ABSTRACT
Mouse embryonic fibroblasts (MEF) with point mutation in somatic cytochrome C gene were generated and characterized. It was shown that substitution of lysine for tryptophan in position 72 (K72W) decreased the proapoptotic functions of cytochrome C in response to staurosporin treatment without disrupting its respiratory functions. The presence of this mutation did not affect the pattern of cytochrome C gene expression or its localization inside the cell. These cell cultures therefore represent an interesting model for study apoptotic signaling and physiological functions of cytochrome C.
Subject(s)
Apoptosis/genetics , Cytochromes c/genetics , Embryo, Mammalian/enzymology , Fibroblasts/enzymology , Models, Biological , Mutation, Missense , Amino Acid Substitution , Animals , Apoptosis/drug effects , Cell Line , Cytochromes c/antagonists & inhibitors , Cytochromes c/metabolism , Embryo, Mammalian/cytology , Enzyme Inhibitors/pharmacology , Fibroblasts/cytology , Mice , Staurosporine/pharmacologyABSTRACT
Experiments on 10 rats and 10 rabbits were made to investigate metabolic aftereffects of 40-minute heat ischemia and subsequent reperfusion. It was found that mitochondrial function deteriorated significantly in an early postischemic period. The disorder manifested with a relative prevalence of cell ATP consumption over its synthesis. This is accompanied with intensive production by mitochondria of nitric oxide and oxygen free radicals. Fluorescent probes and confocal microscopy of vital renal sections showed that mitochondria are responsible for excessive generation of nitric oxide and oxygen radicals in the kidney in an early reperfusion period. The discussion concerns the role of nitric oxide in reperfusion renal damage and participation of mitochondria in formation of its anti-ischemic resistance.
