Effect of iron on the stability of macrophage lysosomes.
Mouse peritoneal macrophages exposed to 60 microM Fe3+ in culture showed a pronounced decline in survival rate and an enhanced capacity for lipid peroxidation, as estimated by the formation of malondialdehyde. The iron-exposed cells contain secondary lysosomes loaded with iron as demonstrated by transmission electron microscopy, energy dispersive X-ray micro-analysis and the sulphide-silver method. The macrophages concentrated within their lysosomes the weak base acridine orange (AO) when the latter was added to the medium in a low concentration (1:10(5)). When cells were viewed under continuous illumination with short wave blue light in a fluorescence microscope, AO-exposed cells initially showed a brilliant red granular fluorescence and subsequently a progressive redistribution of AO into the cell sap and the nucleus with resultant green fluorescence. This redistribution of AO was associated with the disappearance of the normal granular reaction product pattern of the lysosomal marker enzyme acid phosphatase (AP), as demonstrated with a Gomori-type reaction, and later with cell death. The redistribution of AO and AP from the lysosomal vacuome to the cell sap under the influence of blue light occurred markedly more rapidly in macrophages containing iron loaded lysosomes than in control cells. The findings indicate that the presence of iron in lysosomes decreases the proton-retaining ability of their bordering membranes, possibly by way of enhanced metal (iron) catalyzed lipid peroxidation. It appears that leakage of AO signifies destabilization of the lysosomal membrane.
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D007501:Iron / Q000494:pharmacology*; D008247:Lysosomes / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects* / Q000648:ultrastructure; D008297:Male; D008566:Membranes / Q000187:drug effects; D051379:Mice; D008854:Microscopy, Electron
D007501:Iron