Actin polymerisation regulates thrombin-evoked Ca(2+) signalling after activation of PAR-4 but not PAR-1 in human platelets

Matthew T Harper, Stewart O Sage

Research output: Contribution to journalArticle (Academic Journal)peer-review

13 Citations (Scopus)

Abstract

The role of actin polymerisation in regulating thrombin-evoked Ca(2+) signalling was investigated in human platelets. We have previously reported that cytochalasin D (Cyt D) inhibits thapsigargin-evoked store-operated Ca(2+) entry (SOCE), which is believed to contribute a major component of thrombin-evoked Ca(2+) entry in platelets. In contrast, Cyt D increased thrombin-evoked Ca(2+) entry to 147.5 +/- 9.2% and Sr(2+) entry to 134.2 +/- 6.4% of control. Similar results were obtained with latrunculin A. This potentiation was not affected if protein kinase C was inhibited using Ro-31-8220, suggesting that it did not involve PKC-dependent non-capacitative Ca(2+) entry. Ca(2+) entry evoked by the PAR-4 agonist, AYPGKF, was increased to 133.7 +/- 12.8% of control by Cyt D, whereas Ca(2+) signalling evoked by the PAR-1 agonist, SFLLRN, was unaffected. The PAR-4 antagonist, tcY-NH(2), abolished the effect of Cyt D on thrombin-evoked Ca(2+) entry. Biotinylation of cell-surface proteins showed that PAR-4 was internalised after stimulation by thrombin. Cyt D reduced this internalisation. These data suggest that Cyt D prevents the internalisation of PAR-4, which may lead to prolonged signalling from this receptor. This may mask a direct effect of Cyt D on the activation of SOCE after the activation of PAR-4.
Original languageEnglish
Pages (from-to)134-42
Number of pages9
JournalPlatelets
Volume17
Issue number3
DOIs
Publication statusPublished - May 2006

Keywords

  • Thrombin
  • Calcium
  • Thiazoles
  • Humans
  • Receptors, Proteinase-Activated
  • Cytochalasin D
  • Bicyclo Compounds, Heterocyclic
  • Biotinylation
  • Cytoskeleton
  • Indoles
  • Blood Platelets
  • Peptide Fragments
  • Actins
  • Fura-2
  • Strontium
  • Protein Kinase C
  • Oligopeptides
  • Thiazolidines
  • Receptor, PAR-1
  • Calcium Signaling
  • Receptors, Thrombin

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