Permeability of rat heart myocytes to cytochrome c.
Rat heart myocytes undergoing progressive damage demonstrate morphological changes of shortening and swelling followed by the formation of intracellular vacuoles and plasma membrane blebbing. The damaged myocytes displayed impaired N,N'-tetramethyl-p-phenyldiamine (TMPD) ascorbate-stimulated respiratory activity which was restored by the addition of reduced cytochrome c to the cell culture medium. To clarify the role played by cytochrome c in the impairment of cell respiration, polarographic, spectrophotometric and fluorescence as well as electron microscopy imaging experiments were performed. TMPD/ascorbate-stimulated respiratory activity returned to control levels, at approximately 20 microM cytochrome c, establishing the threshold below which the turnover rate by cytochrome c oxidase in the cell depends on cytochrome concentration. Mildly damaged cardiac myocytes, as indicated by cell shortening, retention of visible striations and free-fluorescein exclusion, together with the absence of lactate dehydrogenase leakage and exclusion of trypan blue, were able to oxidize exogenous cytochrome c and were permeable to fluorescein-conjugated cytochrome c. The results, while consistent with an early cytochrome c release observed at the beginning of cell death, elucidate the role played by cytochrome c in the kinetic control of mitochondrial electron transfer under pathological conditions, particularly those involving the terminal part of the respiratory chain. These data are the first to demonstrate that the sarcolemma of cardiac myocytes, damaged but still viable, is permeable to cytochrome c.
Sarti
P
P
Department of Biochemical Sciences, University of Rome La Sapienza, Italy. sarti@axrma.uniroma1.it
Silver
R B
RB
Paroli
L
L
Nikonorov
I
I
Blanck
T J
TJ
D000818:Animals; D016923:Cell Death; D002463:Cell Membrane Permeability; D019069:Cell Respiration; D048429:Cell Size; D002478:Cells, Cultured; D016922:Cellular Senescence; D003574:Cytochrome c Group / Q000378:metabolism*; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism; D007700:Kinetics; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008929:Mitochondria, Heart / Q000201:enzymology / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D010101:Oxygen Consumption; D011048:Polarography; D051381:Rats; D017207:Rats, Sprague-Dawley; D012508:Sarcolemma / Q000378:metabolism / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism
D003574:Cytochrome c Group; D003576:Electron Transport Complex IV