title: Sphingosine-1-phosphate receptor 3 mediates sphingosine-1-phosphate induced release of weibel-palade bodies from endothelial cells.
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.
authors.affiliation: Department of Plasma Proteins, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Kathinka W E M
authors.initials: KW
authors.lastname: van Hooren
authors.affiliation: Department of Pharmacology and Pharmacotherapy, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Léon J A
authors.initials: LJ
authors.lastname: Spijkers
authors.affiliation: Department of Plasma Proteins, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Dorothee
authors.initials: D
authors.lastname: van Breevoort
authors.affiliation: Department of Molecular Cell Biology, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Mar
authors.initials: M
authors.lastname: Fernandez-Borja
authors.affiliation: Department of Plasma Proteins, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Ruben
authors.initials: R
authors.lastname: Bierings
authors.affiliation: Department of Molecular Cell Biology, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Jaap D
authors.initials: JD
authors.lastname: van Buul
authors.affiliation: Department of Pharmacology and Pharmacotherapy, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Astrid E
authors.initials: AE
authors.lastname: Alewijnse
authors.affiliation: Department of Pharmacology and Pharmacotherapy, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Stephan L M
authors.initials: SL
authors.lastname: Peters
authors.affiliation: Department of Plasma Proteins, Sanquin-AMC Landsteiner Laboratory, Amsterdam, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Voorberg
mesh_terms: D000577:Amides / Q000494:pharmacology; D002460:Cell Line; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism*; D000081025:Sphingosine-1-Phosphate Receptors; D020302:Weibel-Palade Bodies / Q000378:metabolism*
chemical_list: D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; D049349:Receptors, Lysosphingolipid; C477362:S1PR1 protein, human; D000081025:Sphingosine-1-Phosphate Receptors; C577483:sphingosine-1-phosphate receptor-3, human; C108830:Y 27632