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Single synaptic vesicles fusing transiently and successively without loss of identity


Vesicle fusion and recycling are particularly critical for ongoing neurotransmitter release1,2,3,4 in the small nerve terminals of the brain, which typically contain about 30 functional vesicles4,5. However, the modes of exocytosis and endocytosis that operate at synapses of the central nervous system are incompletely understood. Here we show real-time visualization of a single vesicle fusing at a small synapse of the central nervous system, made possible by highly intensified charge-coupled device imaging of hippocampal synaptic terminals, in which a single vesicle was labelled with the fluorescent membrane marker FM1-43 (ref. 6). In a small number of cases, full loss of fluorescent membrane dye was elicited by a single action potential, consistent with classical complete collapse1. In most cases, however, action potentials triggered only partial loss of fluorescence, suggesting vesicular retention of membrane marker, consistent with ‘kiss-and-run’ vesicle cycling3,4,7,8,9. An alternative hypothesis of independent fusion of partially stained vesicles arising from endosomal splitting could be excluded by observations on the size and timing of successive fusion events. Thus, our experimental evidence supports a predominance of kiss-and-run fusion events10,11,12 and rapid vesicular re-use11.

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Figure 1: Quantal staining of FM1-43 at single boutons.
Figure 2: Single vesicles loaded during minimal stimulation remain in the RRP and retain dye after first fusion.
Figure 3: Individual fusion events usually result in dye retention.
Figure 4: Single vesicles can fuse more than once during 20 action potentials.


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We thank R. W. Aldrich and N. Harata for critically reading the manuscript; S. J. Smith for advice; and members of the Tsien laboratory for comments. This work was supported by grants from the NIMH (R.W.T.) and the Medical Scientist Training Program (A.M.A. and J.L.P.).

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Correspondence to R. W. Tsien.

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Aravanis, A., Pyle, J. & Tsien, R. Single synaptic vesicles fusing transiently and successively without loss of identity. Nature 423, 643–647 (2003).

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