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 results showed that the expression of PTEN and PIK3R1 was significantly higher in the proliferative stage of myoblasts than that in the differentiation stage(P<0.05).Taken together,the expression of gga-miR-17-5p exhibits significant species and tissue variability and miR-17-5p is likely to affect muscle development and fat deposition through PTEN,MAPK3,PIK3R1,BECN1 and PLCB4,of which PTEN and PIK3R1 are particularly critical.This study provides a solid theoretical foundation for the in-depth study of gga-miR-17-5p function and mechanism.
作者
杨苗苗
向海
李瑞瑞
赵振华
王钱保
黄正洋
李春苗
梁忠
吴兆林
黄华云
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)
出处
《中国农业大学学报》
CAS
CSCD
北大核心
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)。
作者简介
第一作者:杨苗苗,佛山科学技术学院在读硕士研究生,现于江苏省家禽研究所联合培养,主要从事动物遗传育种工作。2022-2023年两年获得二等、三等学业奖学金(ORCID:0009-0001-9275-3207),E-mail:1079278298@qq.com;通讯作者:黄华云,博士,研究员,江苏省家禽科学研究所肉鸡育种与生产科室副主任。主要从事家禽遗传育种与生产技术研究。中国畜牧兽医学会家禽学分会理事、江苏省第五期、第六期“333高层次人才培养工程”第三层次培养对象。主持农业生物育种重大项目子课题、江苏省自然科学基金面上基金、江苏省农业自主创新、扬州市现代农业等项目7项,荣获江苏省科学技术奖二等奖、全国农牧渔业丰收奖二等奖、江苏省科学技术奖三等奖,在JIA等期刊发表SCI论文7篇,中文核心期刊十余篇,参与授权发明专利20余项。(ORCID:0000-0002-1192-3674),E-mail:huanghuayun520@163.com。
