Dynamic microstructure of mitochondrial membranes from rabbit heart subjected to reperfusion after ischemia

The effects of ischemia combined with reperfusion on the dynamic microstructure of mitochondrial membranes were studied in the hearts of anesthetized open-chest rabbits by means of time-resolved fluorometry. The fluorescence of diphenyl hexatriene (DPH) and its anisotropic decay were used to calculate membrane viscosity, wobbling angle of phospholipids and fluorescence life time. The anterior descending branch of the coronary artery was occluded for 15 min and reperfused for 15 min. Mitochondria from the area of the left ventricular wall exposed to ischemia-reperfusion and from an unexposed control area were separately isolated in 8 rabbits. For comparison, mitochondria from ventricular wall exposed to ischemia but not to reperfusion were obtained from 5 rabbits. The membrane viscosity increased from 0.44 to 0.49 poise and the wobbling angle of phospholipids tended to decrease from 59 to 56 degrees at 37 degrees C. The amount of peroxidized lipid rose from 1.5 to 4.3 nmol/mg-protein as expressed with MDA in the mitochondria exposed to ischemia-reperfusion. The fluorescence life time of DPH was slightly longer in the latter than in mitochondria from the control area. None of these parameters was altered in mitochondria exposed only to ischemia. The increase in viscosity observed in mitochondria from the ischemic-reperfused area probably produces a decrease in the diffusion of molecules through mitochondrial membranes. The concurrent tendency of the decrease in wobbling angle of membrane phospholipids suggests a decrease in the size of molecules which can permeate membranes. Limitations in diffusion will affect the metabolic activity of mitochondria and finally, overall cardiac function.