Reduction in beta-myosin heavy chains in stunned myocardium as assessed by nondenaturing gel electrophoresis.
Myosin plays a key role in the structure and function of cardiac muscle. Three myosin isoenzymes (V(1), V(2), and V(3)) with different ATPase activities have been identified in mammalian ventricles based on their heavy chain constituents. The relative amount of myosin isoenzymes changes under physiological and pathological conditions. Until now, myosin isoenzymes have frequently been determined using either tube gel (nondenaturing) polyacrylamide gel electrophoresis (PAGE), or gradient or uniform sodium dodecyl sulfate (denaturing) PAGE. Both methods have disadvantages, e.g., a long running time. We developed, therefore, a uniform, nondenaturing PAGE with slab minigel format for analyzing the myosin isoenzymes in normoxic and stunned rabbit hearts. In normoxic hearts of adult rabbits, V(3) predominated over V(1) (46 vs 41%). In turn, in the stunned hearts, V(1) predominated over V(3) (70 vs 30%), and the heterodimeric V(2) was not anymore detectable. This alteration appears to result from a selective loss of myosin heavy chain (MHC)-beta. In parallel, the biochemical markers troponin I and creatine kinase were increased in the stunned hearts. We suggest that alterations of myosin isoenzymes in stunned myocardium can be monitored with native PAGE. The present analysis of myosin isoenzyme appears thus as a new tool for evaluating defects in MHC dimer formation in postischemic hearts.
Garcia
S C
SC
Department of Surgery I, University Hospital Duesseldorf, Heinrich-Heine-University, Moorenstrasse 5, 40225, Duesseldorf, Germany. sgarpom@smail.ufsm.br
Pomblum
V J
VJ
Gams
E
E
Rupp
H
H
Schipke
J D
JD
D000818:Animals; D015415:Biomarkers; D003402:Creatine Kinase / Q000032:analysis; D003627:Data Interpretation, Statistical; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008297:Male; D017682:Myocardial Stunning / Q000378:metabolism*; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000032:analysis* / Q000378:metabolism*; D011817:Rabbits; D019210:Troponin I / Q000032:analysis
D015415:Biomarkers; D007527:Isoenzymes; D019210:Troponin I; D003402:Creatine Kinase; D018995:Myosin Heavy Chains