Journal
JOURNAL OF THROMBOSIS AND HAEMOSTASIS
Volume 3, Issue 9, Pages 2081-2088Publisher
WILEY
DOI: 10.1111/j.1538-7836.2005.01528.x
Keywords
convulxin; fibrinogen; microparticle; platelet; thrombin; thrombospondin
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Funding
- NHLBI NIH HHS [HL68129] Funding Source: Medline
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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 mu m Bodipy-maleimide and then stimulated with convulxin plus thrombin. Microparticles, 0.3-0.5 mu m in diameter, were observed by fluorescence confocal microscopy. Confocal microscopy was also used to demonstrate that microparticles were positive for glycoprotein Ilb/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.
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