P2X1-mediated activation of extracellular signal-regulated kinase 2 contributes to platelet secretion and aggregation induced by collagen

Cécile Oury, E. Tóth-Zsámboki, Jos Vermylen, Marc F. Hoylaerts

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

Adenosine triphosphate (ATP) and its stable analog, α,β-methylene ATP, activate the platelet P2X1 ion channel, causing a rapid Ca++ influx. Here, we show that, in washed apyrase-treated platelets, α,β-methylene ATP elicits reversible extracellular signal-regulated kinase 2 (ERK2) phosphorylation through a Ca++- and protein kinase C-dependent pathway. In contrast, high-performance liquid chromatography-purified adenosine diphosphate (ADP) did not trigger ERK2 phosphorylation. α,β-Methylene ATP also activated the ERK2 pathway in P2X1-transfected HEK293 cells but not in cells expressing mutated P2X1delL nonfunctional channels. Because ATP released from the dense granules during platelet activation contributes to platelet aggregation elicited by low doses of collagen, and because collagen causes ERK2 phosphorylation, we have investigated the role of P2X1-mediated ERK2 activation in these platelet responses. We found that the antagonism of P2X1 with ADP or desensitization of this ion channel with α,β-methylene ATP both resulted in impaired ERK2 phosphorylation, ATP secretion, and platelet aggregation induced by low concentrations of collagen (≤1 μg/mL) without affecting the minor early dense granule release. Selective MEK1/2 inhibition by U-0126 and Ca++ chelation with EGTA (ethyleneglycoltetraacetic acid) behaved similarly, whereas the PKC inhibitor GF109203-X totally prevented collagen-induced secretion and ERK2 activation. In contrast, when elicited by high collagen concentrations (2 μg/mL), platelet aggregation and secretion no longer depended on P2X1 or ERK2 activation, as shown by the lack of their inhibition by α,β-methylene ATP or U-0126. We thus conclude that mild platelet stimulation with collagen rapidly releases ATP, which activates the P2X1-PKC-ERK2 pathway. This process enhances further degranulation of the collagen-primed granules allowing platelet aggregation to be completed.

Original languageEnglish
Pages (from-to)2499-2505
Number of pages7
JournalBlood
Volume100
Issue number7
DOIs
Publication statusPublished - Oct 1 2002

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Mitogen-Activated Protein Kinase 1
Platelets
Platelet Aggregation
Collagen
Agglomeration
Adenosine Triphosphate
Chemical activation
Phosphorylation
Blood Platelets
Platelet Activation
Ion Channels
Adenosine Diphosphate
Apyrase
HEK293 Cells
High performance liquid chromatography
Chelation
Protein Kinase C
High Pressure Liquid Chromatography
Cells
Acids

ASJC Scopus subject areas

  • Hematology

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P2X1-mediated activation of extracellular signal-regulated kinase 2 contributes to platelet secretion and aggregation induced by collagen. / Oury, Cécile; Tóth-Zsámboki, E.; Vermylen, Jos; Hoylaerts, Marc F.

In: Blood, Vol. 100, No. 7, 01.10.2002, p. 2499-2505.

Research output: Contribution to journalArticle

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