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Significance Using our recently designed microfluidic setup, we investigated the early stage of SNAREpin-induced fusion. We discovered the existence of subsecond transient fusion pores with a well-defined subnanometer size that occur when one or two SNAREpins are mediating vesicle fusion. In contrast, when vesicle fusion is mediated by three SNAREpins, the fusion pore reaches a diameter larger than 1.5 nm and expands spontaneously and indefinitely. These results quantitatively explain the need for a complex machinery to ensure a submillisecond neurotransmitter release after the arrival of the action potential during synaptic transmission.

Nascent fusion pore opening monitored at single-SNAREpin resolution