Stimulation of intracellular sphingosine-1-phosphate production by G-protein-coupled sphingosine-1-phosphate receptors

Eur J Pharmacol. 2001 Mar 2;414(2-3):145-54. doi: 10.1016/s0014-2999(01)00789-0.


Recently, a family of G-protein-coupled receptors named endothelial differentiation gene (Edg) receptor family has been identified, which are specifically activated by the two serum lipids, sphingosine-1-phosphate and lysophosphatidic acid. Sphingosine-1-phosphate can also act intracellularly to release Ca2+ from intracellular stores. Since in several cell types, G-protein-coupled lysophosphatidic acid or sphingosine-1-phosphate receptors mobilize Ca2+ in the absence of a measurable phospholipase C stimulation, it was analysed here whether intracellular sphingosine-1-phosphate production was the signalling mechanism used by extracellular sphingosine-1-phosphate for mobilization of stored Ca2+. Sphingosine-1-phosphate and the low affinity sphingosine-1-phosphate receptor agonist, sphingosylphosphorylcholine, induced a rapid, transient and nearly complete pertussis toxin-sensitive Ca2+ mobilization in human embryonic kidney (HEK-293) cells. The G-protein-coupled sphingosine-1-phosphate receptors, Edg-1, Edg-3 and Edg-5, were found to be endogenously expressed in these cells. Most interestingly, sphingosine-1-phosphate and sphingosylphosphorylcholine did not induce a measurable production of inositol-1,4,5-trisphosphate or accumulation of inositol phosphates. Instead, sphingosine-1-phosphate and sphingosylphosphorylcholine induced a rapid and transient increase in production of intracellular sphingosine-1-phosphate with a maximum of about 1.4-fold at 30 s. Stimulation of sphingosine-1-phosphate formation by sphingosine-1-phosphate and sphingosylphosphorylcholine was fully blocked by pertussis toxin, indicating that extracellular sphingosine-1-phosphate via endogenously expressed G(i)-coupled receptors induces a stimulation of intracellular sphingosine-1-phosphate production. As sphingosine-1-phosphate- and sphingosylphosphorylcholine-induced increases in intracellular Ca2+ were blunted by sphingosine kinase inhibitors, this sphingosine-1-phosphate production appears to mediate Ca2+ signalling by extracellular sphingosine-1-phosphate and sphingosylphosphorylcholine in HEK-293 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium Signaling / drug effects
  • Calcium Signaling / physiology*
  • Carbachol / pharmacology
  • Cell Line
  • Cholinergic Agonists / pharmacology
  • DNA-Binding Proteins / metabolism*
  • GTP-Binding Proteins / drug effects
  • GTP-Binding Proteins / metabolism
  • Humans
  • I-kappa B Proteins*
  • Immediate-Early Proteins / metabolism*
  • Lysophospholipids*
  • NF-KappaB Inhibitor alpha
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / pharmacology
  • Phosphotransferases (Alcohol Group Acceptor) / drug effects
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Receptors, Cell Surface / drug effects
  • Receptors, Cell Surface / metabolism*
  • Receptors, G-Protein-Coupled*
  • Receptors, Lysophospholipid
  • Sphingosine / analogs & derivatives*
  • Sphingosine / metabolism*
  • Sphingosine / pharmacology
  • Type C Phospholipases / drug effects
  • Type C Phospholipases / metabolism


  • Cholinergic Agonists
  • DNA-Binding Proteins
  • I-kappa B Proteins
  • Immediate-Early Proteins
  • Lysophospholipids
  • NFKBIA protein, human
  • Receptors, Cell Surface
  • Receptors, G-Protein-Coupled
  • Receptors, Lysophospholipid
  • sphingosine phosphorylcholine
  • Phosphorylcholine
  • NF-KappaB Inhibitor alpha
  • sphingosine 1-phosphate
  • Carbachol
  • Phosphotransferases (Alcohol Group Acceptor)
  • sphingosine kinase
  • Type C Phospholipases
  • GTP-Binding Proteins
  • Sphingosine