Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Arteries and veins: making a difference with zebrafish

Key Points

  • One of the most fundamental distinctions between blood vessels is that between arteries and veins. These two types of vessel are structurally different and have long been functionally defined by the direction of the flow of blood that they carry. Recent work indicates that the endothelial cells that line the lumens of these two vessel types have distinct molecular identities.

  • Arterial–venous endothelial-cell identity is determined during embryogenesis, before circulation begins. Recent work in several vertebrates has begun to define the molecular pathways that specify this differentiated fate. The zebrafish has been particularly useful for in vivo dissection of the signalling pathways that regulate arterial and venous fate.

  • Zebrafish provide several advantages for studying vascular development. They are amenable to large-scale forward genetics analysis. The optical clarity, external embryonic development and small size of zebrafish embryos also allow easy visualization of the vasculature and allow screening for vascular-specific mutants.

  • Components of the Notch pathway are expressed in the blood vessels of many vertebrates, and several Notch receptors are expressed specifically in arterial endothelial cells. Studies in zebrafish have now shown that the activation of Notch signalling in endothelial cells promotes arterial cell fate and represses venous differentiation.

  • Studies in the zebrafish have also shown that the well-known signalling molecules — sonic hedgehog and vascular endothelial growth factor (Vegf) — act upstream of Notch signalling to promote arterial differentiation of endothelial cells. Several recent studies in the mouse have confirmed that Vegf also has an important role in arterial specification in this organism.

  • The powerful genetic tools provided by the zebrafish will allow both the identification of other molecules that are involved in arterial differentiation and the placement of these molecules in a genetic pathway. The conservation of signalling mechanisms and the similarity of embryonic and adult neovascularization indicate that the differentiation of arterial and venous endothelial cells will probably be relevant in the context of human disease.


Arteries and veins are structurally different and have long been functionally defined by the direction of blood flow that they carry. However, a growing body of evidence indicates that the identity of the endothelial cells that line these vessels is determined in the developing embryo, before circulation begins. Recent work on the zebrafish has led to the identification of signals that are responsible for arterial and venous differentiation of endothelial cells, and highlights the unique benefits of this model organism in the study of vascular development.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Molecular markers define arterial and venous cell identity.
Figure 2: The zebrafish vasculature.
Figure 3: Genetic analysis of signalling pathways.
Figure 4: A model for arterial differentiation in zebrafish.


  1. 1

    Folkman, J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nature Med. 1, 27–31 (1995).

    CAS  Article  Google Scholar 

  2. 2

    Cleaver, O. & Krieg, P. A. in Heart Development (eds Harvey, R. P. & Rosenthal, N.) 221–252 (Academic, San Diego, California, 1999).

    Book  Google Scholar 

  3. 3

    Risau, W. & Flamme, I. Vasculogenesis. Annu. Rev. Cell. Dev. Biol. 11, 73–91 (1995).

    CAS  Article  Google Scholar 

  4. 4

    Sabin, F. Studies on the origin of blood-vessels and of red blood corpuscles as seen in the living blastoderm of chicks on the second day of incubation. Contrib. Embryol. 36, 213–261 (1920).

    Google Scholar 

  5. 5

    Pardanaud, L., Yassine, F. & Dieterlen-Lievre, F. Relationship between vasculogenesis, angiogenesis and haemopoiesis during avian ontogeny. Development 105, 473–485 (1989).

    CAS  PubMed  Google Scholar 

  6. 6

    Poole, T. J. & Coffin, J. D. Vasculogenesis and angiogenesis: two distinct morphogenetic mechanisms establish embryonic vascular pattern. J. Exp. Zool. 251, 224–231 (1989).

    CAS  Article  Google Scholar 

  7. 7

    Wiltse, L. L. & Pait, T. G. Herophilus of Alexandria (325–255 B. C.). The father of anatomy. Spine 23, 1904–1914 (1998).

    CAS  Article  Google Scholar 

  8. 8

    Harvey, W. in Movement of the Heart and Blood in Animals; an Anatomical Essay, 209 (Blackwell Scientific, Oxford, UK, 1957).

    Google Scholar 

  9. 9

    Clark, E. R. Studies on the growth of blood vessels in the tail of the frog larvae. Am. J. Anat. 23, 37–88 (1918).

    Article  Google Scholar 

  10. 10

    Girard, H. Arterial pressure in the chick embryo. Am. J. Physiol. 224, 454–460 (1973).

    CAS  Article  Google Scholar 

  11. 11

    Gonzales-Crussi, F. Vasculogenesis in the chick embryo. An ultrastructural study. Am. J. Anat. 130, 441–460 (1971).

    Article  Google Scholar 

  12. 12

    Parry, E. W. & Abramovich, D. R. The ultrastructure of human umbilical vessel endothelium from early pregnancy to full term. J. Anat. 111, 29–42 (1972).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. 13

    Wang, H. U., Chen, Z. F. & Anderson, D. J. Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by efnB2 and its receptor Eph-B4. Cell 93, 741–753 (1998).This paper describes the molecular differences between arterial and venous endothelial cells that are apparent before the onset of circulation, and suggests the existence of genetic pathways that determine these cell types.

    CAS  Article  Google Scholar 

  14. 14

    Gerety, S. S., Wang, H. U., Chen, Z. F. & Anderson, D. J. Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand efnB2 in cardiovascular development. Mol. Cell 4, 403–414 (1999).

    CAS  Article  Google Scholar 

  15. 15

    Gale, N. W. et al. Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells. Dev. Biol. 230, 151–160 (2001).

    CAS  Article  Google Scholar 

  16. 16

    Shin, D. et al. Expression of ephrinB2 identifies a stable genetic difference between arterial and venous vascular smooth muscle as well as endothelial cells, and marks subsets of microvessels at sites of adult neovascularization. Dev. Biol. 230, 139–150 (2001).

    CAS  Article  Google Scholar 

  17. 17

    Zhong, T. P., Childs, S., Leu, J. P. & Fishman, M. C. Gridlock signalling pathway fashions the first embryonic artery. Nature 414, 216–220 (2001).This paper provides evidence that the specification of arterial and venous endothelial identity is an early step during vascular development.

    CAS  Article  Google Scholar 

  18. 18

    Moyon, D., Pardanaud, L., Yuan, L., Breant, C. & Eichmann, A. Plasticity of endothelial cells during arterial–venous differentiation in the avian embryo. Development 128, 3359–3370 (2001).

    CAS  PubMed  Google Scholar 

  19. 19

    Othman-Hassan, K. et al. Arterial identity of endothelial cells is controlled by local cues. Dev. Biol. 237, 398–409 (2001).

    CAS  Article  Google Scholar 

  20. 20

    Artavanis-Tsakonas, S., Rand, M. D. & Lake, R. J. Notch signaling: cell fate control and signal integration in development. Science 284, 770–776 (1999).

    CAS  Article  Google Scholar 

  21. 21

    Del Amo, F. F. et al. Expression pattern of Motch, a mouse homolog of Drosophila Notch, suggests an important role in early postimplantation mouse development. Development 115, 737–744 (1992).

    CAS  PubMed  Google Scholar 

  22. 22

    Zimrin, A. B. et al. An antisense oligonucleotide to the notch ligand jagged enhances fibroblast growth factor-induced angiogenesis in vitro. J. Biol. Chem. 271, 32499–32502 (1996).

    CAS  Article  Google Scholar 

  23. 23

    Uyttendaele, H. et al. Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene. Development 122, 2251–2259 (1996).

    CAS  PubMed  Google Scholar 

  24. 24

    Krebs, L. T. et al. Notch signaling is essential for vascular morphogenesis in mice. Genes Dev. 14, 1343–1352 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25

    Xue, Y. et al. Embryonic lethality and vascular defects in mice lacking the Notch ligand Jagged1. Hum. Mol. Genet. 8, 723–730 (1999).

    CAS  Article  Google Scholar 

  26. 26

    Uyttendaele, H., Ho, J., Rossant, J. & Kitajewski, J. Vascular patterning defects associated with expression of activated Notch4 in embryonic endothelium. Proc. Natl Acad. Sci. USA 98, 5643–5648 (2001).

    CAS  Article  Google Scholar 

  27. 27

    Lawson, N. D. et al. Notch signaling is required for arterial–venous differentiation during embryonic vascular development. Development 128, 3675–3683 (2001).This is among the first studies to provide definitive evidence of a genetic component that determines the arterial or venous identity of blood vessels.

    CAS  Google Scholar 

  28. 28

    Villa, N. et al. Vascular expression of Notch pathway receptors and ligands is restricted to arterial vessels. Mech. Dev. 108, 161–164 (2001).

    CAS  Article  Google Scholar 

  29. 29

    Smithers, L., Haddon, C., Jiang, Y. & Lewis, J. Sequence and embryonic expression of deltaC in the zebrafish. Mech. Dev. 90, 119–123 (2000).

    CAS  Article  Google Scholar 

  30. 30

    Jiang, Y. J. et al. Mutations affecting neurogenesis and brain morphology in the zebrafish, Danio rerio. Development 123, 205–216 (1996).

    CAS  Google Scholar 

  31. 31

    Wettstein, D. A., Turner, D. L. & Kintner, C. The Xenopus homolog of Drosophila Suppressor of Hairless mediates Notch signaling during primary neurogenesis. Development 124, 693–702 (1997).

    CAS  PubMed  Google Scholar 

  32. 32

    Lawson, N. D., Vogel, A. M. & Weinstein, B. M. Sonic hedgehog and vascular endothelial growth factor act upstream of the Notch pathway during arterial endothelial differentiation. Dev. Cell 3, 127–136 (2002).This study shows the suitability of the zebrafish for analysing signalling pathways in a series of epistasis experiments that define a genetic pathway that is responsible for arterial differentiation.

    CAS  Article  Google Scholar 

  33. 33

    Thompson, M. A. et al. The cloche and spadetail genes differentially affect hematopoiesis and vasculogenesis. Dev. Biol. 197, 248–269 (1998).

    CAS  Article  Google Scholar 

  34. 34

    Adams, R. H. et al. Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev. 13, 295–306 (1999).

    CAS  Article  Google Scholar 

  35. 35

    Helbling, P. M., Saulnier, D. M. & Brandli, A. W. The receptor tyrosine kinase EphB4 and ephrin-B ligands restrict angiogenic growth of embryonic veins in Xenopus laevis. Development 127, 269–278 (2000).

    CAS  PubMed  Google Scholar 

  36. 36

    Stainier, D. Y. et al. Mutations affecting the formation and function of the cardiovascular system in the zebrafish embryo. Development 123, 285–292 (1996).

    CAS  Google Scholar 

  37. 37

    Weinstein, B. M., Stemple, D. L., Driever, W. & Fishman, M. C. Gridlock, a localized heritable vascular patterning defect in the zebrafish. Nature Med. 1, 1143–1147 (1995).

    CAS  Article  Google Scholar 

  38. 38

    Zhong, T. P., Rosenberg, M., Mohideen, M. A., Weinstein, B. & Fishman, M. C. gridlock, an HLH gene required for assembly of the aorta in zebrafish. Science 287, 1820–1824 (2000).

    CAS  Article  Google Scholar 

  39. 39

    Nakagawa, O. et al. Members of the HRT family of basic helix–loop–helix proteins act as transcriptional repressors downstream of notch signaling. Proc. Natl Acad. Sci. USA 97, 13655–13660 (2000).

    CAS  Article  Google Scholar 

  40. 40

    Roelink, H. et al. Floor plate and motor neuron induction by different concentrations of the amino-terminal cleavage product of sonic hedgehog autoproteolysis. Cell 81, 445–455 (1995).

    CAS  Article  Google Scholar 

  41. 41

    Ericson, J., Morton, S., Kawakami, A., Roelink, H. & Jessell, T. M. Two critical periods of Sonic Hedgehog signaling required for the specification of motor neuron identity. Cell 87, 661–673 (1996).

    CAS  Article  Google Scholar 

  42. 42

    Fan, C. M. & Tessier-Lavigne, M. Patterning of mammalian somites by surface ectoderm and notochord: evidence for sclerotome induction by a hedgehog homolog. Cell 79, 1175–1186 (1994).

    CAS  Article  Google Scholar 

  43. 43

    Schauerte, H. E. et al. Sonic hedgehog is not required for the induction of medial floor plate cells in the zebrafish. Development 125, 2983–2993 (1998).

    CAS  PubMed  Google Scholar 

  44. 44

    van Eeden, F. J. et al. Mutations affecting somite formation and patterning in the zebrafish, Danio rerio. Development 123, 153–164 (1996).

    CAS  Google Scholar 

  45. 45

    Chen, J. N. et al. Mutations affecting the cardiovascular system and other internal organs in zebrafish. Development 123, 293–302 (1996).

    CAS  PubMed  Google Scholar 

  46. 46

    Brown, L. A. et al. Insights into early vasculogenesis revealed by expression of the ETS-domain transcription factor Fli-1 in wild-type and mutant zebrafish embryos. Mech. Dev. 90, 237–252 (2000).

    CAS  Article  Google Scholar 

  47. 47

    Carmeliet, P. et al. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380, 435–439 (1996).

    CAS  Article  Google Scholar 

  48. 48

    Stalmans, I. et al. Arteriolar and venular patterning in retinas of mice selectively expressing VEGF isoforms. J. Clin. Invest. 109, 327–336 (2002).One of the first studies to show that Vegf is specifically required for arterial, but not venous, blood vessel development.

    CAS  Article  Google Scholar 

  49. 49

    Mukouyama, Y., Shin, D., Britsch, S., Taniguchi, M. & Anderson, D. J. Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin. Cell 109, 693–705 (2002).The authors provide convincing evidence that Vegf is necessary for arterial differentiation and show that Vegf can directly induce artery marker gene expression without affecting endothelial cell proliferation or survival.

    CAS  Article  Google Scholar 

  50. 50

    Visconti, R. P., Richardson, C. D. & Sato, T. N. Orchestration of angiogenesis and arteriovenous contribution by angiopoietins and vascular endothelial growth factor (VEGF). Proc. Natl Acad. Sci. USA 99, 8219–8224 (2002).

    CAS  Article  Google Scholar 

  51. 51

    Pola, R. et al. The morphogen Sonic hedgehog is an indirect angiogenic agent upregulating two families of angiogenic growth factors. Nature Med. 7, 706–711 (2001).

    CAS  Article  Google Scholar 

  52. 52

    Soker, S., Takashima, S., Miao, H. Q., Neufeld, G. & Klagsbrun, M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92, 735–745 (1998).

    CAS  Article  Google Scholar 

  53. 53

    Ahn, D. G., Ruvinsky, I., Oates, A. C., Silver, L. M. & Ho, R. K. tbx20, a new vertebrate T-box gene expressed in the cranial motor neurons and developing cardiovascular structures in zebrafish. Mech. Dev. 95, 253–258 (2000).

    CAS  Article  Google Scholar 

  54. 54

    Shutter, J. R. et al. Dll4, a novel Notch ligand expressed in arterial endothelium. Genes Dev. 14, 1313–1318 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. 55

    Kudoh, T. et al. A gene expression screen in zebrafish embryogenesis. Genome Res. 11, 1979–1987 (2001).

    CAS  Article  Google Scholar 

  56. 56

    Urness, L. D., Sorensen, L. K. & Li, D. Y. Arteriovenous malformations in mice lacking activin receptor-like kinase-1. Nature Genet. 26, 328–331 (2000).

    CAS  Article  Google Scholar 

  57. 57

    Roman, B. L. et al. Disruption of acvrl1 increases endothelial cell number in zebrafish cranial vessels. Development 129, 3009–3019 (2002).

    CAS  PubMed  Google Scholar 

  58. 58

    Odenthal, J. et al. Mutations affecting the formation of the notochord in the zebrafish, Danio rerio. Development 123, 103–115 (1996).

    CAS  PubMed  Google Scholar 

  59. 59

    Fouquet, B., Weinstein, B. M., Serluca, F. C. & Fishman, M. C. Vessel patterning in the embryo of the zebrafish: guidance by notochord. Dev. Biol. 183, 37–48 (1997).

    CAS  Article  Google Scholar 

  60. 60

    Orioli, D. & Klein, R. The Eph receptor family: axonal guidance by contact repulsion. Trends Genet. 13, 354–359 (1997).

    CAS  Article  Google Scholar 

  61. 61

    Isogai, S., Horiguchi, M. & Weinstein, B. M. The vascular anatomy of the developing zebrafish: an atlas of embryonic and early larval development. Dev. Biol. 230, 278–301 (2001).

    CAS  Article  Google Scholar 

  62. 62

    Motoike, T. et al. Universal GFP reporter for the study of vascular development. Genesis 28, 75–81 (2000).

    CAS  Article  Google Scholar 

  63. 63

    Lawson, N. D. & Weinstein, B. M. In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev. Biol. 249, 307–318 (2002).

    Article  Google Scholar 

  64. 64

    Liang, D. et al. Cloning and characterization of vascular endothelial growth factor (VEGF) from zebrafish, Danio rerio. Biochim. Biophys. Acta 1397, 14–20 (1998).

    CAS  Article  Google Scholar 

  65. 65

    Scheer, N., Groth, A., Hans, S. & Campos-Ortega, J. A. An instructive function for Notch in promoting gliogenesis in the zebrafish retina. Development 128, 1099–1107 (2001).

    CAS  Google Scholar 

  66. 66

    Scheer, N. & Campos-Ortega, J. A. Use of the Gal4–UAS technique for targeted gene expression in the zebrafish. Mech. Dev. 80, 153–158 (1999).

    CAS  Article  Google Scholar 

  67. 67

    Nasevicius, A., Larson, J. & Ekker, S. C. Distinct requirements for zebrafish angiogenesis revealed by a VEGF-A morphant. Yeast 17, 294–301 (2000).

    CAS  Article  Google Scholar 

  68. 68

    Liao, W. et al. The zebrafish gene cloche acts upstream of a flk-1 homologue to regulate endothelial cell differentiation. Development 124, 381–389 (1997).

    CAS  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Brant M. Weinstein.

Related links

Related links

















haemorrhagic telangiectasia type II













Dawid lab in situ screen

Interactive Atlas of Zebrafish Vascular Anatomy home page

Thisse lab in situ screen

Weinstein lab



Consists of mesodermal cells that, after gastrulation, lie between the ectodermal and endodermal layers in vertebrate embryos. These cells give rise to haematopoietic, vascular and kidney tissue.


De novo formation of blood vessels through the coalescence of endothelial cells. It often involves extensive migration of endothelial cells from their point of origin to the site of vessel formation.


The formation of new blood vessels from pre-existing ones. It is often associated with cell division and the subsequent sprouting of the endothelial cells that contribute to the growing blood vessel.


In mouse, the extra-embryonic tissue that surrounds the yolk.


An undifferentiated endothelial progenitor cell that has yet to integrate into a blood vessel.


The notochord and its anterior extension, the prechordal plate.


An abnormal connection between a main artery and vein, such as the dorsal aorta and posterior cardinal vein, that leads to a circulatory bypass.


A chemically modified antisense oligonucleotide that can specifically inhibit translation of a target mRNA.


A transient structure that is located at the midline in the trunk of developing vertebrate embryos.


Presumptive spinal cord.


The segmental blocks of mesenchyme that are adjacent to the notochord and that give rise to the muscle tissue of the trunk.


A small-calibre artery that is continuous with a capillary network and that is associated with only one or two layers of surrounding smooth muscle cells.


A small-calibre vein that is continuous with a capillary network.


A light microscope that allows imaging of fluorescent structures in thick (tens to hundreds of micrometres) specimens. A series of optical 'slices' are collected using a scanning laser beam and specially designed optics to eliminate out-of-focus excited fluorescence. The slices are reconstructed to provide detailed 3D representations of the image data.


A dilation of the small vessels in capillary beds that often leads to hyperpermeability and haemorrhage.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Lawson, N., Weinstein, B. Arteries and veins: making a difference with zebrafish. Nat Rev Genet 3, 674–682 (2002).

Download citation

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing