Journal
JOURNAL OF THROMBOSIS AND HAEMOSTASIS
Volume 6, Issue 11, Pages 1944-1952Publisher
WILEY-BLACKWELL
DOI: 10.1111/j.1538-7836.2008.03141.x
Keywords
actin cytoskeleton; Arp2/3; platelet spreading; Rho kinase; Src kinases
Categories
Funding
- British Heart Foundation
- Wellcome Trust
- MRC
- American Heart Association
- BHF
- AHA [0665512Z]
- Medical Research Council [G117/569] Funding Source: researchfish
- MRC [G117/569] Funding Source: UKRI
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Background: During platelet spreading, the actin cytoskeleton undergoes marked changes, forming filopodia, lamellipodia and stress fibres. In the present study, we report the identification of a novel actin-rich structure, termed an actin nodule, which appears prior to lamellipodia and stress fibre formation. Methods: Platelet spreading was monitored using human platelets and mouse GFP-actin platelets using real-time and end-point DIC, and fluorescent and electron microscopy (EM). Results: We identified a small, novel actin structure, the actin nodule, in the early stages of adhesion and spreading, which we hypothesize to be a precursor of lamellipodia and stress fibres. Nodule formation shows an inverse correlation to Rho kinase and myosin-II activity, is independent of PI3-kinase, but dependent on Src kinase activity. Actin nodules contain multiple proteins, including Arp2/3, Fyn, Rac, and beta 1- and beta 3- integrins, but not Src. EM analysis revealed that actin filaments extend in all directions from the nodules. Actin nodules are present on multiple matrices, including fibrinogen, laminin and VWF + botrocetin. Conclusion: This work identifies a novel platelet actin structure, which we propose is a precursor to both lamellipodia and stress fibres and acts to drive platelet spreading.
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