Identification of a novel, actin-rich structure, the actin nodule, in the early stages of platelet spreading

Simon D.J. Calaminus, S. Thomas, O. J.T. McCarty, L. M. Machesky, S. P. Watson

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

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 β1- and β3- 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.

Original languageEnglish (US)
Pages (from-to)1944-1952
Number of pages9
JournalJournal of Thrombosis and Haemostasis
Volume6
Issue number11
DOIs
StatePublished - Oct 24 2008

Keywords

  • Actin cytoskeleton
  • Arp2/3
  • Platelet spreading
  • Rho kinase
  • Src kinases

ASJC Scopus subject areas

  • Hematology

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