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1 Medical, Surgical, and Research Services, Philadelphia Veterans Affairs Medical Center, and 2 Departments of Medicine, Surgery, and Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia 19104; and 3 Department of Pediatrics, Magee-Womens Research Institute, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15213
In preliminary experiments we noted developmental (i.e., embryonic and neonatal) myosin heavy chains (MHCs) in the diaphragms of patients with severe chronic obstructive pulmonary disease (COPD). We hypothesized that this finding represented new fiber formation secondary to injury associated with the mechanical stress of COPD or previously undescribed MHCs in the human diaphragm. To distinguish between these possibilities, we analyzed diaphragmatic biopsies obtained from 9 patients with severe COPD (forced expiratory volume in 1 s = 21 ± 2% predicted, residual volume = 283 ± 22% predicted) and 10 age-matched controls. First, using immunocytochemistry with specific monoclonal antibodies, we noted that control diaphragms had greater proportions of fibers expressing embryonic (50 ± 2 vs. 28 ± 3%, P < 0.0001) and neonatal (52 ± 2 vs. 32 ± 3%, P < 0.001) MHCs than COPD diaphragms. Second, SDS-PAGE demonstrated that these developmental MHCs represented only a very small fraction of the diaphragmatic MHC content. Third, the RT-PCR demonstrated mRNA coding for embryonic and neonatal MHCs in COPD and control diaphragms. Last, COPD and control diaphragms exhibited normal histology on light microscopy. We conclude that the presence of developmental MHC isoforms does not indicate new fiber formation in diaphragms of patients with severe COPD. Although these results represent the first systematic description of embryonic and neonatal MHCs in normal adult human diaphragms, their function remains to be elucidated.
human diaphragmatic muscle; embryonic myosin heavy chain; neonatal myosin heavy chain; fiber types; immunocytochemistry
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