Continuous vesicle cycling in the synaptic terminal of retinal bipolar cells

Endocytosis and exocytosis were investigated in the synaptic terminal of retinal bipolar cells by monitoring the uptake and loss of the fluorescent dye FM1-43. Depolarization in the presence of Ca2+ stimulated a continuous cycle of exocytosis and endocytosis that was approximately balanced at rates up to 3800 vesicles per s. Vesicles became available for exocytosis within 1 min of endocytosis, and about 700,000 releasable vesicles were specifically localized to a region within 2 mu m of the plasma membrane. Release of caged Ca2+ using NP-EGTA while simultaneously monitoring cytosolic Ca2+ with Fura-2 indicated that continuous exocytosis was stimulated by sub-micromolar levels of Ca2+. It has been suggested that the ribbon synapse of bipolar cells only supports transient exocytosis, but our results demonstrate that this synapse is specialized for the continuous secretion of neurotransmitter.