title: S1P activates store-operated calcium entry via receptor- and non-receptor-mediated pathways in vascular smooth muscle cells.
abstract: Sphingosine-1-phosphate (S1P) has been shown to modulate intracellular Ca(2+) through both G protein-coupled receptors and intracellular second messenger pathways. The precise mechanism by which S1P activates store-operated calcium entry (SOCE) in vascular smooth muscle cells (VSMCs) has not been fully characterized. Because sphingolipids and Ca(2+) modulate proliferation and constriction in VSMCs, characterizing the connection between S1P and SOCE may provide novel therapeutic targets for vascular diseases. We found that S1P triggered STIM1 puncta formation and SOCE in VSMCs. S1P-activated SOCE was inhibited by 2-aminoethoxydiphenyl borate (2-APB), diethylstilbestrol (DES), and gadolinium (Gd(3+)). SOCE was observed in VSMCs lacking either S1P(2) or S1P(3) receptors, suggesting that S1P acts via multiple signaling pathways. Indeed, both extracellular and intracellular S1P application increased the total internal reflection fluorescence signal in VSMCs cells transfected with STIM1-yellow fluorescent protein in a 2-APB-sensitive manner. These data, and the fact that 2-APB, DES, and Gd(3+) all inhibited S1P-induced cerebral artery constriction, suggest that SOCE modulates S1P-induced vasoconstriction in vivo. Finally, S1P-induced SOCE was larger in proliferative than in contractile VSMCs, correlating with increases in STIM1, Orai1, S1P(1), and S1P(3) receptor mRNA. These data demonstrate that S1P can act through both receptors and a novel intracellular pathway to activate SOCE. Because S1P-induced SOCE contributes to vessel constriction and is increased in proliferative VSMCs, it is likely that S1P/SOCE signaling in proliferative VSMCs may play a role in vascular dysfunction such as atherosclerosis and diabetes.
authors.affiliation: Stroke and Neurovascular Regulation Laboratory, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, USA.
authors.forename: Kristen Park
authors.initials: KP
authors.lastname: Hopson
authors.forename: Jessica
authors.initials: J
authors.lastname: Truelove
authors.forename: Jerold
authors.initials: J
authors.lastname: Chun
authors.forename: Yumei
authors.initials: Y
authors.lastname: Wang
authors.forename: Christian
authors.initials: C
authors.lastname: Waeber
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels; D002478:Cells, Cultured; D015870:Gene Expression; D008246:Lysophospholipids / Q000378:metabolism*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000081025:Sphingosine-1-Phosphate Receptors; D000071737:Stromal Interaction Molecule 1; D014661:Vasoconstriction / Q000502:physiology
chemical_list: D001896:Boron Compounds; D015220:Calcium Channels; D008246:Lysophospholipids; D008562:Membrane Glycoproteins; D049349:Receptors, Lysosphingolipid; D011993:Recombinant Fusion Proteins; C501285:S1pr3 protein, mouse; D000081025:Sphingosine-1-Phosphate Receptors; C100757:Stim1 protein, mouse; D000071737:Stromal Interaction Molecule 1; C533955:sphingosine-1-phosphate receptor-2, mouse; C060506:sphingosine 1-phosphate; C109986:2-aminoethoxydiphenyl borate; D013110:Sphingosine; D002118:Calcium