Gaffney Christopher J, Bass Joseph J, Barratt Thomas F, Szewczyk Nathaniel J
MRC/ARUK Centre for Musculoskeletal Ageing Research, University of Nottingham.
MRC/ARUK Centre for Musculoskeletal Ageing Research, University of Nottingham;
J Vis Exp. 2014 Nov 13(93):e52043. doi: 10.3791/52043.
Muscle is a dynamic tissue that responds to changes in nutrition, exercise, and disease state. The loss of muscle mass and function with disease and age are significant public health burdens. We currently understand little about the genetic regulation of muscle health with disease or age. The nematode C. elegans is an established model for understanding the genomic regulation of biological processes of interest. This worm's body wall muscles display a large degree of homology with the muscles of higher metazoan species. Since C. elegans is a transparent organism, the localization of GFP to mitochondria and sarcomeres allows visualization of these structures in vivo. Similarly, feeding animals cationic dyes, which accumulate based on the existence of a mitochondrial membrane potential, allows the assessment of mitochondrial function in vivo. These methods, as well as assessment of muscle protein homeostasis, are combined with assessment of whole animal muscle function, in the form of movement assays, to allow correlation of sub-cellular defects with functional measures of muscle performance. Thus, C. elegans provides a powerful platform with which to assess the impact of mutations, gene knockdown, and/or chemical compounds upon muscle structure and function. Lastly, as GFP, cationic dyes, and movement assays are assessed non-invasively, prospective studies of muscle structure and function can be conducted across the whole life course and this at present cannot be easily investigated in vivo in any other organism.
肌肉是一种动态组织，会对营养、运动和疾病状态的变化做出反应。随着疾病和年龄增长，肌肉质量和功能的丧失是重大的公共卫生负担。目前，我们对疾病或年龄相关的肌肉健康的基因调控了解甚少。秀丽隐杆线虫是一种成熟的模型，用于理解感兴趣的生物过程的基因组调控。这种线虫的体壁肌肉与高等后生动物物种的肌肉具有高度同源性。由于秀丽隐杆线虫是一种透明生物，绿色荧光蛋白（GFP）定位于线粒体和肌节，使得这些结构在体内可视化。同样，给动物喂食阳离子染料，这些染料会根据线粒体膜电位的存在而积累，从而可以在体内评估线粒体功能。这些方法，以及对肌肉蛋白质稳态的评估，与以运动测定形式进行的全动物肌肉功能评估相结合，以便将亚细胞缺陷与肌肉性能的功能指标相关联。因此，秀丽隐杆线虫提供了一个强大的平台，用于评估突变、基因敲低和/或化合物对肌肉结构和功能的影响。最后，由于对绿色荧光蛋白、阳离子染料和运动测定的评估是非侵入性的，因此可以在整个生命过程中对肌肉结构和功能进行前瞻性研究，而目前在任何其他生物体中都不容易在体内进行此类研究。
