Synaptic vesicles (SVs) play a key role in synaptic transmission. This study used time-lapse microscopy to demonstrate two distinct mechanisms of vesicle traffic along axons. They utilized specific pharmacological interventions along with imaging of transient actin polymerization, and traffic of recycling SVs loaded with single quantum dots to characterize the underlying axonal actin structures. For this study, hippocampal neurons were cultured from Sprague-Dawley rat pups. The dataset contains time-lapse imaging of vesicle traffic and actin polymerization, and immunocytochemistry data. This data indicate that the axonal traffic of recycling vesicles is not supported by ubiquitous microtubule-based motility, but relies on actin filaments instead.