Mitochondrial dysfunction has been associated with Parkinson's disease. However, the role of mitochondrial defects in the formation of Lewy bodies, a pathological hallmark of Parkinson's disease has not been addressed directly. In this report, we investigated the effects of inhibitors of the mitochondrial electron-transport chain on the aggregation of ␣-synuclein, a major protein component of Lewy bodies. Treatment with rotenone, an inhibitor of complex I, resulted in an increase of detergent-resistant ␣-synuclein aggregates and a reduction in ATP level. Another inhibitor of the electron-transport chain, oligomycin, also showed temporal correlation between the formation of aggregates and ATP reduction. Microscopic analyses showed a progressive evolution of small aggregates of ␣-synuclein to a large perinuclear inclusion body. The inclusions were co-stained with ubiquitin, 20 S proteasome, ␥-tubulin, and vimentin. The perinuclear inclusion bodies, but not the small cytoplasmic aggregates, were thioflavin S-positive, suggesting the amyloid-like conformation. Interestingly, the aggregates disappeared when the cells were replenished with inhibitor-free medium. Disappearance of aggregates coincided with the recovery of mitochondrial metabolism and was partially inhibited by proteasome inhibitors. These results suggest that the formation of ␣-synuclein inclusions could be initiated by an impaired mitochondrial function and be reversed by restoring normal mitochondrial metabolism.Many neurodegenerative diseases are associated with characteristic intracellular protein inclusions in the affected brain regions (1, 2). Lewy body (LB) 1 is one of the inclusions that is associated with Parkinson's disease (PD) and related neurologic diseases, including dementia with Lewy bodies and Lewy body variant of Alzheimer's disease (3). Aggregation of ␣-synuclein (␣-syn) appears to be essential for the LB formation since fibrillar aggregates of ␣-syn are major components of LBs (4, 5), thus the diseases that are characterized by LBs are collectively referred to as ␣-synucleinopathies. ␣-Syn itself forms typical amyloids in solution by a nucleation-dependent mechanism (6 -8). Importantly, two autosomal dominant mutations of the ␣-syn gene were linked to rare familial earlyonset PD (9, 10), and the mutant proteins tend to aggregate more rapidly than the wild type (11-13). Transgenic flies modified to express human ␣-syn at various levels develop age-dependent pathological and behavioral changes that resemble human PD, including LB-like intraneuronal inclusions, loss of dopaminergic neurons, and a decline in locomotor activity (14). Also, transgenic mice that express human ␣-syn produced neuronal inclusions and dopaminergic presynaptic degeneration (15). In addition, the extent of formation of these LB-like inclusions has been shown to correlate with the expression level of ␣-syn in transgenic animals (15). These findings provide compelling evidence that the aggregation of ␣-syn is directly linked to LB formation, and therefore to the ...