Abstract
The involvement of soluble N-ethylmaleimide sensitive factor-attachment protein receptor (SNARE) proteins in the cAMP-induced exocytosis of aquaporin 2 (AQP2)-containing vesicles was investigated in AQP2-transfected renal CD8 cells. RT-PCR and western blot analysis confirmed the presence of the SNARE homologs VAMP/synaptobrevin-2, syntaxin-1, syntaxin-4 and SNAP-23 in CD8 cells. Tetanus neurotoxin (TeNT) was efficient in cleaving synaptobrevin-like protein both in vitro and in intact CD8 cells incubated with the toxin. TeNT treatment in intact CD8 cells completely abolished cAMP-stimulated AQP2 targeting to the plasma membrane, as assessed by quantification of cell-surface immunoreactivity to an anti-AQP2 antibody raised against a peptide reproducing the extracellular AQP2 C-loop. These results represent the first evidence for the functional involvement of VAMP-2 in cAMP-induced AQP2 exocytosis in renal cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies
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Aquaporin 2
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Aquaporin 6
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Aquaporins / metabolism*
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Cell Membrane / drug effects
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Cell Membrane / immunology
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Cell Membrane / metabolism*
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Cells, Cultured
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Cyclic AMP / metabolism
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Epithelial Cells / cytology
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Epithelial Cells / drug effects
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Epithelial Cells / metabolism
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Exocytosis / drug effects
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Exocytosis / physiology*
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Immunohistochemistry
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Kidney Tubules, Collecting / cytology
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Kidney Tubules, Collecting / metabolism*
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Membrane Proteins / drug effects
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Membrane Proteins / metabolism*
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Metalloendopeptidases
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Models, Biological
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Protein Structure, Tertiary / physiology
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Protein Transport / physiology*
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R-SNARE Proteins
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Rabbits
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Tetanus Toxin
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Transport Vesicles / metabolism*
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Water-Electrolyte Balance / physiology
Substances
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Antibodies
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Aquaporin 2
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Aquaporin 6
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Aquaporins
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Membrane Proteins
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R-SNARE Proteins
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Tetanus Toxin
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tetanospasmin
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Cyclic AMP
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Metalloendopeptidases