DEGs) between the perirenal visceral fat and back subcutaneous fat. The results showed that 10.99 Gb data were generated in each library and 23 472 genes were identified. Three genes including fatty acid binding protein4( FABP4),adipogenesis regulatory factor (ADIRF) and stearoyl-CoA desaturase (SCD) that involved in fat depot were common highly expressed in the four tissues. There were 1 678 DEGs between perirenal visceral fat and subcutaneous fat in Angus cattle and 1 955 DEGs in Simmental cattle. Several DEGs involving in metabolism was identified by GO function analysis. PPAR signal pathway,glycerolipid metabolism and ECM-receptor interaction pathway were significantly enriched. Retinoid X receptor alpha (RXRA),C1q, TNF related 7 ( C1QTNF7) and monoacylglycerol O-acyltransferase 2 ( MOGAT2) that played a key role in PPAR signal pathway and glycerolipid metabolism were upregulated in subcutaneous fat,while carnitine palmitoyltransferase 1B ( CPT1B),uncoupling protein 1 ( UCP1),fatty acid binding protein 3 ( FABP3),fatty acid translocation enzyme ( CD36),glycerol-3-phosphate acyltransferase 3 ( GPAT3),LPIN1 were down regulated. Collagen type I alpha 1 chain ( COL1A1),Collagen type III alpha 1 chain ( COL3A1),Collagen type I alpha 2 chain ( COL1A2) and Collagen type II alpha 1 chain ( COL2A1) related to ECM-receptor interaction pathway were upregulated in subcutaneous fat,while laminin subunit gamma 3 ( LAMC3) and Laminin Subunit gamma 2 ( LAMC2) were down regulated. The qRT-PCR assays were conducted to validate five selected DEGs from RNA-Seq: COL1A1 COL3A1 RXRA LPIN1 and GPAT3. The relative expression changes of qRT-PCR data were correlated with RNA-Seq data. The present study explored the difference of mRNA profile between different adipose tissues,and  identified the candidate genes that contributed to fat depot,which will provide a foundation for further experiment on the role of these genes in the regulation of fat depot.
keywords:Cattle  Adipose tissue  Adipose deposition  Transcriptome sequencing  Candidate genes
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