摘要
为解析肌肉生长抑制素(Myostatin,MSTN)基因在黄条鰤(Seriola aureovittata)生长发育过程中作用机制。本研究采用cDNA末端快速扩增法,克隆了黄条鰤MSTN基因的全长cDNA序列,采用荧光定量PCR(RT-qPCR)技术对MSTN mRNA在黄条鰤不同组织、胚胎发育过程以及仔稚幼鱼发育过程中的差异表达特征进行了研究。序列分析结果显示,MSTN基因全长cDNA为1828 bp,开放阅读框为1131 bp,编码了376个氨基酸。RT-qPCR分析表明,在检测的脑、垂体、肝脏、肌肉、脾脏、肾脏、鳃、心脏、胃、肠、头肾组织中MSTN mRNA均有表达,其在肌肉中表达量最高(P<0.05)。MSTN mRNA在胚体下包2/3时期至孵化期表达量显著升高,且在孵化期达到峰值(P<0.05)。MSTN mRNA在孵化后3天前表达量显著升高(P<0.05),在第20天降低至较低水平,25天后表达量再次显著升高,在第30天达到峰值(P<0.05)。研究结果首次揭示了MSTN基因在黄条鰤的胚胎及早期生长发育过程中发挥着重要作用,本研究为开展黄条鰤繁养殖、育种提供了理论参考。
Myostatin(MSTN)is a member of the transforming growth factor-β(TGF-β)superfamily that functions as a negative regulator in the muscle growth and development of animals.To understand the regulatory roles of MSTN in the growth and development of S.aureovittata,the full-length cDNA sequences of MSTN was isolated with the rapid amplification of cDNA ends(RACE)method.The real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression of MSTN in different tissues,at embryonic development stages,and at larval and juvenile stages in this study.The full length cDNA was1828 bp in length and the open reading frame was 1131 bp(GenBank accession number,MH144053)which encoded 376 amino acids.The putative MSTN of S.aureovittata had the essential characteristic of vertebrate MSTN.qRT-PCR analysis demonstrated that the MSTN differentially expressed in brain,pituitary,liver,muscle,spleen,kidney,gill,heart,stomach,intestine and head-kidney of adult S.aureovittata with the highest in muscle(P<0.05).The MSTN was confirmed to express at every stage of embryogenesis of S.aureovittata.During the embryonic development,the expression of MSTN increased significantly in the embryo encircling 2/3 of yolk sac,and peaked in the hatching larva(P<0.05).At the larval and juvenile stages,the expression of MSTN was significantly up-regulated at 3 days post-hatching(dph),then decreased to a lower level at 20 dph(P<0.05).The expression of MSTN significantly increased at 25 dph and peaked at 30 dph(P<0.05).Taken toget-her,these results provided the first evidence that MSTN plays important regulatory roles at embryo and early developmental stage and also a theoretical basis for reproduction,culture and breeding of S.aureovittata.
作者
史宝
曹亚男
柳学周
孙冉冉
徐永江
王滨
姜燕
SHI Bao;CAO Ya-Nan;LIU Xue-Zhou;SUN Ran-Ran;XU Yong-Jiang;WANG Bin;JIANG Yan(Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology(Qingdao), Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Yantai Marine Economic Research Institute, Yantai 264000, China;College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China)
出处
《中国海洋大学学报(自然科学版)》
CAS
CSCD
北大核心
2021年第4期35-43,共9页
Periodical of Ocean University of China
基金
青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室开放课题(2017-3A01)
国家重点研究发展计划项目(2018YFD0901204,2019YFD0900503)
国家自然科学基金项目(31772829)
中国水产科学研究院院级基本科研业务费-农业部海洋渔业可持续发展重点实验室开放课题(2019HY-XKQ01)
国家海水鱼产业技术体系项目(CARS-47)资助。
关键词
黄条鰤
MSTN基因
胚胎发育
仔稚幼鱼发育
基因克隆
表达特征
Seriola aureovittata
MSTN gene
embryonic development
larval and juvenile development
cDNA isolation
expression analysis