Synergistic effect of signaling from receptors of soluble platelet agonists and outside-in signaling in formation of a stable fibrinogen-integrin αiIbβ3-actin cytoskeleton complex

Ivan Budnik, Boris Shenkman, Naphtali Savion

Research output: Contribution to journalArticlepeer-review

Abstract

Introduction Thrombus formation in the injured vessel wall is a highly complex process involving various blood-born components that go through specific temporal and spatial changes as observed by intravital videomicroscopy. Platelets bind transiently to the developing thrombus and may either become stably incorporated into or disengage from the thrombus. The aim of the present study was to reveal the processes involved in the formation of a stable thrombus. Methods Platelet-rich plasma and washed platelets were studied by the aggregometer. The aggregate stability was challenged by eptifibatide. Platelet Triton-insoluble fraction was prepared and the actin and αIIb content in the cytoskeleton was analyzed by western blot. Results Maximal actin polymerization is achieved 1 min after platelet activation while maximal αIIbβ3-actin cytoskeleton association requires 5 to 10 min of activation and fibrinogen-mediated platelet-to-platelet bridging. Thus, actin polymerization is dependent on platelet activation and requires neither αIIbβ3 integrin occupation nor platelet aggregation. Formation of a stable aggregate requires platelet activation for more than 1 min, complete increase in actin cytoskeleton fraction and partial association of αIIbβ3 with the actin cytoskeleton. However, direct αIIbβ3 activation is not sufficient for cytoskeleton complex formation. Thus, stable αIIbβ3-fibrinogen interaction, representing stable aggregate, is achieved after more than 1 min agonist activation, involving inside-out and outside-in signaling but not after direct integrin activation, involving only outside-in signaling. Conclusions Formation of a stable fibrinogen-αIIbβ3-actin cytoskeleton complex is the result of the combined effect of platelet stimulation by soluble agonists, activation of αIIbβ3, fibrinogen binding and platelet-to-platelet bridging.

Original languageEnglish
Pages (from-to)114-120
Number of pages7
JournalThrombosis Research
Volume135
Issue number1
DOIs
StatePublished - 1 Jan 2015

Keywords

  • actin cytoskeleton
  • fibrinogen-integrin αIIbβ3 binding
  • platelet aggregation

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