Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation.
Journal: 2000/November - Journal of Clinical Investigation
ISSN: 0021-9738
Abstract:
Sphingolipid signaling pathways have been implicated in many critical cellular events. Sphingosine-1-phosphate (SPP), a sphingolipid metabolite found in high concentrations in platelets and blood, stimulates members of the endothelial differentiation gene (Edg) family of G protein-coupled receptors and triggers diverse effects, including cell growth, survival, migration, and morphogenesis. To determine the in vivo functions of the SPP/Edg signaling pathway, we disrupted the Edg1 gene in mice. Edg1(-/-) mice exhibited embryonic hemorrhage leading to intrauterine death between E12.5 and E14.5. Vasculogenesis and angiogenesis appeared normal in the mutant embryos. However, vascular maturation was incomplete due to a deficiency of vascular smooth muscle cells/pericytes. We also show that Edg-1 mediates an SPP-induced migration response that is defective in mutant cells due to an inability to activate the small GTPase, Rac. Our data reveal Edg-1 to be the first G protein-coupled receptor required for blood vessel formation and show that sphingolipid signaling is essential during mammalian development.
Relations:
Content
Citations
(320)
References
(60)
Chemicals
(7)
Genes
(15)
Organisms
(3)
Processes
(4)
Anatomy
(5)
Affiliates
(1)
Similar articles
Articles by the same authors
Discussion board
J Clin Invest 106(8): 951-961

Edg-1, the G protein–coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation

+5 authors
Genetics of Development and Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, Maryland, USADepartment of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, DC, USACenter for Vascular Biology, Department of Physiology, University of Connecticut Health Center, Farmington, Connecticut, USA
Address correspondence to: Richard L. Proia, Building 10, Room 9N-314, National Institutes of Health, 10 Center DR MSC 1821, Bethesda, Maryland 20892-1821, USA. Phone: (301) 496-4391; Fax: (301) 496-9878; E-mail: vog.hin@aiorp.
Address correspondence to: Richard L. Proia, Building 10, Room 9N-314, National Institutes of Health, 10 Center DR MSC 1821, Bethesda, Maryland 20892-1821, USA. Phone: (301) 496-4391; Fax: (301) 496-9878; E-mail: vog.hin@aiorp.
Received 2000 Jul 28; Accepted 2000 Sep 12.

Abstract

Sphingolipid signaling pathways have been implicated in many critical cellular events. Sphingosine-1-phosphate (SPP), a sphingolipid metabolite found in high concentrations in platelets and blood, stimulates members of the endothelial differentiation gene (Edg) family of G protein–coupled receptors and triggers diverse effects, including cell growth, survival, migration, and morphogenesis. To determine the in vivo functions of the SPP/Edg signaling pathway, we disrupted the Edg1 gene in mice. Edg1–/– mice exhibited embryonic hemorrhage leading to intrauterine death between E12.5 and E14.5. Vasculogenesis and angiogenesis appeared normal in the mutant embryos. However, vascular maturation was incomplete due to a deficiency of vascular smooth muscle cells/pericytes. We also show that Edg-1 mediates an SPP-induced migration response that is defective in mutant cells due to an inability to activate the small GTPase, Rac. Our data reveal Edg-1 to be the first G protein–coupled receptor required for blood vessel formation and show that sphingolipid signaling is essential during mammalian development.

Abstract

Acknowledgments

We thank J. Van Brocklyn for help in the early stages of this project and A. Howard for producing the figures.

Acknowledgments

References

  • 1. Shayman JAPerspectives in basic science: sphingolipids. Kidney Int. 2000;58:11–26.[PubMed][Google Scholar]
  • 2. Huwiler A, Kolter T, Pfeilschifter J, Sandhoff KPhysiology and pathophysiology of sphingolipid metabolism and signaling. Biochim Biophys Acta. 2000;1485:63–99.[PubMed][Google Scholar]
  • 3. Spiegel S, Merrill AH., Jr Sphingolipid metabolism and cell growth regulation. FASEB J. 1996;10:1388–1397.[PubMed]
  • 4. Lee MJ, et al Sphingosine-1-phosphate as a ligand for the G protein-coupled receptor EDG-1. Science. 1998;279:1552–1555.[PubMed][Google Scholar]
  • 5. Hla T, Maciag TAn abundant transcript induced in differentiating human endothelial cells encodes a polypeptide with structural similarities to G-protein–coupled receptors. J Biol Chem. 1990;265:9308–9313.[PubMed][Google Scholar]
  • 6. Goetzl EJ, An SDiversity of cellular receptors and functions for the lysophospholipid growth factors lysophosphatidic acid and sphingosine 1-phosphate. FASEB J. 1998;12:1589–1598.[PubMed][Google Scholar]
  • 7. Spiegel S, Milstien SFunctions of a new family of sphingosine-1-phosphate receptors. Biochim Biophys Acta. 2000;1484:107–116.[PubMed][Google Scholar]
  • 8. Hla T, et al Sphingosine-1-phosphate: extracellular mediator or intracellular second messenger? Biochem Pharmacol. 1999;58:201–207.[PubMed][Google Scholar]
  • 9. Chun J, Contos JJ, Munroe DA growing family of receptor genes for lysophosphatidic acid (LPA) and other lysophospholipids (LPs) Cell Biochem Biophys. 1999;30:213–242.[PubMed][Google Scholar]
  • 10. Moolenaar WHBioactive lysophospholipids and their G protein–coupled receptors. Exp Cell Res. 1999;253:230–238.[PubMed][Google Scholar]
  • 11. Lee MJ, Evans M, Hla TThe inducible G protein–coupled receptor edg-1 signals via the G(i)/mitogen-activated protein kinase pathway. J Biol Chem. 1996;271:11272–11279.[PubMed][Google Scholar]
  • 12. Ancellin N, Hla TDifferential pharmacological properties and signal transduction of the sphingosine 1-phosphate receptors EDG-1, EDG-3, and EDG-5. J Biol Chem. 1999;274:18997–19002.[PubMed][Google Scholar]
  • 13. An S, Bleu T, Zheng YTransduction of intracellular calcium signals through G protein–mediated activation of phospholipase C by recombinant sphingosine 1-phosphate receptors. Mol Pharmacol. 1999;55:787–794.[PubMed][Google Scholar]
  • 14. Windh RT, et al Differential coupling of the sphingosine 1-phosphate receptors Edg-1, Edg-3, and H218/Edg-5 to the g(i), g(q), and G(12) families of heterotrimeric G proteins. J Biol Chem. 1999;274:27351–27358.[PubMed][Google Scholar]
  • 15. Hecht JH, Weiner JA, Post SR, Chun JVentricular zone gene-1 (vzg-1) encodes a lysophosphatidic acid receptor expressed in neurogenic regions of the developing cerebral cortex. J Cell Biol. 1996;135:1071–1083.[Google Scholar]
  • 16. Liu CH, Hla TThe mouse gene for the inducible G-protein-coupled receptor edg-1. Genomics. 1997;43:15–24.[PubMed][Google Scholar]
  • 17. Graler MH, Bernhardt G, Lipp MEDG6, a novel G-protein–coupled receptor related to receptors for bioactive lysophospholipids, is specifically expressed in lymphoid tissue. Genomics. 1998;53:164–169.[PubMed][Google Scholar]
  • 18. Zhang G, Contos JJ, Weiner JA, Fukushima N, Chun JComparative analysis of three murine G-protein coupled receptors activated by sphingosine-1-phosphate. Gene. 1999;227:89–99.[PubMed][Google Scholar]
  • 19. Nehls M, et al Two genetically separable steps in the differentiation of thymic epithelium. Science. 1996;272:886–889.[PubMed][Google Scholar]
  • 20. Mountford PS, Smith AGInternal ribosome entry sites and dicistronic RNAs in mammalian transgenesis. Trends Genet. 1995;11:179–184.[PubMed][Google Scholar]
  • 21. Liu Y, et al A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder. J Clin Invest. 1999;103:497–505.[Google Scholar]
  • 22. Wang F, Nohara K, Olivera A, Thompson EW, Spiegel SInvolvement of focal adhesion kinase in inhibition of motility of human breast cancer cells by sphingosine 1-phosphate. Exp Cell Res. 1999;247:17–28.[PubMed][Google Scholar]
  • 23. Benard V, Bohl BP, Bokoch GMCharacterization of rac and cdc42 activation in chemoattractant-stimulated human neutrophils using a novel assay for active GTPases. J Biol Chem. 1999;274:13198–13204.[PubMed][Google Scholar]
  • 24. Lee MJ, et al Vascular endothelial cell adherens junction assembly and morphogenesis induced by sphingosine-1-phosphate. Cell. 1999;99:301–312.[PubMed][Google Scholar]
  • 25. Carmeliet P, et al Targeted deficiency or cytosolic truncation of the VE-cadherin gene in mice impairs VEGF-mediated endothelial survival and angiogenesis. Cell. 1999;98:147–157.[PubMed][Google Scholar]
  • 26. Carmeliet PMechanisms of angiogenesis and arteriogenesis. Nat Med. 2000;6:389–395.[PubMed][Google Scholar]
  • 27. Takahashi Y, Imanaka T, Takano TSpatial and temporal pattern of smooth muscle cell differentiation during development of the vascular system in the mouse embryo. Anat Embryol (Berl) 1996;194:515–526.[PubMed][Google Scholar]
  • 28. Leveen P, et al Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes Dev. 1994;8:1875–1887.[PubMed][Google Scholar]
  • 29. Soriano PAbnormal kidney development and hematological disorders in PDGF beta-receptor mutant mice. Genes Dev. 1994;8:1888–1896.[PubMed][Google Scholar]
  • 30. Lindahl P, Johansson BR, Leveen P, Betsholtz CPericyte loss and microaneurysm formation in PDGF-B-deficient mice. Science. 1997;277:242–245.[PubMed][Google Scholar]
  • 31. Kuo CT, et al The LKLF transcription factor is required for normal tunica media formation and blood vessel stabilization during murine embryogenesis. Genes Dev. 1997;11:2996–3006.[Google Scholar]
  • 32. Wang F, et al. Sphingosine 1-phosphate stimulates cell migration through a G(i)-coupled cell surface receptor. Potential involvement in angiogenesis. J Biol Chem. 1999;274:35343–35350.[PubMed]
  • 33. Panetti TS, Nowlen J, Mosher DFSphingosine-1-phosphate and lysophosphatidic acid stimulate endothelial cell migration. Arterioscler Thromb Vasc Biol. 2000;20:1013–1019.[PubMed][Google Scholar]
  • 34. English D, et al Induction of endothelial cell chemotaxis by sphingosine 1-phosphate and stabilization of endothelial monolayer barrier function by lysophosphatidic acid, potential mediators of hematopoietic angiogenesis. J Hematother Stem Cell Res. 1999;8:627–634.[PubMed][Google Scholar]
  • 35. Kon J, et al Comparison of intrinsic activities of the putative sphingosine 1-phosphate receptor subtypes to regulate several signaling pathways in their cDNA-transfected Chinese hamster ovary cells. J Biol Chem. 1999;274:23940–23947.[PubMed][Google Scholar]
  • 36. Hall ARho GTPases and the actin cytoskeleton. Science. 1998;279:509–514.[PubMed][Google Scholar]
  • 37. Folkman J, D’Amore PABlood vessel formation: what is its molecular basis? Cell. 1996;87:1153–1155.[PubMed][Google Scholar]
  • 38. Hanahan DSignaling vascular morphogenesis and maintenance. Science. 1997;277:48–50.[PubMed][Google Scholar]
  • 39. Carmeliet P, et al Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature. 1996;380:435–439.[PubMed][Google Scholar]
  • 40. Ferrara N, et al Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature. 1996;380:439–442.[PubMed][Google Scholar]
  • 41. Shalaby F, et al Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature. 1995;376:62–66.[PubMed][Google Scholar]
  • 42. Fong GH, Rossant J, Gertsenstein M, Breitman MLRole of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature. 1995;376:66–70.[PubMed][Google Scholar]
  • 43. Dumont DJ, et al Dominant-negative and targeted null mutations in the endothelial receptor tyrosine kinase, tek, reveal a critical role in vasculogenesis of the embryo. Genes Dev. 1994;8:1897–1909.[PubMed][Google Scholar]
  • 44. Sato TN, et al Distinct roles of the receptor tyrosine kinases Tie-1 and Tie-2 in blood vessel formation. Nature. 1995;376:70–74.[PubMed][Google Scholar]
  • 45. Suri C, et al Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell. 1996;87:1171–1180.[PubMed][Google Scholar]
  • 46. Hellstrom M, Kal M, Lindahl P, Abramsson A, Betsholtz CRole of PDGF-B and PDGFR-beta in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. Development. 1999;126:3047–3055.[PubMed][Google Scholar]
  • 47. Shah NM, Groves AK, Anderson DJAlternative neural crest cell fates are instructively promoted by TGFbeta superfamily members. Cell. 1996;85:331–343.[PubMed][Google Scholar]
  • 48. Hirschi KK, Rohovsky SA, Beck LH, Smith SR, D’Amore PAEndothelial cells modulate the proliferation of mural cell precursors via platelet-derived growth factor-BB and heterotypic cell contact. Circ Res. 1999;84:298–305.[PubMed][Google Scholar]
  • 49. Hirschi KK, Rohovsky SA, D’Amore PA. PDGF, TGF-beta, and heterotypic cell-cell interactions mediate endothelial cell-induced recruitment of 10T1/2 cells and their differentiation to a smooth muscle fate [erratum, 1998. 141:1287] J Cell Biol. 1998;141:805–814.
  • 50. Li DY, et al Defective angiogenesis in mice lacking endoglin. Science. 1999;284:1534–1537.[PubMed][Google Scholar]
  • 51. Yang X, et al Angiogenesis defects and mesenchymal apoptosis in mice lacking SMAD5. Development. 1999;126:1571–1580.[PubMed][Google Scholar]
  • 52. Offermanns S, Mancino V, Revel JP, Simon MIVascular system defects and impaired cell chemokinesis as a result of Galpha13 deficiency. Science. 1997;275:533–536.[PubMed][Google Scholar]
  • 53. Lindahl P, Hellstrom M, Kalen M, Betsholtz CEndothelial-perivascular cell signaling in vascular development: lessons from knockout mice. Curr Opin Lipidol. 1998;9:407–411.[PubMed][Google Scholar]
  • 54. Nobes CD, Hall ARho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell. 1995;81:53–62.[PubMed][Google Scholar]
  • 55. Yatomi Y, Ruan F, Hakomori S, Igarashi YSphingosine-1-phosphate: a platelet-activating sphingolipid released from agonist-stimulated human platelets. Blood. 1995;86:193–202.[PubMed][Google Scholar]
  • 56. Yang L, Yatomi Y, Miura Y, Satoh K, Ozaki YMetabolism and functional effects of sphingolipids in blood cells. Br J Haematol. 1999;107:282–293.[PubMed][Google Scholar]
  • 57. Kupperman E, An S, Osborne N, Waldron S, Stainier DYRA sphingosine-1-phosphate receptor regulates cell migration during vertebrate heart development. Nature. 2000;406:192–195.[PubMed][Google Scholar]
  • 58. Lee O, et al Sphingosine 1-phosphate induces angiogenesis: its angiogenic action and signaling mechanism in human umbilical vein endothelial cells. Biochem Biophys Res Commun. 1999;264:743–750.[PubMed][Google Scholar]
  • 59. Folkman JAngiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med. 1995;1:27–31.[PubMed][Google Scholar]
  • 60. Pinedo HM, Verheul HM, D’Amato RJ, Folkman JInvolvement of platelets in tumour angiogenesis? Lancet. 1998;352:1775–1777.[PubMed][Google Scholar]
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.