Measurements of Compensatory Endocytosis by Antibody Internalization and Quantification of Endocytic Vesicle Distribution in Adrenal Chromaffin Cells.
Fiche publication
Date publication
janvier 2021
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr GASMAN Stéphane, Dr CHASSEROT-GOLAZ Sylvette, Dr VITALE Nicolas, Dr ORY Stéphane
Tous les auteurs :
Ceridono M, Chasserot-Golaz S, Vitale N, Gasman S, Ory S
Lien Pubmed
Résumé
Plasma membrane proteins are amenable to endocytosis assays since they are easily labeled by reagents applied in the extracellular medium. This has been widely exploited to study constitutive endocytosis or ligand-induced receptor endocytosis. Compensatory endocytosis is the mechanism by which components of secretory vesicles are retrieved after vesicle fusion with the plasma membrane in response to cell stimulation and a rise in intracellular calcium. Luminal membrane proteins from secretory vesicles are therefore transiently exposed at the plasma membrane. Here, we described an antibody-based method to monitor compensatory endocytosis in chromaffin cells and present an image-based analysis to quantify endocytic vesicles distribution.
Mots clés
Chromaffin cells, Compensatory endocytosis, Dopamine-β-hydroxylase, Internalization, Large dense core vesicle, Neuroendocrine cells
Référence
Methods Mol Biol. 2021 ;2233:43-51