Synaptobrevin is essential for fast synaptic-vesicle endocytosis

Abstract

Synaptobrevin-2 (VAMP-2), the major SNARE protein of synaptic vesicles, is required for fast calcium-triggered synaptic-vesicle exocytosis. Here we show that synaptobrevin-2 is also essential for fast synaptic-vesicle endocytosis. We demonstrate that after depletion of the readily releasable vesicle pool, replenishment of the pool is delayed by knockout of synaptobrevin. This delay was not from a loss of vesicles, because the total number of pre-synaptic vesicles, docked vesicles and actively recycling vesicles was unaffected. However, altered shape and size of the vesicles in synaptobrevin-deficient synapses suggests a defect in endocytosis. Consistent with such a defect, the stimulus-dependent endocytosis of horseradish peroxidase and fluorescent FM1-43 were delayed, indicating that fast vesicle endocytosis may normally be nucleated by a SNARE-dependent coat. Thus, synaptobrevin is essential for two fast synapse-specific membrane trafficking reactions: fast exocytosis for neurotransmitter release and fast endocytosis that mediates rapid reuse of synaptic vesicles.

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Figure 1: Delayed replenishment of synaptic readily releasable pool in synaptobrevin-2 knock-out (Syb2-KO) synapses.
Figure 2: Frequency-dependent facilitation in synaptobrevin-2 knock-out synapses.
Figure 3: Comparable pool-size of functional vesicles in synaptobrevin-2 knock-out and wild-type synapses.
Figure 4: Electron microscopy of stimulated and horseradish-peroxidase-labelled synaptobrevin-2 knock-out synaptic terminals.
Figure 5: Impaired endocytosis in synaptobrevin-2 knock-out synapses.

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Acknowledgements

We are grateful to A. Roth and N. Hamlin for technical assistance. This work was supported by the Howard Hughes Medical Institute (F.D., S.S and T.C.S) and the National Institute for Mental Health (grant MH066198 to E.T.K.).

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Correspondence to Thomas C. Südhof or Ege T. Kavalali.

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Deák, F., Schoch, S., Liu, X. et al. Synaptobrevin is essential for fast synaptic-vesicle endocytosis. Nat Cell Biol 6, 1102–1108 (2004). https://doi.org/10.1038/ncb1185

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