Protocol for electron microscopy ultrastructural localization of the fusogenic lipid phosphatidic acid on plasma membrane sheets from chromaffin cells

Protocol for electron microscopy ultrastructural localization of the fusogenic lipid phosphatidic acid on plasma membrane sheets from chromaffin cells

Summary: 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 p...

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Bibliographic Details
Main Authors: Emeline Tanguy, Tamou Thahouly, Cathy Royer, Valérie Demais, Stéphane Gasman, Sylvette Chasserot-Golaz, Nicolas Vitale
Format: Article
Language:English
Published: Elsevier 2021-06-01
Series:STAR Protocols
Subjects:
Cell biology
Cell membrane
Microscopy
Neuroscience
Structural biology
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721001714
Description
Summary:Summary: 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).
ISSN:2666-1667

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