Abstract
A key question in understanding mechanisms of neurotransmitter release is whether the fusion pore of a synaptic vesicle regulates the amount of transmitter released during exocytosis. We measured dopamine release from small synaptic vesicles of rat cultured ventral midbrain neurons using carbon fiber amperometry. Our data indicate that small synaptic vesicle fusion pores flicker either once or multiple times in rapid succession, with each flicker releasing ∼25–30% of vesicular dopamine. The incidence of events with multiple flickers was reciprocally regulated by phorbol esters and staurosporine. Thus, dopamine neurons regulate the amount of neurotransmitter released by small synaptic vesicles by controlling the number of fusion pore flickers per exocytotic event. This mode of exocytosis is a potential mechanism whereby neurons can rapidly reuse vesicles without undergoing the comparatively slow process of recycling.
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Acknowledgements
We thank Q. Al-Awqati, K. Larsen, M. Nirenberg and Y. Schmitz for critique of the manuscript, and A. Petrenko for α-latrotoxin. Supported by the National Alliance for Research on Schizophrenia and Depression, the Lowenstein Foundation, the Parkinson's Disease Foundation, the National Institute on Drug Abuse and the National Institute of Neurological Disorders and Stroke.
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Supplementary Fig. 1
To estimate the frequency of stochastically overlapping events, the fraction of interspike intervals within 1 s bins is indicated on the y axis. The best-fit exponential decay and corresponding time constant determined by least squares is shown for each experimental group. (JPG 32 kb)
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Staal, R., Mosharov, E. & Sulzer, D. Dopamine neurons release transmitter via a flickering fusion pore. Nat Neurosci 7, 341–346 (2004). https://doi.org/10.1038/nn1205
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DOI: https://doi.org/10.1038/nn1205
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