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Methods to study the roles of Rho GTPases in platelet function

Rivero, Francisco; Calaminus, Simon


Francisco Rivero

Simon Calaminus


Platelets are a critical cell for prevention of bleeding. Part of the response to the formation of the thrombus is the activation of the actin cytoskeleton, with an inability to effectively activate the cytoskeleton linked to thrombus formation defects and instability. The control of this process is linked to activation of the Rho GTPases, Cdc42, Rac1, and RhoA, although additional small GTPases such as Rif and Rap have been shown to play roles in platelet function.

Here we describe the methodology to accurately understand how Rho GTPases are activated in platelets. Due to the technical limitations of working with platelets, such as their lack of ability to be transfected, the majority of work has been carried out either using inhibitors of Rho GTPases or within knockout mouse models. Studies can be conducted both in suspension samples and in spread platelets. In suspension the platelets will undergo a shape change response, but will not be able to spread. In spread platelets it is possible to examine the effects of the matrix environment, such as concentration, type, and stiffness on Rho GTPase function within platelet activation and platelet spreading.

Publication Date Jul 31, 2018
Print ISSN 1064-3745
Publisher Humana Press
Peer Reviewed Peer Reviewed
Volume 1821
Pages 199-217
Book Title Methods in Molecular Biology; Rho GTPases
ISBN 9781493986118; 9781493986125
APA6 Citation Rivero, F., & Calaminus, S. (2018). Methods to study the roles of Rho GTPases in platelet function. In Methods in Molecular Biology; Rho GTPases, 199-217. Humana Press. doi:10.1007/978-1-4939-8612-5_14
Keywords Actin, RhoA, Rac1, platelets, cAMP, cGMP, pulldown assay, Y27632, Rhosin A
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