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作 者:于倩文 黄可佳 张全启[1,2,3] 贺艳 Yu Qianwen;Huang Kejia;Zhang Quanqi;He Yan(Key Laboratory of Marine Genetics and Breeding(Ocean University of China),Ministry of Education,Qingdao 266003,China;Laboratory of Tropical Marine Germplasm Resources and Breeding Engineering,Sanya Oceanographic Institution,Ocean University of China,Sanya 572000,China;Pilot National Laboratory for Marine Science and Technology(Qingdao),Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao 266237,China)
机构地区:[1]中国海洋大学海洋生物遗传学与育种教育部重点实验室,山东青岛266003 [2]中国海洋大学三亚海洋研究院海南省热带水产种质重点实验室,海南三亚572000 [3]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,山东青岛266237
出 处:《中国海洋大学学报(自然科学版)》2024年第1期67-78,共12页Periodical of Ocean University of China
基 金:国家重点基础研究发展计划项目(2018YFD0900101)资助。
摘 要:为探究Myomaker在硬骨鱼肌肉生长过程中调控作用,本研究以肌肉具有无限生长能力的许氏平鲉(Sebastes schlegelii)为研究对象,证明Myomaker在其肌肉生长过程中发挥重要作用。许氏平鲉myomaker基因全长5386 bp,编码序列长870 bp,编码289个氨基酸。胚胎整体原位杂交实验结果表明,myomaker的表达始于体节期,信号主要集中在体节位置,而在孵化前期和仔鱼期信号主要集中在头部和躯干前部。用myomaker过表达质粒投喂仔鱼30 d后,myomaker表达量显著升高,且过表达组小面积肌纤维(500~1000μm^(2))数目显著少于对照组,而大面积肌纤维(>1000μm^(2))数目显著多于对照组。之后停止投喂myomaker过表达质粒,继续培养90 d,过表达组大面积肌纤维(>5000μm^(2))占比仍大于对照组,这说明Myomaker在促进许氏平鲉肌肉肥大生长方面发挥了重要作用。此外,体外细胞实验证明许氏平鲉Myomaker可以促进小鼠C2C12成肌细胞发生融合,这说明Myomaker促进许氏平鲉肌纤维面积增大可能是通过调控成肌细胞融合实现的。本研究结果丰富了Myomaker调控非模式动物肌肉生长发育的资料,为进一步加深对大体型硬骨鱼类肌肉无限生长调控分子机理的理解奠定了基础。To explore the regulatory role of myomaker during muscle growth in teleosts,Sebastes schlegelii,an important marine economic fish species in northern China was used in this study,which displays indeterminate muscle growth,and demonstrated that myomaker gene plays an important role in its muscle growth.The full length of the myomaker gene of S.schlegelii is 5386 bp,with coding sequence of 870 bp,encoding a protein of 289 amino acids.Whole-mount in situ hybridization of S.schlegelii embryos revealed that myomaker expression was initiated at the somite stage and showed intense expression exclusively in the developing somites.However,in the pre-hatching and larval stages the signal was mainly concentrated in the head and anterior part of trunk.After overexpression of myomaker by feeding larval S.schlegelii with plasmid for 30 days,the expression of myomaker was significantly elevated.myomaker could promote hypertrophy growth of muscle based on the evidence that the overexpression group contained significantly fewer number of small-scale fibers(500~1000μm^(2))and more of large-scale fibers(>1000μm^(2))than those in the control group.The ratio of large-scale fibers(>5000μm^(2))in the overexpression group is still larger than that in the control group after 90 days culture post the termination of overexpression treatment.In addition,S.schlegelii myomaker could drive the fusion of C2C12 myoblasts.It is speculated that myomaker drives hypertrophy growth of muscle in S.schlegelii by promoting myoblast fusion.This study highlights the important role of myomaker in regulating muscle growth in nonmodel species and provides a foundation to our understanding of the mechanisms involved in the indeterminate growth in large scale teleost fish.
关 键 词:许氏平鲉 myomaker基因 肌纤维肥大生长 成肌细胞融合
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