The aggregation of α-synuclein (α-syn) is believed to play a critical role in the pathogenesis of disorders such as dementia with Lewy bodies and Parkinson's disease. The function of α-syn remains unclear, although several lines of evidence suggest that α-syn is involved in synaptic vesicle trafficking, probably via lipid binding, and interactions with lipids have been shown to regulate α-syn aggregation. In this context, the main objective of this study was to determine whether methyl-β-cyclodextrin (MβCD), a cholesterol-extracting agent, interfered with α-syn accumulation in models of synucleinopathy. For this purpose, we studied the effects of MβCD on the accumulation of α-syn in a transfected neuronal cell line and in transgenic mice. Immunoblot analysis showed that MβCD reduced the level of α-syn in the membrane fraction and detergent-insoluble fraction of transfected cells. In agreement with the in vitro studies, treatment of mice with MβCD resulted in decreased levels of α-syn in membrane fractions and reduced accumulation of α-syn in the neuronal cell body and synapses. Taken together, these results suggest that changes in cholesterol and lipid composition using cholesterol-lowering agents may be used as a tool for the treatment of synucleinopathies.