Delayed-onset of procoagulant signalling revealed by kinetic analysis of COAT platelet formation.

Détails

ID Serval
serval:BIB_FB7ADCDA152B
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Delayed-onset of procoagulant signalling revealed by kinetic analysis of COAT platelet formation.
Périodique
Thrombosis and haemostasis
Auteur(s)
Alberio L., Ravanat C., Hechler B., Mangin P.H., Lanza F., Gachet C.
ISSN
2567-689X (Electronic)
ISSN-L
0340-6245
Statut éditorial
Publié
Date de publication
02/06/2017
Peer-reviewed
Oui
Volume
117
Numéro
6
Pages
1101-1114
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
The combined action of collagen and thrombin induces the formation of COAT platelets, which are characterised by a coat of procoagulant and adhesive molecules on their surface. Although recent work has started to highlight their clinical relevance, the exact mechanisms regulating the formation of procoagulant COAT platelets remain unclear. Therefore, we employed flow cytometry in order to visualise in real time surface and intracellular events following simultaneous platelet activation with convulxin and thrombin. After a rapid initial response pattern characterised by the homogenous activation of the fibrinogen receptor glycoprotein IIb/IIIa in all platelets, starting with a delay of about 2 minutes an increasing fraction transforms to procoagulant COAT platelets. Their surface is characterised by progressive loss of PAC-1 binding, expression of negative phospholipids and retention of α-granule von Willebrand factor. Intracellular events in procoagulant COAT platelets are a marked increase of free calcium into the low micromolar range, concomitantly with early depolarisation of the mitochondrial membrane and activation of caspase-3, while non-COAT platelets keep the intracellular free calcium in the nanomolar range and maintain an intact mitochondrial membrane. We show for the first time that the flow-cytometrically distinct fractions of COAT and non-COAT platelets differentially phosphorylate two signalling proteins, PKCα and p38MAPK, which may be involved in the regulation of the different calcium fluxes observed in COAT versus non-COAT platelets. This study demonstrates the utility of concomitant cellular and signalling evaluation using flow cytometry in order to further dissect the mechanisms underlying the dichotomous platelet response observed after collagen/thrombin stimulation.
Mots-clé
Blood Coagulation, Blood Platelets/metabolism, Calcium/metabolism, Cell Separation, Cells, Cultured, Collagen/metabolism, Crotalid Venoms/metabolism, Dual Specificity Phosphatase 2/metabolism, Flow Cytometry, Humans, Lectins, C-Type/metabolism, Mitochondria/metabolism, Platelet Activation, Platelet Glycoprotein GPIIb-IIIa Complex/metabolism, Protein Binding, Protein Kinase C/metabolism, Signal Transduction, Thrombin/metabolism, p38 Mitogen-Activated Protein Kinases/metabolism, von Willebrand Factor/metabolism, COAT, Platelets, apoptosis, calcium, flow cytometry, intracellular signalling, phosphorylation, procoagulant, surface
Pubmed
Web of science
Création de la notice
25/04/2017 17:25
Dernière modification de la notice
20/08/2019 16:26
Données d'usage