RhoA sustains integrin alpha IIbbeta 3 adhesion contacts under high shear.
The small GTPase RhoA modulates the adhesive nature of many cell types; however, despite high levels of expression in platelets, there is currently limited evidence for an important role for this small GTPase in regulating platelet adhesion processes. In this study, we have examined the role of RhoA in regulating the adhesive function of the major platelet integrin, alpha(IIb)beta(3). Our studies demonstrate that activation of RhoA occurs as a general feature of platelet activation in response to soluble agonists (thrombin, ADP, collagen), immobilized matrices (von Willebrand factor (vWf), fibrinogen) and high shear stress. Blocking the ligand binding function of integrin alpha(IIb)beta(3), by pretreating platelets with c7E3 Fab, demonstrated the existence of integrin alpha(IIb)beta(3)-dependent and -independent mechanisms regulating RhoA activation. Inhibition of RhoA (C3 exoenzyme) or its downstream effector Rho kinase had no effect on integrin alpha(IIb)beta(3) activation induced by soluble agonists or adhesive substrates, however, both inhibitors reduced shear-dependent platelet adhesion on immobilized vWf and shear-induced platelet aggregation in suspension. Detailed analysis of the sequential adhesive steps required for stable platelet adhesion on a vWf matrix under shear conditions revealed that RhoA did not regulate platelet tethering to vWf or the initial formation of integrin alpha(IIb)beta(3) adhesion contacts but played a major role in sustaining stable platelet-matrix interactions. These studies define a critical role for RhoA in regulating the stability of integrin alpha(IIb)beta(3) adhesion contacts under conditions of high shear stress.
Schoenwaelder
Simone M
SM
Department of Medicine, Australian Centre for Blood Diseases, Monash University, Box Hill Hospital, Arnold St., Box Hill, Victoria 3128, Australia. Simone.Schoenwaelder@med.monash.edu.au
Hughan
Sascha C
SC
Boniface
Karen
K
Fernando
Sujanie
S
Holdsworth
Melissa
M
Thompson
Philip E
PE
Salem
Hatem H
HH
Jackson
Shaun P
SP
D001792:Blood Platelets / Q000378:metabolism*; D002448:Cell Adhesion; D002478:Cells, Cultured; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000502:physiology*; D014841:von Willebrand Factor / Q000378:metabolism
D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D014841:von Willebrand Factor; D020742:rhoA GTP-Binding Protein