Markers of platelet apoptosis: methodology and applications

Apoptosis, or programmed cell death, is a physiological mechanism that serves for controlled deletion of damaged cells. While long attributed exclusively to nucleated cells, over recent years it has been recognized that apoptosis also occurs in anucleate platelets. We describe here experiences of de...

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Bibliographic Details
Published in:Journal of thrombosis and thrombolysis 2012-05, Vol.33 (4), p.397-411
Main Authors: Gyulkhandanyan, Armen V., Mutlu, Asuman, Freedman, John, Leytin, Valery
Format: Article
Language:English
Subjects:
Apoptosis - physiology
Biomarkers - metabolism
Blood Platelets - metabolism
Blood Platelets - ultrastructure
Blotting, Western - methods
Cardiology
Electron Microscope Tomography - methods
Flow Cytometry - methods
Hematology
Humans
Inhibitor of Apoptosis Proteins - metabolism
Medicine
Medicine & Public Health
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Summary:Apoptosis, or programmed cell death, is a physiological mechanism that serves for controlled deletion of damaged cells. While long attributed exclusively to nucleated cells, over recent years it has been recognized that apoptosis also occurs in anucleate platelets. We describe here experiences of determining markers of apoptosis in human platelets treated in vitro with pro-apoptotic chemical and physical stimuli. These include depolarization of mitochondrial inner membrane, cytochrome c release, expression of pro-apoptotic and anti-apoptotic proteins of Bcl-2 family, activation of apoptosis executioner caspase-3, exposure of phosphatidylserine, platelet shrinkage, fragmentation to microparticles, blebbing and filopod extrusion on the platelet surface. These assays serve to characterize platelet apoptosis in different cellular compartments (mitochondria, cytosol and plasma membrane) and at the whole-cell level. Methods commonly employed in studies of platelet apoptosis markers include flow cytometry, Western blot analysis and electron microscopy. An integrated methodological approach, based on determination of different platelet apoptosis markers, represents a useful tool for examining platelet apoptosis in various physiological and pathological settings.
ISSN:0929-5305
1573-742X
DOI:10.1007/s11239-012-0688-8