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The presynaptic scaffolding protein Piccolo organizes the readily releasable pool at the calyx of Held

Bassoon and Piccolo do not mediate basal synaptic vesicle release at a high‐frequency synapse. Knockdown of Bassoon increases short‐term depression at the calyx of Held. Both Bassoon and Piccolo have shared functions in synaptic vesicle replenishment during high‐frequency synaptic transmission. Piccolo organizes the readily releasable pool of synaptic vesicles. It safeguards a fraction of them to be not immediately available for action potential‐induced release. This enables the synapse to sustain high‐frequency synaptic transmission over long periods.