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PMID: 10899113 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Munc13-1 acts as a priming factor for large dense-core vesicles in bovine chromaffin cells.

The EMBO journal ·Vol. 19 ·No. 14 ·2000-07-17 ·Pages 3586-96

Ashery U, Varoqueaux F, Voets T, Betz A, Thakur P, Koch H, Neher E, Brose N, Rettig J

Abstract

In chromaffin cells the number of large dense-core vesicles (LDCVs) which can be released by brief, intense stimuli represents only a small fraction of the 'morphologically docked' vesicles at the plasma membrane. Recently, it was shown that Munc13-1 is essential for a post-docking step of synaptic vesicle fusion. To investigate the role of Munc13-1 in LDCV exocytosis, we overexpressed Munc13-1 in chromaffin cells and stimulated secretion by flash photolysis of caged calcium. Both components of the exocytotic burst, which represent the fusion of release-competent vesicles, were increased by a factor of three. The sustained component, which represents vesicle maturation and subsequent fusion, was increased by the same factor. The response to a second flash, however, was greatly reduced, indicating a depletion of release-competent vesicles. Since there was no apparent change in the number of docked vesicles, we conclude that Munc13-1 acts as a priming factor by accelerating the rate constant of vesicle transfer from a pool of docked, but unprimed vesicles to a pool of release-competent, primed vesicles.

MeSH Terms
Animals Brain Calcium/metabolism Catecholamines/metabolism Cattle Cells, Cultured Chromaffin Cells/cytology,metabolism Computer Simulation Egtazic Acid/analogs & derivatives,metabolism Electric Conductivity Exocytosis Kinetics Membrane Fusion Microscopy, Electron Models, Biological Nerve Tissue Proteins/genetics,metabolism Patch-Clamp Techniques Photolysis Protein Isoforms/genetics,metabolism Rats Synaptic Vesicles/metabolism Synaptosomes/metabolism Vacuoles/metabolism
Chemicals
Catecholamines Nerve Tissue Proteins Protein Isoforms Unc13a protein, rat 2-nitrophenyl-EGTA Egtazic Acid Calcium
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Ashery U
Max-Planck-Institute for Biophysical Chemistry, Department of Membrane Biophysics, Am Fassberg 11, 37077 Göttingen, Germany.
Varoqueaux F
Voets T
Betz A
Thakur P
Koch H
Neher E
Brose N
Rettig J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.14.3586
Published
2000-07-17
Pages
3586-96
Language
English
Region
England
NLM ID
8208664
PMCID
PMC313963
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/19.14.3586 PMC Full Text View journal details
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