MPP+ downregulates mitochondrially encoded gene transcripts and their activities in dopaminergic neuronal cells: Protective role of Bcl-2

The effects of neurotoxins on levels of mitochondrially encoded gene transc ripts in a dopaminergic neuronal cell line, MN9D, were examined following t reatment with 200 muM N-methyl-4-phenylpyridinium (MPP+) or 6-hydroxydopami ne (6-OHDA). As confirmed by a Northern blot analysis, levels of cytochrome c oxidase subunit 3 (COX III) and ATPase subunit 6 (ATPase 6) transcript w ere decreased in a time-dependent manner following treatment with MPP+ but not with 6-OHDA. Accordingly, enzymatic activity of cytochrome c oxidase (C OX) and the intracellular ATP content were also decreased in MPP+-treated c ells while these remained unaltered in 6-OHDA-treated cells. In the cell de ath paradigm induced by MPP+, overexpression of Bcl-2 in MN9D cells (MN9D/B cl-2) significantly blocked MPP+-induced downregulation of COX III and ATPa se 6 transcripts. In MN9D/Bcl-2 cells, MPP+-induced downregulation of COX a ctivity and the intracellular level of ATP was also blocked. Treatment with a pan-caspase inhibitor, however, neither prevented MPP+-induced downregul ation of COX activity nor affected intracellular level of ATP in MN9D cells . Taken together, our present data suggest that Bcl-2 may play a regulatory role in energy metabolism by preventing downregulation of mitochondrially encoded gene(s) at a point distinct from its known anticaspase activity in MPP+-induced dopaminergic neuronal death. (C) 2001 Academic Press.