Quantitation of microparticles released from coated‐platelets

Dual agonist stimulation of platelets with thrombin and convulxin results in generation of coated‐platelets, a sub‐population of cells known formerly as COAT‐platelets (collagen and thrombin). Coated‐platelets retain several procoagulant proteins on their surface and express phosphatidylserine (PS)....

Full description

Saved in:
Bibliographic Details
Published in:Journal of thrombosis and haemostasis 2005-09, Vol.3 (9), p.2081-2088
Main Authors: DALE, G. L., REMENYI, G., FRIESE, P.
Format: Article
Language:English
Subjects:
Antigens, CD - analysis
Blood Platelets - chemistry
Blood Platelets - ultrastructure
convulxin
Crotalid Venoms - pharmacology
fibrinogen
Flow Cytometry
Fluorescent Dyes
Glycocalyx - chemistry
Glycocalyx - secretion
Humans
Lectins, C-Type
Membrane Glycoproteins - analysis
Methods
microparticle
Microscopy, Fluorescence
Particle Size
Phosphatidylserines - analysis
platelet
Platelet Activation
Platelet Glycoprotein GPIb-IX Complex - analysis
Platelet Glycoprotein GPIIb-IIIa Complex - analysis
Tetraspanin-29
thrombin
Thrombin - pharmacology
thrombospondin
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Dual agonist stimulation of platelets with thrombin and convulxin results in generation of coated‐platelets, a sub‐population of cells known formerly as COAT‐platelets (collagen and thrombin). Coated‐platelets retain several procoagulant proteins on their surface and express phosphatidylserine (PS). In this report, we utilize a new methodology to demonstrate that coated‐platelets also release microparticles. Platelets were prelabeled with 2.5 μm Bodipy‐maleimide and then stimulated with convulxin plus thrombin. Microparticles, 0.3–0.5 μm in diameter, were observed by fluorescence confocal microscopy. Confocal microscopy was also used to demonstrate that microparticles were positive for glycoprotein IIb/IIIa, glycoprotein Ib, CD9, and PS, but negative for fibrinogen and thrombospondin. Furthermore, microparticles released from Bodipy‐labeled platelets were observed by flow cytometry, and activation with convulxin plus thrombin produced 15 ± 5 microparticles per coated‐platelet. In contrast, platelets stimulated with thrombin or convulxin alone produced few microparticles. Phenylarsine oxide and diamide, both of which potentiate the mitochondrial permeability transition pore and coated‐platelet production, significantly increased the number of microparticles released per coated‐platelet.
ISSN:1538-7933
1538-7836
1538-7836
DOI:10.1111/j.1538-7836.2005.01528.x