Myosin V is an unconventional type of actin-binding myosin that participates in cytoplasmic organelle transport. Although this unconventional myosin has been extensively studied, its subcellular localization in the mammalian cerebellum has not been determined. In this work, we used an antibody against the tail domain of the myosin-Va heavy chain and a secondary antibody labeled with protein A-gold (15 nm) to study the subcellular distribution of this protein. Myosin-Va was found in the cytoplasm, where it was associated with a filament (probably actin). This protein was also detected in the plasma membrane of axons and dendrites in the molecular layer in rat cerebellum.