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半滑舌鳎雌、雄鱼myostatin基因差异表达的初步研究
张梦倩,崔忠凯,陈张帆,徐文腾,陈松林
1.中国水产科学研究院黄海水产研究所 农业农村部海洋渔业可持续发展重点实验室 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 山东省海洋渔业生物技术与遗传育种重点实验室 山东 青岛 266071;2.上海海洋大学水产科学国家级实验教学示范中心 上海 201306;3.中国水产科学研究院黄海水产研究所 农业农村部海洋渔业可持续发展重点实验室 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 山东省海洋渔业生物技术与遗传育种重点实验室 山东 青岛 266071
摘要:
半滑舌鳎(Cynoglossus semilaevis)生长具有明显的性别二态性,性成熟雌鱼体重显著高于雄鱼。鱼体肌肉占体重的40%~60%,肌肉生长是生长性状的核心,而myostatin (mstn)基因是肌肉生长发育的负调控因子,对成肌细胞的增殖具有抑制作用。本研究旨在探究肌肉生长发育过程中,mstn基因在半滑舌鳎雌、雄鱼的时空表达模式及其与雌、雄鱼生长差异的关系。结果显示,在所检测的1龄鱼的9个组织中,mstn在肌肉中的表达量最高。不同发育时期的肌肉组织定量结果显示,1龄和2龄雄鱼中的表达量显著高于雌鱼(P<0.05),而在选取的其他时期,雌、雄鱼之间的表达量不存在显著差异。原位杂交结果显示,mstn基因的表达位置在肌纤维边缘,即成肌细胞增殖分化的位置。本文系统研究了肌肉生长发育的负调控因子mstn在半滑舌鳎雌、雄鱼中的时空表达模式,为深入研究半滑舌鳎雌、雄鱼生长差异的原因及其机制提供了基础。
关键词:  半滑舌鳎  myostatin  肌肉生长发育  qRCR  原位杂交
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Gender differential expression of myostatin gene in Chinese tongue sole (Cynoglossus semilaevis)
ZHANG Mengqian1,2,3,4,5, CUI Zhongkai1,2,3,4, CHEN Zhangfan1,2,3,4, XU Wenteng1,2,3,4, CHEN Songlin1,2,3,4
1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;2.Key Laboratory for Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs;3.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao Pilot National Laboratory for Marine Science and Technology;4.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao, Shandong 266071, China;5.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China
Abstract:
Chinese tongue sole (Cynoglossus semilaevis) growth exhibits clear sex dimorphism, and the weight of mature female fish is significantly greater than that of male fish. In fish, muscle accounts for 40%-60% of body weight, and muscle growth is the core growth trait. Myostatin (mstn), also known as GDF-8, is a member of the transcriptional growth factor (TGF-β) superfamily. mstn is a negative regulator of muscle growth and development and has an inhibitory effect on myoblast proliferation. mstn gene deletion can lead to an increase in skeletal muscle mass. In recent years, research on the mstn gene has primarily focused on the aspects of gene cloning, expression pattern analysis, and RNA interference. Minimal studies have reported the differential expression of the mstn gene between male and female fish with sex dimorphism. The aim of this study was to explore the spatiotemporal expression pattern of the mstn gene in different sexes of Chinese tongue sole during muscle growth and development. The results showed that mstn expression was detected in the liver, spleen, intestine, kidney, gonad, gill, heart, brain, and muscle of 1-year-old fish, and mstn expression was the highest in muscle. The quantitative results of muscle tissue at different developmental stages showed that the expression levels in one- and two-year-old males were significantly higher than those in females (P<0.05), however, there was no significant difference in expression levels between males and females in other selected stages. Tissue section staining showed that the muscle fiber cross sections presented an irregular polygonal shape. The results of in situ hybridization showed that mstn gene expression was found at the edge of the muscle fiber, where myoblasts proliferate and differentiate. In this study, the spatial and temporal expression patterns of mstn were systematically studied in different sexes of Chinese tongue sole, which provides a basis for further studies on the growth differences between male and female Chinese tongue sole.
Key words:  Cynoglossus semilaevis  myostatin  Muscle growth and development  qPCR  in situ hybridization