Dopamine metabolism alterations in a manganese-treated pheochromocytoma cell line (PC12).
By monitoring dopamine metabolism in rat pheochromocytoma derived PC12 cell cultures during extended treatment with manganese chloride, we assessed the functional changes occurring in a dopaminergic system during the development of manganese-induced damage. Besides eliciting a specific toxic effect on PC12 cells, manganese induced the complete disappearance of extracellular (free) but not intracellular (vesicle stored) dopamine and its metabolite 3,4-dihydroxyphenylacetic acid. This effect was observed also using low manganese concentrations (1 microM) and mainly occurred by non-enzymatic catechol oxidation since it was still evident in a cell free medium and it was fully prevented by ascorbic acid but not by reduced glutathione. The possibility of a mere "non-biological" action was ruled out by the observation of an irreversible effect of manganese as manifested by the cells' apparent inability to release dopamine or 3,4-dihydroxyphenylacetic acid into the culture medium even after complete manganese removal (post-manganese incubation). That a free radical mechanism was not involved in the genesis of this irreversible effect was shown by the fact that neither ascorbic acid, catalase, superoxide dismutase nor glutathione-peroxidase were able to prevent the decrease in catecholamine levels in the "post-manganese" incubation medium when added at the same time as the manganese. The results establish this PC12 cell system as an in vitro model for studying interactions between manganese and catechols and provide a detailed description of the nature of the neurochemical alterations that this heavy metal can induce in a dopaminergic system.
Vescovi
A
A
Laboratory of Cellular Neuropharmacology, National Neurological Institute C. Besta, Milan, Italy.
Facheris
L
L
Zaffaroni
A
A
Malanca
G
G
Parati
E A
EA
D000310:Adrenal Gland Neoplasms; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002712:Chlorides; D003470:Culture Media; D004298:Dopamine / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D017895:Manganese Compounds; D020149:Manganese Poisoning; D010084:Oxidation-Reduction; D010673:Pheochromocytoma; D014407:Tumor Cells, Cultured; D015032:Zinc / Q000633:toxicity
D002712:Chlorides; D003470:Culture Media; D017895:Manganese Compounds; D007770:L-Lactate Dehydrogenase; D015032:Zinc; D001205:Ascorbic Acid; C025340:manganese chloride; D004298:Dopamine