Abstract
We have examined the kinetics by which FM1-43 escapes from individual synaptic vesicles during exocytosis at hippocampal boutons. Two populations of exocytic events were observed; small amplitude events that lose dye slowly, which made up more than half of all events, and faster, larger amplitude events with a fluorescence intensity equivalent to single stained synaptic vesicles. These populations of destaining events are distinct in both brightness and kinetics, suggesting that they result from two distinct modes of exocytosis. Small amplitude events show tightly clustered rate constants of dye release, whereas larger events have a more scattered distribution. Kinetic analysis of the association and dissociation of FM1-43 with membranes, in combination with a simple pore permeation model, indicates that the small, slowly destaining events may be mediated by a narrow approximately 1-nm fusion pore.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Animals, Newborn
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Antibodies, Monoclonal / metabolism
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Calcium-Binding Proteins / metabolism
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Cells, Cultured
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Dose-Response Relationship, Drug
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Exocytosis*
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Fluorescent Dyes* / metabolism
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Fluorescent Dyes* / pharmacology
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Hippocampus / cytology
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Hippocampus / physiology*
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Kinetics
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Liposomes
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Melitten / pharmacology
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Membrane Glycoproteins / metabolism
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Membrane Proteins / metabolism
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Mice
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Models, Biological
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Models, Neurological
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Nerve Tissue Proteins / metabolism
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Precipitin Tests
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Presynaptic Terminals / metabolism
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Pyridinium Compounds* / metabolism
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Pyridinium Compounds* / pharmacology
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Quaternary Ammonium Compounds* / metabolism
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Quaternary Ammonium Compounds* / pharmacology
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R-SNARE Proteins
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Rats
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Receptors, N-Methyl-D-Aspartate / metabolism
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Synapses / physiology*
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Synaptic Vesicles / metabolism*
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Synaptotagmins
Substances
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Antibodies, Monoclonal
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Calcium-Binding Proteins
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FM1 43
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Fluorescent Dyes
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Liposomes
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Membrane Glycoproteins
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Membrane Proteins
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NMDA receptor A1
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Nerve Tissue Proteins
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Pyridinium Compounds
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Quaternary Ammonium Compounds
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R-SNARE Proteins
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Receptors, N-Methyl-D-Aspartate
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Synaptotagmins
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Melitten