Flow Cytometric Monitoring of Dynamic Cytosolic Calcium, Sodium, and Potassium Fluxes Following Platelet Activation.

Détails

ID Serval
serval:BIB_DC6551A7EA2B
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Flow Cytometric Monitoring of Dynamic Cytosolic Calcium, Sodium, and Potassium Fluxes Following Platelet Activation.
Périodique
Cytometry. Part A
Auteur(s)
Aliotta A., Bertaggia Calderara D., Alberio L.
ISSN
1552-4930 (Electronic)
ISSN-L
1552-4922
Statut éditorial
Publié
Date de publication
09/2020
Peer-reviewed
Oui
Volume
97
Numéro
9
Pages
933-944
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
The interaction of platelet agonists with their respective membrane receptors triggers intracellular signaling, among which cytosolic ion fluxes play an important role in activation processes. While the key contribution of intercellular free calcium is accepted, sodium and potassium roles in platelet activation have been less investigated in recent studies. Here, we implemented a novel flow-cytometric method to monitor over time cytosolic free calcium, sodium, and potassium ion fluxes upon platelet activation and we demonstrate the feasibility of real-time visualization of ion kinetics, in particular with a focus on sodium and potassium. Platelets were loaded with selective ion indicators, Fluo-3 (Ca <sup>2+</sup> ), ION NaTRIUM Green-2 (Na <sup>+</sup> ), and ION Potassium Green-2 (K <sup>+</sup> ). Fluorescence was monitored by flow cytometry. After measurement of a stable baseline, platelets were activated and ion indicator fluorescence was acquired over time, up to 10 min. Platelets were activated with either thromboxane analogue U46619, ADP, thrombin, TRAP6 (PAR-1 agonist), AYPGKF (PAR-4 agonist), convulxin (collagen receptor GPVI agonist), or combinations thereof. We evaluated preanalytical parameters (in particular dye loading time and concentration) to implement an accurate method. Subsequently, we characterized cytosolic calcium, sodium, and potassium kinetics in response to platelet agonists. We observed different patterns of agonist synergism. In conclusion, the present work highlights the use of cytosolic ion monitoring by flow cytometry to investigate characteristic calcium, sodium, and potassium mobilization patterns following platelet activation. This easy technique opens a new way to analyze signaling in different platelet subpopulations and it should prove useful for investigating platelet pathophysiology. © 2020 International Society for Advancement of Cytometry.
Mots-clé
blood platelets, flow cytometry, ion transport, kinetics, platelet activation
Pubmed
Web of science
Création de la notice
02/05/2020 14:19
Dernière modification de la notice
23/11/2020 6:24
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