Loss of MUNC18-1 leads to retrograde transport defects in neurons.
Animals
Biological Transport
/ genetics
Cell Death
Cell Membrane
/ metabolism
Cells, Cultured
Cholera Toxin
/ metabolism
Endoplasmic Reticulum
/ metabolism
Golgi Apparatus
/ pathology
Immunohistochemistry
Intracellular Membranes
/ metabolism
Membrane Glycoproteins
/ genetics
Metabolic Networks and Pathways
/ genetics
Mice
Mice, Knockout
Munc18 Proteins
/ deficiency
Neurodegenerative Diseases
/ genetics
Neurons
/ pathology
SNARE Proteins
/ deficiency
Journal
Journal of neurochemistry
ISSN: 1471-4159
Titre abrégé: J Neurochem
Pays: England
ID NLM: 2985190R
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
revised:
24
11
2020
received:
24
07
2020
accepted:
26
11
2020
pubmed:
2
12
2020
medline:
22
6
2021
entrez:
1
12
2020
Statut:
ppublish
Résumé
Loss of the exocytic Sec1/MUNC18 protein MUNC18-1 or its target-SNARE partners SNAP25 and syntaxin-1 results in rapid, cell-autonomous and unexplained neurodegeneration, which is independent of their known role in synaptic vesicle exocytosis. cis-Golgi abnormalities are the earliest cellular phenotypes before degeneration occurs. Here, we investigated whether loss of MUNC18-1 causes defects in intracellular membrane transport pathways in primary murine neurons that may explain neurodegeneration. Electron, confocal and super resolution microscopy confirmed that loss of MUNC18-1 expression results in a smaller cis-Golgi. In addition, we now show that medial-Golgi and the trans-Golgi Network are also affected. However, stacking and cisternae ultrastructure of the Golgi were normal. Overall, ultrastructure of null mutant neurons was remarkably normal just hours before cell death occurred. By synchronizing protein trafficking by conditional cargo retention in the endoplasmic reticulum using selective hooks (RUSH) and immunocytochemistry, we show that anterograde Endoplasmic Reticulum-to-Golgi and Golgi exit of endogenous and exogenous proteins were normal. In contrast, loss of MUNC18-1 caused reduced retrograde Cholera Toxin B-subunit transport from the plasma membrane to the Golgi. In addition, MUNC18-1-deficiency resulted in abnormalities in retrograde TrkB trafficking in an antibody uptake assay. We conclude that MUNC18-1 deficient neurons have normal anterograde but reduced retrograde transport to the Golgi. The impairments in retrograde pathways suggest a role of MUNC18-1 in endosomal SNARE-dependent fusion and provide a plausible explanation for the observed Golgi abnormalities and cell death in MUNC18-1 deficient neurons.
Identifiants
pubmed: 33259669
doi: 10.1111/jnc.15256
pmc: PMC8247427
doi:
Substances chimiques
Membrane Glycoproteins
0
Munc18 Proteins
0
SNARE Proteins
0
Stxbp1 protein, mouse
0
Tgoln2 protein, mouse
0
Cholera Toxin
9012-63-9
Banques de données
RefSeq
['15256', 'RRID:AB_398141', 'RRID:AB_449893', 'RRID:AB_1524486', 'RRID:AB_310445', 'RRID:AB_2077527', 'RRID:AB_2721247', 'RRID:AB_371415', 'RRID:AB_2096814']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
450-466Informations de copyright
© 2020 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry.
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