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Isolated Murine Myofibres Undergo Atrophy Ex Vivo Via Diminution of the Myonuclear Domain
Author(s) -
Duddy William John,
Cohen Tatiana,
Duguez Stephanie,
Partridge Terence
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1051.20
Subject(s) - ex vivo , myogenesis , myostatin , in vivo , atrophy , microbiology and biotechnology , muscle atrophy , biology , skeletal muscle , myocyte , pathology , anatomy , genetics , medicine
The interpretations of in vivo experiments on muscle mass suffer from the complexity of their environment, but in vitro models are limited because cultured myotubes differ markedly from the mature myofibres of living tissues. We investigate ex vivo maintenance of the isolated myofibre as a model of cellular atrophy. We develop a method, using standard microscopy equipment and widely available analysis software, for measuring f‐actin content per myofibre and per nucleus, and show that the isolated myofibre atrophies at a rate of 35% per week over two weeks of ex vivo maintenance. We characterize early gene expression changes of this model, and use the model to investigate putative modulators of muscle mass, including Myostatin and Follistatin. We establish the isolated myofibre as a tool for the investigation of atrophy, and we present a method of wide applicability for the assay of change in muscle mass at the cellular level. Applying this novel method to the isolated myofibre model we demonstrate ex vivo atrophy of the myofibre. Additional experiments using nuclear markers and the incorporation of a nucleotide analogue (EdU) indicate that the observed atrophy is not associated with loss or replacement of myonuclei. Supported by DoD.