The production of fluorescent transgenic trout to study in vitro myogenic cell differentiation

BMC Biotechnology

Research Article

Cécile Rallière¹  Jean-Charles Gabillard¹  Nathalie Sabin¹  Pierre-Yves Rescan¹ 

[1] National Institute for Agricultural Research, Joint Research Unit for Fish Physiology, Biodiversity and the Environment, INRA Scribe, IFR140, Campus de Beaulieu, 35042, Rennes, France;

关键词: Satellite Cell;    Muscle Growth;    Myogenic Cell;    Muscle Cell Differentiation;    Multinucleated Myotubes;   

DOI  :  10.1186/1472-6750-10-39

 received in 2009-09-22, accepted in 2010-05-17,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundFish skeletal muscle growth involves the activation of a resident myogenic stem cell population, referred to as satellite cells, that can fuse with pre-existing muscle fibers or among themselves to generate a new fiber. In order to monitor the regulation of myogenic cell differentiation and fusion by various extrinsic factors, we generated transgenic trout (Oncorhynchus mykiss) carrying a construct containing the green fluorescent protein reporter gene driven by a fast myosin light chain 2 (MlC2f) promoter, and cultivated genetically modified myogenic cells derived from these fish.ResultsIn transgenic trout, green fluorescence appeared in fast muscle fibers as early as the somitogenesis stage and persisted throughout life. Using an in vitro myogenesis system we observed that satellite cells isolated from the myotomal muscle of transgenic trout expressed GFP about 5 days post-plating as they started to fuse. GFP fluorescence persisted subsequently in myosatellite cell-derived myotubes. Using this in vitro myogenesis system, we showed that the rate of muscle cell differentiation was strongly dependent on temperature, one of the most important environmental factors in the muscle growth of poikilotherms.ConclusionsWe produced MLC2f-gfp transgenic trout that exhibited fluorescence in their fast muscle fibers. The culture of muscle cells extracted from these trout enabled the real-time monitoring of myogenic differentiation. This in vitro myogenesis system could have numerous applications in fish physiology to evaluate the myogenic activity of circulating growth factors, to test interfering RNA and to assess the myogenic potential of fish mesenchymal stem cells. In ecotoxicology, this system could be useful to assess the impact of environmental factors and marine pollutants on fish muscle growth.

【 授权许可】

Unknown   
© Gabillard et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.