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CC BY-NC-ND 4.0 · Thromb Haemost 2017; 117(08): 1588-1600
DOI: 10.1160/TH16-12-0906
Cellular Haemostasis and Platelets
Schattauer GmbH

Differential integrin activity mediated by platelet collagen receptor engagement under flow conditions

Nicholas Pugh
1   Department of Biochemistry, University of Cambridge, Cambridge, UK
2   Department of Biomedical and Forensic Sciences, Anglia Ruskin University, Cambridge, UK
,
Ben D. Maddox
1   Department of Biochemistry, University of Cambridge, Cambridge, UK
,
Dominique Bihan
1   Department of Biochemistry, University of Cambridge, Cambridge, UK
,
Kirk A. Taylor
2   Department of Biomedical and Forensic Sciences, Anglia Ruskin University, Cambridge, UK
,
Martyn P. Mahaut-Smith
3   Department of Molecular and Cell Biology, University of Leicester, Leicester, UK
,
Richard W. Farndale
1   Department of Biochemistry, University of Cambridge, Cambridge, UK
› Author Affiliations Financial support: This work was funded by British Heart Foundation grants to NP (Project PG/14/47/30912) and RWF (Programme RG/09/003/27122).
Further Information

Publication History

Received: 05 December 2016

Accepted after major revision: 22 April 2017

Publication Date:
22 November 2017 (online)

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Summary

The platelet receptors glycoprotein (Gp)VI, integrin α2β1 and GpIb/V/IX mediate platelet adhesion and activation during thrombogenesis. Increases of intracellular Ca2+ ([Ca2+]i) are key signals during platelet activation; however, their relative importance in coupling different collagen receptors to functional responses under shear conditions remains unclear. To study shear-dependent, receptor-specific platelet responses, we used collagen or combinations of receptor-specific collagen-mimetic peptides as substrates for platelet adhesion and activation in whole human blood under arterial flow conditions and compared real-time and endpoint parameters of thrombus formation alongside [Ca2+]i measurements using confocal imaging. All three collagen receptors coupled to [Ca2+]i signals, but these varied in amplitude and temporal pattern alongside variable integrin activation. GpVI engagement produced large, sustained [Ca2+]i signals leading to realtime increases in integrins α2β1− and αIIbβ3-mediated platelet adhesion. αIIbβ3-dependent platelet aggregation was dependent on P2Y12 signalling. Co-engagement of α2β1 and GpIb/V/IX generated transient [Ca2+]i spikes and low amplitude [Ca2+]i responses that potentiated GpVI-dependent [Ca2+]i signalling. Therefore α2β1 GpIb/V/IX and GpVI synergise to generate [Ca2+]i signals that regulate platelet behaviour and thrombus formation. Antagonism of secondary signalling pathways reveals distinct, separate roles for αIIbβ3 in stable platelet adhesion and aggregation.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Platelet glycoproteins - collagen receptors - signal transduction - thrombosis - fluid shear

Author contributions

N. P. designed experiments, performed research, analysed data, and wrote the manuscript. B.M. performed research. D.B. synthesised peptides. K.T. and M.M-S wrote the manuscript. R.W.F. designed experiments, analysed data, and wrote the manuscript.


 

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