Visualization of α-synuclein trafficking via nanogold labeling and electron microscopy.

alpha-Synuclein Microscopy, Electron Cells, Cultured Neurons
Cell culture Microscopy Molecular/Chemical Probes Neuroscience

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
17 03 2023
Historique:
received: 20 09 2022
revised: 09 12 2022
accepted: 24 01 2023
medline: 23 10 2023
pubmed: 3 3 2023
entrez: 2 3 2023
Statut: ppublish

Résumé

There is conflicting evidence regarding the mechanisms of α-synuclein internalization, and its trafficking itinerary following cellular entry remains largely unknown. To examine these issues, we describe steps for coupling α-synuclein preformed fibrils (PFFs) to nanogold beads and their subsequent characterization by electron microscopy (EM). Then we describe the uptake of conjugated PFFs by U2OS cells plated on Permanox 8-well chamber slides. This process eliminates the reliance on antibody specificity and the need to employ complex immunoEM staining protocols. For complete details on the use and execution of this protocol, please refer to Bayati et al. (2022).

Identifiants

DOI: 10.1016/j.xpro.2023.102113 PMID: 36861831 PMC: PMC9988663
pubmed: 36861831
pii: S2666-1667(23)00071-0
doi: 10.1016/j.xpro.2023.102113
pmc: PMC9988663
pii:
doi:

Substances chimiques

alpha-Synuclein 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

102113

Informations de copyright

Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Armin Bayati (A)

Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, QC H3A 2B4, Canada. Electronic address: armin.bayati@mail.mcgill.ca.

Wen Luo (W)

The Neuro's Early Drug Discovery Unit (EDDU), McGill University, Montreal, QC H3A 2B4, Canada.

Esther Del Cid-Pellitero (E)

Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, QC H3A 2B4, Canada.

Edward A Fon (EA)

Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, QC H3A 2B4, Canada.

Thomas M Durcan (TM)

The Neuro's Early Drug Discovery Unit (EDDU), McGill University, Montreal, QC H3A 2B4, Canada. Electronic address: thomas.durcan@mcgill.ca.

Peter S McPherson (PS)

Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, QC H3A 2B4, Canada. Electronic address: peter.mcpherson@mcgill.ca.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines associées à la maladie de Parkinson › alpha-Synucléine › Visualization of α-synuclein trafficking via...
questionsmedicales.fr › Techniques d'investigation › Microscopie électronique › Visualization of α-synuclein trafficking via...
questionsmedicales.fr › Cellules › Cellules cultivées › Visualization of α-synuclein trafficking via...
questionsmedicales.fr › Cellules › Neurones › Visualization of α-synuclein trafficking via...