Ischemia induces early changes to cytoskeletal and contractile proteins in diseased human myocardium.
Ischemia is known to produce damage to subcellular organelles, such as nuclei and mitochondria, in myocardial tissue. We tested the hypothesis that during myocardial ischemia various cytoskeletal and contractile proteins also undergo changes. We induced total global ischemia by incubation in buffer of tissue samples from six human left ventricles that were obtained from heart transplant recipients. Samples were removed from the incubation medium at different time intervals and investigated by immunohistochemistry using monoclonal antibodies against myosin, actin, tropomyosin, troponin T, myomesin, desmin, tubulin, and vinculin. The degree of ischemic injury was determined by electron microscopy. Ischemic cardiomyopathic human tissue showed disturbances of the localization pattern of myosin, actin, tropomyosin, and troponin T as early as 10 minutes after the onset of ischemia; this disruption was complete at 20 minutes. Tubulin also started changing at 10 minutes, but complete disruption was only evident after 120 minutes. Desmin and myomesin showed an intermediate response; changes began at 30 to 40 minutes, and disruption was complete at 90 to 120 minutes. Vinculin was most resistant to ischemia. Ultrastructurally, the tissue showed moderate reversible ischemic injury during the entire period of 180 minutes. Measuring the exposure time in seconds allowed quantitation of the intensity of the fluorescence. We reached the following conclusions: (1) Ischemia causes damage to the contractile proteins sooner than to the cytoskeleton and subcellular organelles. (2) Diseased human hearts are extremely susceptible to the effects of ischemia. These findings are important for the situation of induced cardiac arrest in heart operations and for preservation of donor hearts for transplantation.
Hein
S
S
Max-Planck-Institute for Physiological and Clinical Research, Department of Experimental Cardiology, Bad Nauheim, Germany.
Scheffold
T
T
Schaper
J
J
D000199:Actins / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D002467:Cell Nucleus / Q000648:ultrastructure; D064211:Connectin; D003285:Contractile Proteins / Q000648:ultrastructure*; D003599:Cytoskeleton / Q000648:ultrastructure*; D003893:Desmin / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009124:Muscle Proteins / Q000032:analysis; D017202:Myocardial Ischemia / Q000473:pathology*; D009206:Myocardium / Q000648:ultrastructure*; D009218:Myosins / Q000032:analysis; D015388:Organelles / Q000648:ultrastructure*; D014335:Tropomyosin / Q000032:analysis; D014336:Troponin / Q000032:analysis; D020107:Troponin T; D014404:Tubulin / Q000032:analysis; D016596:Vinculin / Q000032:analysis
D000199:Actins; D015415:Biomarkers; D064211:Connectin; D003285:Contractile Proteins; D003893:Desmin; D009124:Muscle Proteins; D014335:Tropomyosin; D014336:Troponin; D020107:Troponin T; D014404:Tubulin; D016596:Vinculin; D009218:Myosins