Protocol for electron microscopy ultrastructural localization of the fusogenic lipid phosphatidic acid on plasma membrane sheets from chromaffin cells.
Cell biology
Cell membrane
Microscopy
Neuroscience
Structural biology
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
18 06 2021
18 06 2021
Historique:
entrez:
29
4
2021
pubmed:
30
4
2021
medline:
30
4
2021
Statut:
epublish
Résumé
The glycerophospholipid phosphatidic acid (PA) is a key player in regulated exocytosis, but little is known about its localization at the plasma membrane. Here, we provide a protocol for precisely determining the spatial distribution of PA at exocytotic sites by electron microscopy. Using primary bovine chromaffin cells expressing a PA sensor (Spo20p-GFP), we describe the process for cell stimulation and detergent-free preparation of plasma membrane sheets. The protocol can be applied to other cell models and to distinct membrane lipids. For complete details on the use and execution of this protocol, please refer to Tanguy et al. (2020).
Identifiants
pubmed: 33912850
doi: 10.1016/j.xpro.2021.100464
pii: S2666-1667(21)00171-4
pmc: PMC8065343
doi:
Substances chimiques
Phosphatidic Acids
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Pagination
100464Informations de copyright
© 2021 Centre National pour la Recherche Scientifique.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
Références
Cell Rep. 2020 Aug 18;32(7):108026
pubmed: 32814056
J Cell Biol. 2015 Aug 31;210(5):785-800
pubmed: 26323692
FASEB J. 2020 May;34(5):6769-6790
pubmed: 32227388
Traffic. 2010 Jul 1;11(7):958-71
pubmed: 20374557
J Neurochem. 2016 Jun;137(6):904-12
pubmed: 26877188
Front Cell Neurosci. 2019 Jan 23;13:2
pubmed: 30728767
Handb Exp Pharmacol. 2020;259:115-130
pubmed: 30570690
J Biol Chem. 2007 Jul 27;282(30):21746-57
pubmed: 17540765
J Neurosci. 2013 Feb 20;33(8):3545-56
pubmed: 23426682
J Biol Chem. 2017 Mar 10;292(10):4266-4279
pubmed: 28115519
Methods Mol Biol. 2021;2233:169-179
pubmed: 33222134
Methods Cell Biol. 2012;108:445-59
pubmed: 22325614
Methods Mol Biol. 2021;2233:301-309
pubmed: 33222143