Sphingosine-1-phosphate receptor 3 mediates sphingosine-1-phosphate induced release of weibel-palade bodies from endothelial cells

PLoS One. 2014 Mar 14;9(3):e91346. doi: 10.1371/journal.pone.0091346. eCollection 2014.

Abstract

Sphingosine-1-phosphate (S1P) is an agonist for five distinct G-protein coupled receptors, that is released by platelets, mast cells, erythrocytes and endothelial cells. S1P promotes endothelial cell barrier function and induces release of endothelial cell-specific storage-organelles designated Weibel-Palade bodies (WPBs). S1P-mediated enhancement of endothelial cell barrier function is dependent on S1P receptor 1 (S1PR1) mediated signaling events that result in the activation of the small GTPase Rac1. Recently, we have reported that Rac1 regulates epinephrine-induced WPB exocytosis following its activation by phosphatidylinositol-3,4,5-triphosphate-dependent Rac exchange factor 1 (PREX1). S1P has also been described to induce WPB exocytosis. Here, we confirm that S1P induces release of WPBs using von Willebrand factor (VWF) as a marker. Using siRNA mediated knockdown of gene expression we show that S1PR1 is not involved in S1P-mediated release of WPBs. In contrast depletion of the S1PR3 greatly reduced S1P-induced release of VWF. S1P-mediated enhancement of endothelial barrier function was not affected by S1PR3-depletion whereas it was greatly impaired in cells lacking S1PR1. The Rho kinase inhibitor Y27632 completely abrogated S1P-mediated release of VWF. Also, the calcium chelator BAPTA-AM significantly reduced S1P-induced release of VWF. Our findings indicate that S1P-induced release of haemostatic, inflammatory and angiogenic components stored within WPBs depends on the S1PR3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / pharmacology
  • Cell Line
  • Down-Regulation
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Gene Expression
  • Humans
  • Protein Binding
  • Pyridines / pharmacology
  • Receptors, Lysosphingolipid / genetics
  • Receptors, Lysosphingolipid / metabolism*
  • Sphingosine-1-Phosphate Receptors
  • Weibel-Palade Bodies / metabolism*

Substances

  • Amides
  • Enzyme Inhibitors
  • Pyridines
  • Receptors, Lysosphingolipid
  • S1PR1 protein, human
  • Sphingosine-1-Phosphate Receptors
  • sphingosine-1-phosphate receptor-3, human
  • Y 27632

Grant support

This study was supported by grants from the Landsteiner Foundation for Blood Transfusion Research (LSBR 08.19, LSBR 12.44). RB was supported by a European Hematology Association Research Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.