广西麻鸡m6A甲基转移酶基因表达与肌纤维类型及成肌分化的关系  

Relationship Between Expression Levels of Guangxi Partridge Chicken m6A Methyltransferase Genes,Myofiber Types and Myogenic Differentiation

作　　者：束婧婷[1] 单艳菊[1] 姬改革[1] 章明[1] 屠云洁[1] 刘一帆[1] 巨晓军 盛中伟[1] 唐燕飞 李华[3] 邹剑敏[1] SHU JingTing;SHAN YanJu;JI GaiGe;ZHANG Ming;TU YunJie;LIU YiFan;JU XiaoJun;SHENG ZhongWei;TANG YanFei;LI Hua;ZOU JianMin(Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province,Jiangsu Institute of Poultry Science,Yangzhou 225125,Jiangsu;Guangxi Fufeng Farm Group Co.,Ltd.Nanning 530024;Foshan University,Foshan 528225,Guangdong)

机构地区：[1]江苏省家禽科学研究所江苏省家禽遗传育种重点实验室,江苏扬州225125 [2]广西富凤农牧集团有限公司,南宁530024 [3]佛山科学技术学院,广东佛山528225

出　　处：《中国农业科学》2022年第3期589-601,共13页Scientia Agricultura Sinica

基　　金：江苏省农业自主创新资金(CX(20)3003);现代农业产业技术体系专项资金(CARS-41);江苏省种业创新揭榜挂帅项目(JBGS[2021]107);江苏省农业重大新品种创制项目(PZCZ201728);江苏现代农业(肉鸡)产业技术体系集成创新中心(JATS[2021]396);江苏省自然科学基金(BK20211121,BK20210955)。

摘　　要：【目的】肌纤维类型及其转化的调节是改善肉质的重要途径,最新研究表明,m6A RNA甲基化修饰在肌肉的分化中发挥着重要作用。以优良的地方品种--广西麻鸡作为研究对象,研究其性成熟日龄不同部位肌纤维类型的组成差异,探究m6A甲基化转移酶相关基因甲基化转移酶样蛋白3/14(methyltransferase like 3/14,METTL3/14)、Wilm肿瘤1-相关蛋白(Wilms’tumor 1-associating protein,WTAP)和病毒样m6A甲基转化酶(vir like m6A methyltransferase associated,VIRMA,也称KIAA1429)在不同部位肌肉组织和成肌细胞分化前后的表达差异,为阐明肌纤维类型组成及转换的调控机理提供参考。【方法】采用ATPase染色法比较广西麻鸡胸部、腿部和背部肌肉群7个部位肌纤维类型、直径和密度等肌纤维性状差异,采用比色法检测成肌细胞分化前后不同时间点内源性m6A甲基化水平,采用实时荧光定量PCR方法检测上述4个基因在广西麻鸡7个部位肌肉以及成肌细胞分化前后的表达差异,并将其与肌纤维性状和m6A甲基化水平进行相关性分析。【结果】广西麻鸡性成熟日龄胸部肌肉群的胸大肌和胸小肌全部由白肌纤维组成,腿部肌肉群的耻坐骨肌内侧头、腓肠肌内侧头和缝匠肌均以红肌纤维为主,而髂胫外侧肌以白肌纤维为主,背部肌肉群的背阔肌则以红肌纤维为主;总体上,红肌纤维比例多的肌肉肌纤维密度显著高于白肌纤维比例多的肌肉。METTL3、METTL14、WTAP和KIAA1429基因表达存在显著的组织和时间特异性,总体上,在红肌纤维为主的肌肉中的表达量高于白肌纤维为主的肌肉,在分化期成肌细胞中的表达量要显著高于增殖期成肌细胞。肌肉中METTL3、METTL14和WTAP具有相似的表达模式,均在背阔肌中表达量最高,显著高于其他肌肉组织;在耻坐骨肌内侧头和腓肠肌内侧头的表达量次之,显著高于缝匠肌、胸大肌、胸小肌和髂胫外侧肌。K【Objective】It is widely accepted that the regulation of myofiber type composition and transition is an important source of variation to improve meat quality.Recently,several studies revealed that N6-methyladenosine(m6A)RNA methylation played a important biological role in the regulation of muscle differentiation.In the present study,the myofiber types of different location of skeletal muscles were identified in the native chicken breed-Guangxi partridge chicken,and the expression levels of the m6A methyltransferase genes(METTL3,METTL14,WTAP and KIAA1429)were first studied in different location of skeletal muscle and myogenic differentiation,which would provide a reference for clarifying the regulation mechanism of chicken myofiber type composition and transition.【Method】Myosin ATPase staining was used to identify the myofiber types and to measure the myofiber size and density of seven location of Guangxi partridge chicken muscles,including the Pectoralis major(XD),pectoralis minor(XX),Sartorius(FJ),Medial head of sciatic pubis(CN),medial head of gastrocnemius(FN),Lateral iliotibial muscle(QW)and Latissimus dorsi(BK).Colorimetric method was used to detect the endogenous m6A methylation levels at different time points before and after myogenic differentiation.Expression of METTL3,METTL14,WTAP and KIAA1429 was quantified by RT-PCR in these seven muscles,as well as at the myoblasts separated on 0 h,19 h,and cultured in differentiation medium on D0,D2,and D5,and the correlations between the gene expression levels and myofiber characteristics as well as m6A methylation levels were also analyzed by SPSS20.0 software.【Result】The XD and XX muscles of the breast muscle group were all composed of white myofibers,CN,FN and FJ muscles of the leg muscle group were mainly composed of red myofibers,and QW muscles were mainly composed of white myofibers,while the BK muscles of the back muscle group were mainly composed of red myofibers.The average fiber densities of muscles mainly composed of red myofibers were signif

关 键 词：广西麻鸡 基因表达 m6A甲基化转移酶基因 肌纤维类型 成肌分化 

分 类 号：S831.2[农业科学—畜牧学]