Modulation of endothelial cell migration by extracellular nucleotides

Elzbieta Kaczmarek; Laurie Erb; Katarzyna Koziak; Robert Jarzyna; Marcia R. Wink; Olaf Guckelberger; Krzysztof J. Blusztajn; Vickery Trinkaus-Randall; Gary A. Weisman; Simon C. Robson

doi:10.1160/TH04-09-0576

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 93(04): 735-742
DOI: 10.1160/TH04-09-0576

Endothelium and Vascular Development

Schattauer GmbH

Modulation of endothelial cell migration by extracellular nucleotides

Involvement of focal adhesion kinase and phosphatidylinositol 3-kinase-mediated pathways

Elzbieta Kaczmarek

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Laurie Erb

²University of Missouri-Columbia, Columbia, Missouri, USA

,

Katarzyna Koziak

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

³Warsaw Medical University, Warsaw, Poland

,

Robert Jarzyna

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Marcia R. Wink

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Olaf Guckelberger

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Krzysztof J. Blusztajn

⁴Boston University School of Medicine, Boston, Massachusetts, USA

,

Vickery Trinkaus-Randall

⁴Boston University School of Medicine, Boston, Massachusetts, USA

,

Gary A. Weisman

²University of Missouri-Columbia, Columbia, Missouri, USA

,

Simon C. Robson

¹Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

› Author Affiliations

Abstract

Summary

Extracellular nucleotides bind to type-2 purinergic/pyrimidinergic (P2) receptors that mediate various responses, such as cell activation, proliferation and apoptosis, implicated in inflammatory processes. The role of P2 receptors and their associated signal transduction pathways in endothelial cell responses has not been fully investigated. Here, it is shown that stimulation of human umbilical vein endothelial cells (HUVEC) with extracellular ATP or UTP increased intracellular free calcium ion concentrations ([Ca²⁺]_i), induced phosphorylation of focal adhesion kinase (FAK), p130^cas and paxillin, and caused cytoskeletal rearrangements with consequent cell migration. Furthermore, UTP increased migration of HUVEC in a phosphatidylinositol 3-kinase (PI3-K)-dependent manner. BAPTA or thapsigargin inhibited the extracellular nucleotide-induced increase in [Ca²⁺]_i, a response crucial for both FAK phosphorylation and cell migration. Furthermore, long-term exposure of HUVEC to ATP and UTP, agonists of the G protein-coupled P2Y2 and P2Y4 receptor subtypes, caused upregulation of α_v integrin expression, a cell adhesion molecule known to directly interact with P2Y2 receptors. Our results suggest that extracellular nucleotides modulate signaling pathways in HUVEC influencing cell functions, such as cytoskeletal changes, cellular adhesion and motility, typically associated with integrin-activation and the action of growth factors. We propose that P2Y2 and possibly P2Y4 receptors mediate those responses that are important in vascular inflammation, atherosclerosis and angiogenesis.

Keywords

Endothelial cells - extracellular nucleotides - migration - P2 receptors - FAK

Full Text

References

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