Kinetics and regulation of fast endocytosis at hippocampal synapses

Presynaptic nerve terminals often contain as few as a hundred vesicles(1,2) and so must recycle them soon after exocytosis to preserve synaptic transmission and presynaptic morphology(3,4) during repetitive firing. The kinetics(3,4) and mechanisms(5) of vesicular endocytosis and repriming have therefore been studied. Vesicles in hippocampal nerve terminals can become available to release their contents within similar to 40 s of the previous round of exocytosis(6,7). Studies using the styryl dye FM1-43 (ref. 3) have estimated the time constant for endocytosis as similar to 20-30 s (refs 4, 8), at least half of the total recycling time, which is much slower than endocytosis in other secretory systems(9-11). It seems paradoxical that the neurosecretory terminals that could benefit the most from rapid endocytosis do not use such a mechanism. Here we demonstrate the existence of fast endocytosis in hippocampal nerve terminals and derive its kinetics from fluorescence measurements using dyes with varying rates of membrane departitioning. The rapid mode of vesicular retrieval was much faster after exposure to staurosporine or elevated extracellular calcium. Thus hippocampal synapses take advantage of efficient mechanisms for endocytosis, and their vesicular retrieval is subject to modulatory control.