Anatomical Science International

, Volume 84, Issue 3, pp 95–101 | Cite as

Regulation of endothelial cell differentiation and arterial specification by VEGF and Notch signaling

Special Issue on Cardiovascular Development Review Article


Analysis of molecular and cellular mechanisms underlying vascular development in vertebrates indicates that initially vasculogenesis occurs when a primary capillary plexus forms de novo from endothelial cell precursors derived from nascent mesodermal cells. Transplantation experiments in avian embryos demonstrate that embryonic endothelial cells originate from two different mesodermal lineages: splanchnic mesoderm and somites. Genetic analysis of mouse and zebrafish reveals that vascular endothelial growth factor (VEGF)/Flk1 and Notch signaling play crucial roles throughout embryonic vascular development. VEGFA plays a major role in endothelial cell proliferation, migration, survival, and regulation of vascular permeability. Flk1, the primary VEGFA receptor, is the earliest marker of the developing endothelial lineage and is essential for endothelial differentiation during vasculogenesis. Notch signaling has been demonstrated to directly induce arterial endothelial differentiation. Recent studies suggest that Notch signaling is activated downstream of VEGF signaling and negatively regulates VEGF-induced angiogenesis and suppresses aberrant vascular branching morphogenesis. In addition to altering endothelial cell fate through Notch activation, VEGFA directly guides endothelial cell migration in an isoform-dependent manner, modifying vascular patterns. Interestingly, genetic studies in mice show that many molecules involved in VEGF or Notch signaling must be tightly regulated for proper vascular formation. Taken together, VEGF and Notch signaling apparently coordinate vascular patterning by regulating each other.


Embryo Endothelial cells Notch Vascular development VEGF 


  1. Adams RH, Alitalo K (2007) Molecular regulation of angiogenesis and lymphangiogenesis. Nat Rev Mol Cell Biol 8:464–478PubMedCrossRefGoogle Scholar
  2. Ambler CA, Nowicki JL, Burke AC, Bautch VL (2001) Assembly of trunk and limb blood vessels involves extensive migration and vasculogenesis of somite-derived angioblasts. Dev Biol 234:352–364PubMedCrossRefGoogle Scholar
  3. Carmeliet P, Ferreira V, Breier G et al (1996) Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380:435–439PubMedCrossRefGoogle Scholar
  4. Carmeliet P, Moons L, Luttun A et al (2001) Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions. Nat Med 7:575–583PubMedCrossRefGoogle Scholar
  5. Choi K, Kennedy M, Kazarov A, Papadimitriou JC, Keller G (1998) A common precursor for hematopoietic and endothelial cells. Development 125:725–732PubMedGoogle Scholar
  6. Covassin LD, Villefranc JA, Kacergis MC, Weinstein BM, Lawson ND (2006) Distinct genetic interactions between multiple Vegf receptors are required for development of different blood vessel types in zebrafish. Proc Natl Acad Sci USA 103:6554–6559PubMedCrossRefGoogle Scholar
  7. Duarte A, Hirashima M, Benedito R et al (2004) Dosage-sensitive requirement for mouse Dll4 in artery development. Genes Dev 18:2474–2478PubMedCrossRefGoogle Scholar
  8. Dumont DJ, Jussila L, Taipale J et al (1998) Cardiovascular failure in mouse embryos deficient in VEGF receptor-3. Science 282:946–949PubMedCrossRefGoogle Scholar
  9. Ferrara N, Carver Moore K, Chen H et al (1996) Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature 380:439–442PubMedCrossRefGoogle Scholar
  10. Fong GH, Rossant J, Gertsenstein M, Breitman ML (1995) Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376:66–70PubMedCrossRefGoogle Scholar
  11. Gale NW, Dominguez MG, Noguera I et al (2004) Haploinsufficiency of delta-like 4 ligand results in embryonic lethality due to major defects in arterial and vascular development. Proc Natl Acad Sci USA 101:15949–15954PubMedCrossRefGoogle Scholar
  12. Gerhardt H, Golding M, Fruttiger M et al (2003) VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. J Cell Biol 161:1163–1177PubMedCrossRefGoogle Scholar
  13. Hellstrom M, Phng LK, Hofmann JJ et al (2007) Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis. Nature 445:776–780PubMedCrossRefGoogle Scholar
  14. Hidaka M, Stanford WL, Bernstein A (1999) Conditional requirement for the Flk-1 receptor in the in vitro generation of early hematopoietic cells. Proc Natl Acad Sci USA 96:7370–7375PubMedCrossRefGoogle Scholar
  15. Hiratsuka S, Minowa O, Kuno J, Noda T, Shibuya M (1998) Flt-1 lacking the tyrosine kinase domain is sufficient for normal development and angiogenesis in mice. Proc Natl Acad Sci USA 95:9349–9354PubMedCrossRefGoogle Scholar
  16. Krebs LT, Xue Y, Norton CR et al (2000) Notch signaling is essential for vascular morphogenesis in mice. Genes Dev 14:1343–1352PubMedGoogle Scholar
  17. Krebs LT, Shutter JR, Tanigaki K, Honjo T, Stark KL, Gridley T (2004) Haploinsufficient lethality and formation of arteriovenous malformations in Notch pathway mutants. Genes Dev 18:2469–2473PubMedCrossRefGoogle Scholar
  18. Lawson ND, Scheer N, Pham VN et al (2001) Notch signaling is required for arterial-venous differentiation during embryonic vascular development. Development 128:3675–3683PubMedGoogle Scholar
  19. Lawson ND, Vogel AM, Weinstein BM (2002) Sonic hedgehog and vascular endothelial growth factor act upstream of the Notch pathway during arterial endothelial differentiation. Dev Cell 3:127–136PubMedCrossRefGoogle Scholar
  20. Lawson ND, Mugford JW, Diamond BA, Weinstein BM (2003) Phospholipase C gamma-1 is required downstream of vascular endothelial growth factor during arterial development. Genes Dev 17:1346–1351PubMedCrossRefGoogle Scholar
  21. Le Douarin NM (1974) Cell recognition based on natural morphological nuclear markers. Med Biol 52:281–319PubMedGoogle Scholar
  22. Lee S, Jilani SM, Nikolova GV, Carpizo D, Iruela-Arispe ML (2005) Processing of VEGF-A by matrix metalloproteinases regulates bioavailability and vascular patterning in tumors. J Cell Biol 169:681–691PubMedCrossRefGoogle Scholar
  23. Lee S, Chen TT, Barber CL et al (2007) Autocrine VEGF signaling is required for vascular homeostasis. Cell 130:691–703PubMedCrossRefGoogle Scholar
  24. Limbourg A, Ploom M, Elligsen D et al (2007) Notch ligand Delta-like 1 is essential for postnatal arteriogenesis. Circ Res 100:363–371PubMedCrossRefGoogle Scholar
  25. Liu ZJ, Shirakawa T, Li Y et al (2003) Regulation of Notch1 and Dll4 by vascular endothelial growth factor in arterial endothelial cells: implications for modulating arteriogenesis and angiogenesis. Mol Cell Biol 23:14–25PubMedCrossRefGoogle Scholar
  26. Matsumura K, Hirashima M, Ogawa M et al (2003) Modulation of VEGFR-2-mediated endothelial-cell activity by VEGF-C/VEGFR-3. Blood 101:1367–1374PubMedCrossRefGoogle Scholar
  27. Millauer B, Wizigmann-Voos S, Schnurch H et al (1993) High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell 72:835–846PubMedCrossRefGoogle Scholar
  28. Miquerol L, Langille BL, Nagy A (2000) Embryonic development is disrupted by modest increases in vascular endothelial growth factor gene expression. Development 127:3941–3946PubMedGoogle Scholar
  29. Mukouyama YS, Shin D, Britsch S, Taniguchi M, Anderson DJ (2002) Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin. Cell 109:693–705PubMedCrossRefGoogle Scholar
  30. Mukouyama YS, Gerber HP, Ferrara N, Gu C, Anderson DJ (2005) Peripheral nerve-derived VEGF promotes arterial differentiation via neuropilin 1-mediated positive feedback. Development 132:941–952PubMedCrossRefGoogle Scholar
  31. Ng YS, Rohan R, Sunday ME, Demello DE, D’Amore PA (2001) Differential expression of VEGF isoforms in mouse during development and in the adult. Dev Dyn 220:112–121PubMedCrossRefGoogle Scholar
  32. Nimmagadda S, Geetha-Loganathan P, Scaal M, Christ B, Huang R (2007) FGFs, Wnts and BMPs mediate induction of VEGFR-2 (Quek-1) expression during avian somite development. Dev Biol 305:421–429PubMedCrossRefGoogle Scholar
  33. Nishikawa SI, Nishikawa S, Hirashima M, Matsuyoshi N, Kodama H (1998) Progressive lineage analysis by cell sorting and culture identifies FLK1+VE-cadherin+ cells at a diverging point of endothelial and hemopoietic lineages. Development 125:1747–1757PubMedGoogle Scholar
  34. Noguera-Troise I, Daly C, Papadopoulos NJ et al (2006) Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis. Nature 444:1032–1037PubMedCrossRefGoogle Scholar
  35. Oliver G, Alitalo K (2005) The lymphatic vasculature: recent progress and paradigms. Annu Rev Cell Dev Biol 21:457–483PubMedCrossRefGoogle Scholar
  36. Pardanaud L, Altmann C, Kitos P, Dieterlen-Lievre F, Buck CA (1987) Vasculogenesis in the early quail blastodisc as studied with a monoclonal antibody recognizing endothelial cells. Development 100:339–349PubMedGoogle Scholar
  37. Pardanaud L, Luton D, Prigent M, Bourcheix LM, Catala M, Dieterlen-Lievre F (1996) Two distinct endothelial lineages in ontogeny, one of them related to hemopoiesis. Development 122:1363–1371PubMedGoogle Scholar
  38. Park JE, Keller GA, Ferrara N (1993) The vascular endothelial growth factor (VEGF) isoforms: differential deposition into the subepithelial extracellular matrix and bioactivity of extracellular matrix-bound VEGF. Mol Biol Cell 4:1317–1326PubMedGoogle Scholar
  39. Peault BM, Thiery JP, Le Douarin NM (1983) Surface marker for hemopoietic and endothelial cell lineages in quail that is defined by a monoclonal antibody. Proc Natl Acad Sci USA 80:2976–2980PubMedCrossRefGoogle Scholar
  40. Pouget C, Gautier R, Teillet MA, Jaffredo T (2006) Somite-derived cells replace ventral aortic hemangioblasts and provide aortic smooth muscle cells of the trunk. Development 133:1013–1022PubMedCrossRefGoogle Scholar
  41. Ridgway J, Zhang G, Wu Y et al (2006) Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature 444:1083–1087PubMedCrossRefGoogle Scholar
  42. Rossant J, Hirashima M (2003) Vascular development and patterning: making the right choices. Curr Opin Genet Dev 13:408–412PubMedCrossRefGoogle Scholar
  43. Ruhrberg C (2003) Growing and shaping the vascular tree: multiple roles for VEGF. Bioessays 25:1052–1060PubMedCrossRefGoogle Scholar
  44. Ruhrberg C, Gerhardt H, Golding M et al (2002) Spatially restricted patterning cues provided by heparin-binding VEGF-A control blood vessel branching morphogenesis. Genes Dev 16:2684–2698PubMedCrossRefGoogle Scholar
  45. Sabin FR (1920) Studies on the origin of blood vessels and of red corpuscles as seen in the living blastoderm of the chick during the second day of incubation. Contrib Embryol 9:213–262Google Scholar
  46. Sakurai Y, Ohgimoto K, Kataoka Y, Yoshida N, Shibuya M (2005) Essential role of Flk-1 (VEGF receptor 2) tyrosine residue 1173 in vasculogenesis in mice. Proc Natl Acad Sci USA 102:1076–1081PubMedCrossRefGoogle Scholar
  47. Schuh AC, Faloon P, Hu QL, Bhimani M, Choi K (1999) In vitro hematopoietic and endothelial potential of flk-1(−/−) embryonic stem cells and embryos. Proc Natl Acad Sci USA 96:2159–2164PubMedCrossRefGoogle Scholar
  48. Seo S, Fujita H, Nakano A, Kang M, Duarte A, Kume T (2006) The forkhead transcription factors, Foxc1 and Foxc2, are required for arterial specification and lymphatic sprouting during vascular development. Dev Biol 294:458–470PubMedCrossRefGoogle Scholar
  49. Shalaby F, Rossant J, Yamaguchi TP et al (1995) Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 376:62–66PubMedCrossRefGoogle Scholar
  50. Shalaby F, Ho J, Stanford WL et al (1997) A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. Cell 89:981–990PubMedCrossRefGoogle Scholar
  51. Siekmann AF, Lawson ND (2007) Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries. Nature 445:781–784PubMedCrossRefGoogle Scholar
  52. Takashima S, Kitakaze M, Asakura M et al (2002) Targeting of both mouse neuropilin-1 and neuropilin-2 genes severely impairs developmental yolk sac and embryonic angiogenesis. Proc Natl Acad Sci USA 99:3657–3662PubMedCrossRefGoogle Scholar
  53. Tammela T, Zarkada G, Wallgard E et al (2008) Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation. Nature 454:656–660PubMedCrossRefGoogle Scholar
  54. Torres-Vazquez J, Kamei M, Weinstein BM (2003) Molecular distinction between arteries and veins. Cell Tissue Res 314:43–59PubMedCrossRefGoogle Scholar
  55. Ueno H, Weissman IL (2006) Clonal analysis of mouse development reveals a polyclonal origin for yolk sac blood islands. Dev Cell 11:519–533PubMedCrossRefGoogle Scholar
  56. Uyttendaele H, Ho J, Rossant J, Kitajewski J (2001) Vascular patterning defects associated with expression of activated Notch4 in embryonic endothelium. Proc Natl Acad Sci USA 98:5643–5648PubMedCrossRefGoogle Scholar
  57. Visconti RP, Richardson CD, Sato TN (2002) Orchestration of angiogenesis and arteriovenous contribution by angiopoietins and vascular endothelial growth factor (VEGF). Proc Natl Acad Sci USA 99:8219–8224PubMedCrossRefGoogle Scholar
  58. Vogeli KM, Jin SW, Martin GR, Stainier DY (2006) A common progenitor for haematopoietic and endothelial lineages in the zebrafish gastrula. Nature 443:337–339PubMedCrossRefGoogle Scholar
  59. Williams CK, Li JL, Murga M, Harris AL, Tosato G (2006) Up-regulation of the Notch ligand Delta-like 4 inhibits VEGF-induced endothelial cell function. Blood 107:931–939PubMedCrossRefGoogle Scholar
  60. Yamaguchi TP, Dumont DJ, Conlon RA, Breitman ML, Rossant J (1993) flk-1, an flt-related receptor tyrosine kinase is an early marker for endothelial cell precursors. Development 118:489–498PubMedGoogle Scholar

Copyright information

© Japanese Association of Anatomists 2009

Authors and Affiliations

  1. 1.Division of Vascular Biology, Department of Physiology and Cell BiologyKobe University Graduate School of MedicineKobeJapan

Personalised recommendations