AbstractEndocytosis at the presynaptic terminal is initiated by Ca2+ influx through voltage‐gated Ca2+ channels. At the Drosophila neuromuscular junction, we demonstrated two components of endocytosis linked to distinct Ca2+ channels. A voltage‐gated Ca2+ channel blocker, (R)‐(+)‐Bay K8644 (R‐BayK), selectively blocked one component (R‐BayK‐sensitive component) without affecting exocytosis, while low concentrations of La3+ preferentially depressed the other component (La3+ ‐sensitive component). In a temperature‐sensitive mutant, shibirets, at non‐permissive temperatures, dynamin clusters were found immunohistochemically at the active zone (AZ) during the R‐BayK‐sensitive endocytosis, while they were detected at the non‐AZ during the La3+‐sensitive endocytosis. Immunostaining of the Ca2+ channel α2δ subunit encoded by straightjacket (stj) was found within the AZ, and a mutation in stj depressed the R‐BayK‐sensitive component but enhanced the La3+ ‐sensitive one, indicating that the α2δ subunit is associated with the R‐BayK‐sensitive Ca2+ channel. Filipin bound to the non‐AZ membrane and inhibited the La3+ ‐sensitive component, but not the R‐BayK‐sensitive one. We concluded that the R‐BayK‐sensitive component of endocytosis occurred at the AZ and termed this AZ endocytosis. We also concluded that the La3+ ‐sensitive component occurred at the non‐AZ and termed this non‐AZ endocytosis. These two types of endocytosis were modulated by various drugs towards opposite directions, indicating that they were differentially regulated. During high‐frequency stimulation, AZ endocytosis operated mainly in the early phase, whereas non‐AZ endocytosis operated in the late phase. Thus, intense synaptic transmission is coordinately maintained by synaptic vesicle recycling initiated by Ca2+ influx through the two types of Ca2+ channel.