The PKCε activator 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA) stimulates vesicular transport of α7 ACh receptor. The present study was conducted to explore the underlying mechanism using rat hippocampal slices. DCP-LA enhanced serine phosphorylation of the motor protein myosin Va, that moves on the actin filament towards the plus-end (the plasma membrane) and carries cargos, and the effect was abrogated by the PKC inhibitor GF109203X. DCP-LA increased an association of N-ethylmaleimide-sensitive factor (NSF) and myosin Va, but not myosin VI, in a PKC-dependent manner. Moreover, DCP-LA increased an association of myosin Va and α7 ACh receptor in a PKC-dependent manner. Overall, the results of the present study indicate that PKCε, activated by DCP-LA, phosphorylates myosin Va, to increase assembly of NSF/myosinVa/α7 ACh receptor. This may account for DCP-LA-induced stimulation of vesicular transport and exocytosis of α7 ACh receptor.