Journal
MUSCLE & NERVE
Volume 55, Issue 6, Pages 902-912Publisher
WILEY
DOI: 10.1002/mus.25423
Keywords
fiber type; humans; microdissection; muscle weakness; quadriceps muscle
Categories
Funding
- Technology Strategy Board (TSB)
- NIHR Respiratory Biomedical Research Unit at the Royal Brompton and Harefield NHS Foundation Trust and Imperial College
- British Heart Foundation [FS/12/8/29377] Funding Source: researchfish
- Medical Research Council [MR/J000620/1, G0701628] Funding Source: researchfish
- National Institute for Health Research [NF-SI-0515-10093] Funding Source: researchfish
- MRC [G0701628, MR/J000620/1] Funding Source: UKRI
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Introduction: Quadriceps dysfunction is important in chronic obstructive pulmonary disease (COPD), with an associated increased proportion of type II fibers. Investigation of protein synthesis and degradation has yielded conflicting results, possibly due to study of whole biopsy samples, whereas signaling may be fiber-specific. Our objective was to develop a method for fiber-specific gene expression analysis. Methods: 12 COPD and 6 healthy subjects underwent quadriceps biopsy. Cryosections were immunostained for type II fibers, which were separated using laser capture microdissection (LCM). Whole muscle and different fiber populations were subject to quantitative polymerase chain reaction. Results: Levels of muscle-RING-finger-protein-1 and Atrogin-1 were lower in type II fibers of COPD versus healthy subjects (P=0.02 and P=0.03, respectively), but differences were not apparent in whole muscle or type I fibers. Conclusions: We describe a novel method for studying fiber-specific gene expression in optimum cutting temperature compound-embedded muscle specimens. LCM offers a more sensitive way to identify molecular changes in COPD muscle.
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