title: The isolated muscle fibre as a model of disuse atrophy: characterization using PhAct, a method to quantify f-actin.
abstract: Research into muscle atrophy and hypertrophy is hampered by limitations of the available experimental models. Interpretation of in vivo experiments is confounded by the complexity of the environment while in vitro models are subject to the marked disparities between cultured myotubes and the mature myofibres of living tissues. Here we develop a method (PhAct) based on ex vivo maintenance of the isolated myofibre as a model of disuse atrophy, using standard microscopy equipment and widely available analysis software, to measure f-actin content per myofibre and per nucleus over two weeks of ex vivo maintenance. We characterize the 35% per week atrophy of the isolated myofibre in terms of early changes in gene expression and investigate the effects on loss of muscle mass of modulatory agents, including Myostatin and Follistatin. By tracing the incorporation of a nucleotide analogue we show that the observed atrophy is not associated with loss or replacement of myonuclei. Such a completely controlled investigation can be conducted with the myofibres of a single muscle. With this novel method we can distinguish those features and mechanisms of atrophy and hypertrophy that are intrinsic to the muscle fibre from those that include activities of other tissues and systemic agents.
authors.affiliation: Center for Genetic Medicine, Children's Research Institute, Children's National Medical Center, Washington, DC, USA. bduddy@cnmcresearch.org
authors.forename: William J
authors.initials: WJ
authors.lastname: Duddy
authors.forename: Tatiana
authors.initials: T
authors.lastname: Cohen
authors.forename: Stephanie
authors.initials: S
authors.lastname: Duguez
authors.forename: Terence A
authors.initials: TA
authors.lastname: Partridge
mesh_terms: D000199:Actins / Q000032:analysis* / Q000235:genetics; D000818:Animals; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000378:metabolism / Q000473:pathology*; D009133:Muscular Atrophy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012984:Software
chemical_list: D000199:Actins; D012333:RNA, Messenger