1.华中农业大学水产学院/农业农村部淡水生物繁育重点实验室/农业动物遗传育种与繁殖教育部重点实验室，武汉 430070;2.长江经济带大宗水生生物产业绿色发展教育部工程研究中心/ 湖北省名优鱼育种与健康养殖工程技术研究中心，武汉 430070
邹雪，E-mail： zousnow@webmail.hzau.edu.cn
刘露莎，E-mail： liuls@mail.hzau.edu.cn
Q959.46+8
国家自然科学基金联合基金项目（U21A20263）；中央高校基本科研业务费专项（2662022SCQD001）；国家大宗淡水鱼类产业技术体系建设项目（CARS-45-01）
- ZOU Xue 1
- LIU Qi 1
- ZHOU Jiajia 1
- WU Tianyu 1
- LIU Lusha 1
- GAO Zexia 1,2
GAO Zexia
Key Lab of Freshwater Animal Breeding， Ministry of Agriculture and Rural Affairs/ Key Lab of Agricultural Animal Genetics， Breeding and Reproduction of Ministry of Education/ College of Fisheries， Huazhong Agricultural University， Wuhan 430070， China;Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt， Ministry of Education/ Hubei Province Famous Fish Breeding and Healthy Aquaculture Engineering Technology Research Center， Wuhan 430070， China
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1.Key Lab of Freshwater Animal Breeding， Ministry of Agriculture and Rural Affairs/ Key Lab of Agricultural Animal Genetics， Breeding and Reproduction of Ministry of Education/ College of Fisheries， Huazhong Agricultural University， Wuhan 430070， China;2.Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt， Ministry of Education/ Hubei Province Famous Fish Breeding and Healthy Aquaculture Engineering Technology Research Center， Wuhan 430070， China
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为探究myomaker基因在团头鲂（Megalobrama amblycephala）肌纤维发育中的调控作用，通过石蜡切片和HE染色方法分析不同日龄团头鲂肌纤维发育特征，采用RT-PCR及qRT-PCR技术鉴定myomaker基因的cDNA序列，并探讨该基因在团头鲂不同发育阶段及不同组织部位的时空表达模式。石蜡切片结果显示团头鲂肌纤维直径在20~60 d持续递增，其中在40、50和60 d均显著递增（P<0.05），且在40~50 d时其肌纤维直径递增最快。基因结构分析发现团头鲂myomaker基因全长4 693 bp，包含5个外显子和4个内含子，开放阅读框为663 bp，共编码220个氨基酸，其编码蛋白为7次跨膜蛋白。氨基酸相似度比对显示myomaker基因保守性较高；系统进化树分析显示，团头鲂myomaker基因与鲤等鲤科鱼类的myomaker基因聚为一支，具有最近的亲缘关系。荧光定量结果显示，myomaker在1月龄团头鲂肌肉组织中表达量极其显著高于其他组织（P<0.000 1），同时myomaker在团头鲂出膜后15~30 d表达量逐渐上升，在30 d表达量最高（P<0.05）。基于形态学和基因表达特征，推测myomaker基因在团头鲂30 d后肌纤维的融合中发挥重要的调控作用，从而促进肌纤维的肥大。
To investigate the role of the myomaker gene in muscle hypertrophy in Megalobrama amblycephala， the cDNA sequence was identified using RT-PCR. The developmental characteristics of muscle fibers at various stages in M. amblycephala were analyzed through paraffin sections and HE staining. Additionally， the expression patterns of the myomaker gene at different developmental stages and in different tissues of M. amblycephala were examined using real-time PCR. The results showed that the length of the myomaker gene in M. amblycephala is 4 693 bp， containing 5 exons and 4 introns. The open reading frame of myomaker is 663 bp and encodes a protein of 220 amino acids with 7 transmembrane domains. An analysis of the amino acid sequences revealed that the myomaker protein sequences were highly conserved throughout evolution， and phylogenetic tree analysis demonstrated that the myomaker of M. amblycephala clustered with that of cyprinid fish， indicating a close evolutionary relationship. The results of paraffin sections showed that the diameter of muscle fibers was significantly increased from 30 d to 60 d （P<0.05）， with the most rapid growth occurring between 40 d and 50 d. The real-time PCR results of muscle tissue at different developmental stages showed a gradual increase in expression from 15 to 30 days post hatching， peaking at 30 d （P<0.05）. Combined with morphological data， these findings suggest that the myomaker gene plays an important role in muscle fiber fusion after 30 d， thereby promoting muscle fiber hypertrophy. Furthermore， the real-time PCR results indicated that the expression level of myomaker in the muscle tissue of one-month-old M. amblycephala was significantly higher than that in other tissues （P<0.000 1）. These results demonstrate that the myomaker gene plays a significant regulatory role in the hypertrophy process of muscle fibers in M. amblycephala.
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邹雪,刘起,周佳佳,吴天宇,刘露莎,高泽霞.团头鲂myomaker基因的鉴定及其在肌纤维发育中的功能[J].华中农业大学学报,2025,44(1):201-210
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- 收稿日期:2023-10-09
- 在线发布日期: 2025-03-03
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