gga-miR-17-5p在鸡骨骼肌和脂肪沉积相关组织及细胞中的表达差异及其关键靶基因的筛选  

Differential expression of gga-miR-17-5p in chicken adipose and skeletal muscle tissues and cells and screening of its key target genes

作　　者：杨苗苗 向海 李瑞瑞 赵振华[1] 王钱保[1] 黄正洋[1] 李春苗[1] 梁忠[1] 吴兆林[1] 黄华云[1] YANG Miaomiao;XIANG Hai;LI Ruirui;ZHAO Zhenhua;WANG Qianbao;HUANG ZhengYang;LI Chunmiao;LIANG Zhong;WU Zhaolin;HUANG Huayun(Institute of Poultry Science,Yangzhou 225125,China;Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding/School of Life Science and Engineering,FOSHAN University,Foshan 528225,China)

机构地区：[1]江苏省家禽科学研究所,江苏扬州225125 [2]佛山科学技术学院生命科学与工程学院/广东省动物分子设计与精准育种重点实验室,广东佛山528225

出　　处：《中国农业大学学报》2024年第7期23-33,共11页Journal of China Agricultural University

基　　金：国家重点研发计划“农业生物种质资源挖掘与创新利用”重点专项资助(2021YFD1200803);现代农业产业技术体系建设专项(CARS-41-Z05);江苏省种业振兴揭榜挂帅项目(JBGS(2021)109;JBGS[2021]029);国家自然科学基金项目(32102538);广东省基础与应用基础研究基金项目(2022A1515012014);扬州市现代农业项目(YZ2023054)。

摘　　要：为探究gga-miR-17-5p在鸡脂肪沉积和肌肉发育中的作用,本研究以矮小品系S3(DW)和隐性白羽鸡(RR)为试验素材,分析gga-miR-17-5p在不同时期腿肌、肝脏、腹脂组织及细胞中的表达差异,并通过KEGG及蛋白互作分析筛选关键靶基因。结果表明:1)gga-miR-17-5p在腹脂、肝脏和腿肌组织中均可表达。在腹脂中,gga-miR-17-5p在0周表达有品种差异(P<0.05),且2个品种在0周时显著高于其他周龄(P<0.05);在肝脏中,gga-miR-17-5p在16周有品种差异(P<0.05),S3系在16周显著高于其他周龄(P<0.05),隐性白羽肉鸡在16周显著高于0周和8周(P<0.05);在腿肌中,gga-miR-17-5p在0周有品种差异(P<0.05),S3系在0周和2周显著于其它周龄(P<0.05),隐性白羽肉鸡在0周显著高于其他周龄(P<0.05)。2)在脂肪细胞中,gga-miR-17-5p在肌内脂肪细胞诱导分化6 d后的表达显著高于增殖期和分化1 d(P<0.05);腹脂细胞分化1 d的表达最低,显著低于增殖期、分化4 d和分化6 d(P<0.05)。3)在成肌细胞中,gga-miR-17-5p的表达量随分化时间的延长而升高,且分化6 d的表达显著高于增殖期(P<0.05)。4)KEGG及蛋白互作分析表明,PTEN、MAPK3、PIK3R1、BECN1和PLCB4是gga-miR-17-5p重要的靶基因。RT-qPCR结果表明,PTEN和PIK3R1在成肌细胞增殖期的表达显著高于分化期(P<0.05)。综上,gga-miR-17-5p的表达存在显著的品种和组织差异性,miR-17-5p很可能通过PTEN、MAPK3、PIK3R1、BECN1和PLCB4来影响肌肉发育和脂肪沉积,其中PTEN和PIK3R1尤为关键。本研究为深入研究gga-miR-17-5p的功能和机制提供了坚实的理论基础和数据支撑。To investigate the role of gga-miR-17-5p in chicken fat deposition and muscle development,the study used dwarf strain S3(DW)and recessive white feathered chickens(RR)as test materials to analyze the expression differences of gga-miR-17-5p in bone and adipose tissues and cells at different periods of time,and to screen the key target genes by KEGG and protein interactions analysis.The results showed that:1)gga-miR-17-5p was expressed in abdominal fat,liver and leg muscle tissues.In abdominal fat,there was a variety difference in the expression of gga-miR-17-5p at 0 week(P<0.05),and the two varieties were significantly higher in two breeds at week 0 than other weekly ages(P<0.05).In liver,gga-miR-17-5p was expressed with breed differences at week 16(P<0.05),and was significantly higher in S3 line at week 16 than other weekly ages(P<0.05),and was significantly higher in cryptic white-feathered broilers at 16 weeks was significantly higher than at 0 and 8 weeks(P<0.05).In the leg muscle,there was a breed difference in gga-miR-17-5p at 0 weeks(P<0.05),and the S3 line was significantly higher than other weeks at 0 and 2 weeks(P<0.05),The recessive white feathered chickens were significantly higher than other weeks of age at 0 weeks(P<0.05).2)In adipocytes,the expression of gga-miR-17-5p in intramuscular adipocytes after 6 days of induction of differentiation was significantly higher than that in the proliferation phase and 1 day of differentiation.(P<0.05)The lowest expression was found in abdominal adipocytes at 1 d of differentiation,which was significantly lower than that in the proliferative stage,at 4 d of differentiation,and at 6 d of differentiation(P<0.05).3)In myoblasts,the expression of gga-miR-17-5p was elevated with the prolongation of the time of differentiation,and it was significantly higher in 6 d of differentiation than that in the proliferative stage(P<0.05).4)KEGG and protein interactions analysis showed that PTEN,MAPK3,PIK3R1,BECN1 and PLCB4 were important target genes of gga-miR-17-5p.The RT-qPCR r

关 键 词：鸡 gga-miR-17-5p 脂肪沉积 肌肉发育 

分 类 号：S831[农业科学—畜牧学]