Tetanus insensitive VAMP2 differentially restores synaptic and dense core vesicle fusion in tetanus neurotoxin treated neurons.
Animals
Cells, Cultured
Cerebral Cortex
/ cytology
Exocytosis
/ drug effects
Genes, Reporter
Membrane Fusion
/ drug effects
Metalloendopeptidases
Mice
Nerve Tissue Proteins
/ drug effects
Neurons
/ drug effects
Neuropeptide Y
/ analysis
Recombinant Proteins
/ metabolism
Secretory Vesicles
/ drug effects
Synaptic Vesicles
/ drug effects
Tetanus Toxin
/ pharmacology
Vesicle-Associated Membrane Protein 2
/ drug effects
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
02 07 2020
02 07 2020
Historique:
received:
11
11
2019
accepted:
11
06
2020
entrez:
4
7
2020
pubmed:
4
7
2020
medline:
18
12
2020
Statut:
epublish
Résumé
The SNARE proteins involved in the secretion of neuromodulators from dense core vesicles (DCVs) in mammalian neurons are still poorly characterized. Here we use tetanus neurotoxin (TeNT) light chain, which cleaves VAMP1, 2 and 3, to study DCV fusion in hippocampal neurons and compare the effects on DCV fusion to those on synaptic vesicle (SV) fusion. Both DCV and SV fusion were abolished upon TeNT expression. Expression of tetanus insensitive (TI)-VAMP2 restored SV fusion in the presence of TeNT, but not DCV fusion. Expression of TI-VAMP1 or TI-VAMP3 also failed to restore DCV fusion. Co-transport assays revealed that both TI-VAMP1 and TI-VAMP2 are targeted to DCVs and travel together with DCVs in neurons. Furthermore, expression of the TeNT-cleaved VAMP2 fragment or a protease defective TeNT in wild type neurons did not affect DCV fusion and therefore cannot explain the lack of rescue of DCV fusion by TI-VAMP2. Finally, to test if two different VAMPs might both be required in the DCV secretory pathway, Vamp1 null mutants were tested. However, VAMP1 deficiency did not reduce DCV fusion. In conclusion, TeNT treatment combined with TI-VAMP2 expression differentially affects the two main regulated secretory pathways: while SV fusion is normal, DCV fusion is absent.
Identifiants
pubmed: 32616842
doi: 10.1038/s41598-020-67988-2
pii: 10.1038/s41598-020-67988-2
pmc: PMC7331729
doi:
Substances chimiques
Nerve Tissue Proteins
0
Neuropeptide Y
0
Recombinant Proteins
0
Tetanus Toxin
0
Vesicle-Associated Membrane Protein 2
0
vesicle-associated membrane protein 2, mouse
0
Metalloendopeptidases
EC 3.4.24.-
zinc-endopeptidase, tetanus neurotoxin
EC 3.4.24.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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