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Morphometric analysis of spread platelets identifies integrin αIIbβ3-specific contractile phenotype
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Morphometric analysis of spread platelets identifies integrin αIIbβ3-specific contractile phenotype
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Journal Article
Morphometric analysis of spread platelets identifies integrin αIIbβ3-specific contractile phenotype
2018
Overview
Haemostatic platelet function is intimately linked to cellular mechanics and cytoskeletal morphology. How cytoskeletal reorganizations give rise to a highly contractile phenotype that is necessary for clot contraction remains poorly understood. To elucidate this process
in vitro
, we developed a morphometric screen to quantify the spatial organization of actin fibres and vinculin adhesion sites in single spread platelets. Platelets from healthy donors predominantly adopted a bipolar morphology on fibrinogen and fibronectin, whereas distinguishable, more isotropic phenotypes on collagen type I or laminin. Specific integrin α
IIb
β
3
inhibitors induced an isotropic cytoskeletal organization in a dose-dependent manner. The same trend was observed with decreasing matrix stiffness. Circular F-actin arrangements in platelets from a patient with type II Glanzmann thrombasthenia (GT) were consistent with the residual activity of a small number of α
IIb
β
3
integrins. Cytoskeletal morphologies
in vitro
thus inform about platelet adhesion receptor identity and functionality, and integrin α
IIb
β
3
mechanotransduction fundamentally determines the adoption of a bipolar phenotype associated with contraction. Super-resolution microscopy and electron microscopies further confirmed the stress fibre-like contractile actin architecture. For the first time, our assay allows the unbiased and quantitative assessment of platelet morphologies and could help to identify defective platelet behaviour contributing to elusive bleeding phenotypes.
