DOPAMINE (DA) METABOLISM IN PC12 CELLS EXPOSED TO MANGANESE (MN) AT DIFFERENT OXIDATION-STATES

The present study was aimed at assessing the role of Mn valency state in Mn-induced changes in DA metabolism by PC12 cells. Mn(II)Cl-2, Mn(I II)Acetate, and Mn(IV)O-2 were used for these experiments. PC12 cells were incubated for 3, 24 and 72 hours to Mn nominal concentrations ran ging from 10(-8) to 10(-4) M in 24-well plates containing 2 x 10(5) ce lls/well. Supernatants and cellular materials were then separated and immediately processed for the analysis of dopamine (DA), and its metab olite 3,4-di-hydroxyphenylacetic acid (DOPAC). Lactate dehydrogenase ( LDH) activity and MTT cleavage were measured as indices of cell death. In parallel experiments, Mn-containing medium (10(-5) M) was removed and cells incubated for further periods with Mn-free medium to evaluat e the reversibility of observed changes. At the end of the experimenta l periods, none of Mn-exposed cultures showed appreciable reduction in cell viability as compared to their respective controls. After exposu re to Mn(II) and Mn(III), irreversible and dose-dependent decreases in the medium but not in intra-cellular DA were apparent. Indeed, 10(-4) M Mn(II) caused the disappearance of DA and DOPAC from the medium. Th e same effect was caused by 10(-5) M Mn(III), the dose-effect relation ship being shifted towards lower dose levels. Mn(IV) induced a paralle l and dose-dependent decrease of DA and DOPAC concentrations in both i ntra- and extra-cellular compartments. Such an effect was reversible a fter removal of Mn from the medium. Multiple interferences on DA metab olism are caused by Mn. Mn(II) and Mn(III) seem to block DA secretion without affecting DA turnover rate. Mn(IV) seems to cause DA depletion and aspecific (secondary) changes in secretion rates. Further studies are necessary to understand the mechanisms underlying the differentia l effects of various Mn compounds on DA metabolism. (C) 1996 Inter Pre ss, Inc.