摘要
旨在了解m6A甲基转移酶样蛋白16(methyltransferase-like protein 16,METTL16)在鸡不同类型肌肉组织中的表达情况及其在鸡骨骼肌功能中的调控作用。本研究利用qRT-PCR技术检测METTL16基因在120日龄广西麻鸡母鸡不同类型肌肉组织中的表达,同时利用siRNA干扰鸡原代成肌细胞中METTL16基因的表达,分析其对成肌细胞增殖、分化和肌纤维形成的影响。结果显示,METTL16基因在鸡的不同类型肌肉组织(胸大肌、胸小肌、缝匠肌、耻坐骨肌内侧肌、耻坐骨肌外侧肌、髂胫外侧肌、腓肠肌内侧肌和背阔肌)中广泛表达,其中,在白肌纤维为主的胸大肌、胸小肌和髂胫外侧肌中表达量相对更高;细胞增殖检测结果显示,干扰METTL16基因表达后,鸡成肌细胞的增殖活力受到了抑制;在成肌细胞分化后干扰METTL16基因表达,鸡成肌细胞RNA m6A甲基化水平呈下降趋势,细胞分化关键基因MyoD表达显著下降(P<0.05),fast-MyHC基因表达显著上升(P<0.001),slow-MyHC基因表达呈下降趋势。综上,METTL16基因在不同类型肌肉中的表达与其肌纤维类型组成相关,METTL16基因可能在鸡成肌细胞增殖和分化中发挥重要作用,并抑制快肌纤维的形成,提示METTL16可能在鸡肌肉功能调控中发挥重要作用。
The purpose of this study was to explore the expression of m6A methyltransferase-like protein 16(METTL16) in different types of chicken muscle tissue and its regulatory role in chicken skeletal muscle function. In the present study, the tissue expression profile of METTL16 in different muscles of 120-day-old Guangxi partridge hens and the expression of genes related to proliferation, differentiation and muscle fiber formation after siRNA interference by qRT-PCR were analyzed. The results showed that METTL16 gene was widely expressed in different muscle tissues of chickens. White muscles, including major pectoralis, minor pectoralis and lateral iliotibial muscles, have higher METTL16 gene expression. Cell proliferation assay showed that the proliferation activity of chicken myoblasts was inhibited after METTL16 interference. Interfering with METTL16 gene during myoblast differentiation could reduced the methylation level of m6A and slow-MyHC expression in chicken myoblasts, significantly reduced MyoD expression(P<0.05), and significantly elevated fast-MyHC expression(P<0.001). In conclusion, the expression of METTL16 gene in different muscles is related to the composition of muscle fiber types. METTL16 gene may play a significant role in chicken myoblast proliferation and differentiation, and inhibit the formation of fast-twitch muscle fibers. These results indicated that METTL16 might play a crucial role in chicken muscle function regulation.
作者
庞立川
单艳菊
刘一帆
章明
甘达峰
屠云洁
姬改革
巨晓军
束婧婷
邹剑敏
PANG Lichuan;SHAN Yanju;LIU Yifan;ZHANG Ming;GAN Dafeng;TU Yunjie;JI Gaige;JU Xiaojun;SHU Jingting;ZOU Jianmin(Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province,Jiangsu Institution of Poultry Science,Yangzhou 225125,China;Chongren Animal Breeding Farm,Fuzhou 344200,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2023年第2期545-553,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
现代农业产业技术体系建设专项资金(CARS-41)
江苏省自然科学基金(BK20211121)
江苏省种业振兴“揭榜挂帅”(JBGS[2021]107)
江苏省农业重大新品种创制项目(PZCZ201728)。
作者简介
庞立川(1994-),男,山东潍坊人,硕士生,主要从事鸡遗传育种研究,E-mail:945907249@qq.com;通信作者:邹剑敏,主要从事家禽遗传育种研究,E-mail:jqszjm@163.com;通信作者:束婧婷,主要从事家禽遗传育种研究,E-mail:shujingting@163.com。
