The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation

Background: Activation of the platelet-specific collagen receptor, glycoprotein (GP) VI, induces intracellular reactive oxygen species (ROS) production; however the relevance of ROS to GPVI-mediated platelet responses remains unclear. Objective: The objective of this study was to explore the role of...

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Main Authors: Walsh, T., Berndt, Michael, Carrim, N., Cowman, J., Kenny, D., Metharom, Pat
Format: Journal Article
Published: Elsevier BV 2014
Online Access:http://hdl.handle.net/20.500.11937/7108
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author Walsh, T.
Berndt, Michael
Carrim, N.
Cowman, J.
Kenny, D.
Metharom, Pat
author_facet Walsh, T.
Berndt, Michael
Carrim, N.
Cowman, J.
Kenny, D.
Metharom, Pat
author_sort Walsh, T.
building Curtin Institutional Repository
collection Online Access
description Background: Activation of the platelet-specific collagen receptor, glycoprotein (GP) VI, induces intracellular reactive oxygen species (ROS) production; however the relevance of ROS to GPVI-mediated platelet responses remains unclear. Objective: The objective of this study was to explore the role of the ROS-producing NADPH oxidase (Nox)1 and 2 complexes in GPVI-dependent platelet activation and collagen-induced thrombus formation. Methods and results: ROS production was measured by quantitating changes in the oxidation-sensitive dye, H2DCF-DA, following platelet activation with the GPVI-specific agonist, collagen related peptide (CRP). Using a pharmacological inhibitor specific for Nox1, 2-acetylphenothiazine (ML171), and Nox2 deficient mice, we show that Nox1 is the key Nox homolog regulating GPVI-dependent ROS production. Nox1, but not Nox2, was essential for CRP-dependent thromboxane (Tx)A2 production, which was mediated in part through p38 MAPK signaling; while neither Nox1 nor Nox2 was significantly involved in regulating CRP-induced platelet aggregation/integrin αIIbβ3 activation, platelet spreading, or dense granule and α-granule release (ATP release and P-selectin surface expression, respectively). Ex-vivo perfusion analysis of mouse whole blood revealed that both Nox1 and Nox2 were involved in collagen-mediated thrombus formation at arterial shear. Conclusion: Together these results demonstrate a novel role for Nox1 in regulating GPVI-induced ROS production, which is essential for optimal p38 activation and subsequent TxA2 production, providing an explanation for reduced thrombus formation following Nox1 inhibition.
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spelling curtin-20.500.11937-71082017-09-13T14:39:53Z The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation Walsh, T. Berndt, Michael Carrim, N. Cowman, J. Kenny, D. Metharom, Pat Background: Activation of the platelet-specific collagen receptor, glycoprotein (GP) VI, induces intracellular reactive oxygen species (ROS) production; however the relevance of ROS to GPVI-mediated platelet responses remains unclear. Objective: The objective of this study was to explore the role of the ROS-producing NADPH oxidase (Nox)1 and 2 complexes in GPVI-dependent platelet activation and collagen-induced thrombus formation. Methods and results: ROS production was measured by quantitating changes in the oxidation-sensitive dye, H2DCF-DA, following platelet activation with the GPVI-specific agonist, collagen related peptide (CRP). Using a pharmacological inhibitor specific for Nox1, 2-acetylphenothiazine (ML171), and Nox2 deficient mice, we show that Nox1 is the key Nox homolog regulating GPVI-dependent ROS production. Nox1, but not Nox2, was essential for CRP-dependent thromboxane (Tx)A2 production, which was mediated in part through p38 MAPK signaling; while neither Nox1 nor Nox2 was significantly involved in regulating CRP-induced platelet aggregation/integrin αIIbβ3 activation, platelet spreading, or dense granule and α-granule release (ATP release and P-selectin surface expression, respectively). Ex-vivo perfusion analysis of mouse whole blood revealed that both Nox1 and Nox2 were involved in collagen-mediated thrombus formation at arterial shear. Conclusion: Together these results demonstrate a novel role for Nox1 in regulating GPVI-induced ROS production, which is essential for optimal p38 activation and subsequent TxA2 production, providing an explanation for reduced thrombus formation following Nox1 inhibition. 2014 Journal Article http://hdl.handle.net/20.500.11937/7108 10.1016/j.redox.2013.12.023 Elsevier BV unknown
spellingShingle Walsh, T.
Berndt, Michael
Carrim, N.
Cowman, J.
Kenny, D.
Metharom, Pat
The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title_full The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title_fullStr The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title_full_unstemmed The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title_short The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation
title_sort role of nox1 and nox2 in gpvi-dependent platelet activation and thrombus formation
url http://hdl.handle.net/20.500.11937/7108