Molecular and Cellular Mechanisms of Vascular Development


Blood vessels form an extensive organ system that enables gas exchange, delivers nutrients and removes waste products from tissues and is, therefore, essential for vertebrate life. Blood vessels additionally regulate leukocyte trafficking to support immune system function and transport endocrine hormones for the systemic regulation of physiological processes. During embryonic development, blood vessels also secrete cues that regulate the formation of other organs. To ensure that functional vasculature forms in the embryo, a large number of molecules regulate a wide range of cellular mechanisms that collectively act on the endothelial cells that form the inner lining of all blood vessels. These molecular and cellular mechanisms may be reactivated after birth to induce blood vessel growth that is beneficial by countering tissue ischemia or pathological if excessive or dysfunctional. Understanding developmental blood vessel growth, therefore, provides knowledge that may be used to develop new therapeutic strategies for a range of diseases.

Key Concepts

  • The circulatory system is the first organ to form during vertebrate embryogenesis and is required for the subsequent development of other organs.
  • The circulatory system is the first organ to form during vertebrate embryogenesis.
  • Proper blood vessel growth and remodeling is required for the subsequent development of other organs.
  • Blood vessel development can be studied using the embryonic mouse hindbrain and perinatal mouse retina.
  • Many molecular pathways synergise to regulate blood vessel growth.
  • Elucidating the mechanisms of vascular development may advance novel therapies to treat vascular insufficiency in ischemic diseases.

Keywords: endothelial cell; angiogenesis; vasculogenesis; angioblast; erythromyeloid progenitor; neural progenitor cell; retina; brain; lung; liver; oxygen‐induced retinopathy

Figure 1. Mechanisms of blood vessel growth. (a) Vasculogenesis involves the differentiation of angioblasts into ECs and their coalescence into lumenised vascular structure. (b) Sprouting angiogenesis involves the formation of EC tip cells to drive the extension of new vessel segments from pre‐existing vessels. (c) Intussusceptive angiogenesis occurs when a blood vessel splits into two after ECs migrate to the centre of the vascular lumen to form a pillar that causes the vessel to split. (d) EMP‐derived progenitors in the circulation can insert into the vascular wall to become ECs and thereby extend the vascular surface.
Figure 2. Mouse models of developmental and pathological angiogenesis (a,b) Hindbrain angiogenesis model: (a) Schematic representation of a hindbrain dissection from an E12.5 mouse embryo. (b) Fluorescent micrograph of the SVP of an E12.5 hindbrain labelled with the EC marker isolectin B4 (IB4); scale bar 250 μm. The boxed area is shown at higher magnification on the right‐hand side; scale bar 50 μm. (c,d) Retina angiogenesis model: (c) Schematic representation of a retina dissection from a P7 mouse eye. (d) Fluorescent micrograph of a P7 retina labelled with IB4; scale bar 1 mm. The boxed area is shown at higher magnification on the right‐hand side; scale bar 100 μm. (e,f) OIR neoangiogenesis model: (e) Schematic representation of hyperoxia‐induced vaso‐obliteration on P12 and the subsequent formation of vascular tufts after return to normoxia pn P17. (f) Fluorescent micrograph of a P17 retina labelled with IB4; scale bar 1 mm. The boxed area is shown at higher magnification on the right‐hand side; scale bar 100 μm.


Aspalter IM, Gordon E, Dubrac A, et al. (2015) Alk1 and Alk5 inhibition by Nrp1 controls vascular sprouting downstream of Notch. Nature Communications 6: 7264.

Bellusci S, Furuta Y, Rush MG, et al. (1997) Involvement of Sonic hedgehog (Shh) in mouse embryonic lung growth and morphogenesis. Development 124: 53–63.

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

Carmeliet P and Jain RK (2011) Molecular mechanisms and clinical applications of angiogenesis. Nature 473: 298–307.

Connolly SE, Hores TA, Smith LE and D'amore PA (1988) Characterization of vascular development in the mouse retina. Microvascular Research 36: 275–290.

Crafts TD, Jensen AR, Blocher‐Smith EC and Markel TA (2015) Vascular endothelial growth factor: therapeutic possibilities and challenges for the treatment of ischemia. Cytokine 71: 385–393.

De Bock K, Georgiadou M, Schoors S, et al. (2013) Role of PFKFB3‐driven glycolysis in vessel sprouting. Cell 154: 651–663.

Del Moral PM, Sala FG, Tefft D, et al. (2006) VEGF‐A signaling through Flk‐1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis. Developmental Biology 290: 177–188.

De Spiegelaere W, Casteleyn C, Van Den Broeck W, et al. (2012) Intussusceptive angiogenesis: a biologically relevant form of angiogenesis. Journal of Vascular Research 49: 390–404.

Duarte A, Hirashima M, Benedito R, et al. (2004) Dosage‐sensitive requirement for mouse Dll4 in artery development. Genes & Development 18: 2474–2478.

Eelen G, De Zeeuw P, Treps L, et al. (2018) Endothelial cell metabolism. Physiological Reviews 98: 3–58.

Fantin A, Vieira JM, Gestri G, et al. (2010) Tissue macrophages act as cellular chaperones for vascular anastomosis downstream of VEGF‐mediated endothelial tip cell induction. Blood 116: 829–840.

Fantin A, Vieira JM, Plein A, Maden CH and Ruhrberg C (2013) The embryonic mouse hindbrain as a qualitative and quantitative model for studying the molecular and cellular mechanisms of angiogenesis. Nature Protocols 8: 418–429.

Fantin A, Herzog B, Mahmoud M, et al. (2014) Neuropilin 1 (NRP1) hypomorphism combined with defective VEGF‐A binding reveals novel roles for NRP1 in developmental and pathological angiogenesis. Development 141: 556–562.

Fantin A, Lampropoulou A, Gestri G, et al. (2015) NRP1 regulates CDC42 activation to promote filopodia formation in endothelial tip cells. Cell Reports 11: 1577–1590.

Ferrara N, Carver‐Moore K, Chen H, et al. (1996) Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature 380: 439–442.

Fish JE and Wythe JD (2015) The molecular regulation of arteriovenous specification and maintenance. Developmental Dynamics 244: 391–409.

Fong G‐H, Rossant J, Gertsenstein M and Breitman ML (1995) Role of the Flt‐1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376: 66–70.

Fong GH, Zhang L, Bryce DM and Peng J (1999) Increased hemangioblast commitment, not vascular disorganization, is the primary defect in flt‐1 knock‐out mice. Development (Cambridge, England) 126: 3015–3025.

Fruttiger M (2007) Development of the retinal vasculature. Angiogenesis 10: 77–88.

Gelfand MV, Hagan N, Tata A, et al. (2014) Neuropilin‐1 functions as a VEGFR2 co‐receptor to guide developmental angiogenesis independent of ligand binding. eLife 3: e03720.

Gerber HP, Mcmurtrey A, Kowalski J, et al. (1998) Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3'‐kinase/Akt signal transduction pathway. Requirement for Flk‐1/KDR activation. The Journal of Biological Chemistry 273: 30336–30343.

Gerhardt H, Golding M, Fruttiger M, et al. (2003) VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. The Journal of Cell Biology 161: 1163–1177.

Geudens I and Gerhardt H (2011) Coordinating cell behaviour during blood vessel formation. Development 138: 4569–4583.

Ginhoux F and Guilliams M (2016) Tissue‐resident macrophage ontogeny and homeostasis. Immunity 44: 439–449.

Herriges M and Morrisey EE (2014) Lung development: orchestrating the generation and regeneration of a complex organ. Development 141: 502–513.

Hillman RT, Feng BY, Ni J, et al. (2011) Neuropilins are positive regulators of Hedgehog signal transduction. Genes & Development 25: 2333–2346.

Hirota S, Clements TP, Tang LK, et al. (2015) Neuropilin 1 balances beta8 integrin‐activated TGFbeta signaling to control sprouting angiogenesis in the brain. Development 142: 4363–4373.

Iruela‐Arispe ML and Davis GE (2009) Cellular and molecular mechanisms of vascular lumen formation. Developmental Cell 16: 222–231.

Ito M and Yoshioka M (1999) Regression of the hyaloid vessels and pupillary membrane of the mouse. Anatomy and Embryology 200: 403–411.

Jain RK (2005) Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307: 58–62.

Kawasaki T, Kitsukawa T, Bekku Y, et al. (1999) A requirement for neuropilin‐1 in embryonic vessel formation. Development 126: 4895–4902.

Kim LA and D'amore PA (2012) A brief history of anti‐VEGF for the treatment of ocular angiogenesis. The American Journal of Pathology 181: 376–379.

Lanahan A, Zhang X, Fantin A, et al. (2013) The neuropilin 1 cytoplasmic domain is required for VEGF‐A‐dependent arteriogenesis. Developmental Cell 25: 156–168.

Lange C, Turrero Garcia M, Decimo I, et al. (2016) Relief of hypoxia by angiogenesis promotes neural stem cell differentiation by targeting glycolysis. The EMBO Journal 35: 924–941.

Lawson ND, Scheer N, Pham VN, et al. (2001) Notch signaling is required for arterial‐venous differentiation during embryonic vascular development. Development (Cambridge, England) 128: 3675–3683.

Lecouter J, Moritz DR, Li B, et al. (2003) Angiogenesis‐independent endothelial protection of liver: role of VEGFR‐1. Science 299: 890–893.

Matsumoto K, Yoshitomi H, Rossant J and Zaret KS (2001) Liver organogenesis promoted by endothelial cells prior to vascular function. Science 294: 559–563.

Mazzone M, Dettori D, Leite De Oliveira R, et al. (2009) Heterozygous deficiency of PHD2 restores tumor oxygenation and inhibits metastasis via endothelial normalization. Cell 136: 839–851.

Pan Q, Chathery Y, Wu Y, et al. (2007) Neuropilin‐1 binds to VEGF121 and regulates endothelial cell migration and sprouting. The Journal of Biological Chemistry 282: 24049–24056.

Patel‐Hett S and D'amore PA (2011) Signal transduction in vasculogenesis and developmental angiogenesis. The International Journal of Developmental Biology 55: 353–363.

Pierce EA, Avery RL, Foley ED, Aiello LP and Smith LE (1995) Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization. Proceedings of the National Academy of Sciences of the United States of America 92: 905–909.

Pitulescu ME, Schmidt I, Benedito R and Adams RH (2010) Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice. Nature Protocols 5: 1518–1534.

Plein A, Fantin A, Denti L, Pollard JW and Ruhrberg C (2018) Erythro‐myeloid progenitors contribute endothelial cells to developing vasculature. Nature 562.

Raab S, Beck H, Gaumann A, et al. (2004) Impaired brain angiogenesis and neuronal apoptosis induced by conditional homozygous inactivation of vascular endothelial growth factor. Thrombosis and Haemostasis 91: 595–605.

Raimondi C, Fantin A, Lampropoulou A, et al. (2014) Imatinib inhibits VEGF‐independent angiogenesis by targeting neuropilin 1‐dependent ABL1 activation in endothelial cells. The Journal of Experimental Medicine 211: 1167–1183.

Risau W (1995) Differentiation of endothelium. The FASEB Journal 9: 926–933.

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 & Development 16: 2684–2698.

Ruhrberg C (2003) Growing and shaping the vascular tree: multiple roles for VEGF. BioEssays 25: 1052–1060.

Shalaby F, Rossant J, Yamaguchi TP, et al. (1995) Failure of blood‐island formation and vasculogenesis in Flk‐1‐deficient mice. Nature 376: 62–66.

Shen Q, Goderie SK, Jin L, et al. (2004) Endothelial cells stimulate self‐renewal and expand neurogenesis of neural stem cells. Science 304: 1338–1340.

Simons M, Alitalo K, Annex BH, et al. (2015) State‐of‐the‐art methods for evaluation of angiogenesis and tissue vascularization: a scientific statement from the American Heart Association. Circulation Research 116: e99–e132.

Smith LE, Wesolowski E, Mclellan A, et al. (1994) Oxygen‐induced retinopathy in the mouse. Investigative Ophthalmology & Visual Science 35: 101–111.

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

Soker S, Miao HQ, Nomi M, Takashima S and Klagsbrun M (2002) VEGF165 mediates formation of complexes containing VEGFR‐2 and neuropilin‐1 that enhance VEGF165‐receptor binding. Journal of Cellular Biochemistry 85: 357–368.

Suchting S, Freitas C, Le Noble F, et al. (2007) The Notch ligand Delta‐like 4 negatively regulates endothelial tip cell formation and vessel branching. Proceedings of the National Academy of Sciences of the United States of America 104: 3225–3230.

Tata M, Wall I, Joyce A, et al. (2016) Regulation of embryonic neurogenesis by germinal zone vasculature. Proceedings of the National Academy of Sciences of the United States of America 113: 13414–13419.

Valdembri D, Caswell PT, Anderson KI, et al. (2009) Neuropilin‐1/GIPC1 signaling regulates alpha5beta1 integrin traffic and function in endothelial cells. PLoS Biology 7: e25.

Zamir L, Singh R, Nathan E, et al. (2017) Nkx2.5 marks angioblasts that contribute to hemogenic endothelium of the endocardium and dorsal aorta. eLife 6.

Further Reading

Farrell AP (2001) Circulation in vertebrates. Encyclopedia of Life Sciences. DOI: 10.1038/npg.els.0001829.

Havrilak JA and Shannon JM (2015) Lung development. Encyclopedia of Life Sciences.

Lupo G, Caporarello N, Olivieri M, et al. (2017) Anti‐angiogenic therapy in cancer: downsides and new pivots for precision medicine. Frontiers in Pharmacology 07: 519.

Potente M, Gerhardt H and Carmeliet P (2011) Basic and therapeutic aspects of angiogenesis. Cell 146: 873–887.

Raimondi C, Brash JT, Fantin A and Ruhrberg C (2016) NRP1 function and targeting in neurovascular development and eye disease. Progress in Retinal and Eye Research 52: 64–83.

Scott A and Fruttiger M (2010) Oxygen‐induced retinopathy: a model for vascular pathology in the retina. Eye 24: 416–421.

Tata M and Ruhrberg C (2018) Cross‐talk between blood vessels and neural progenitors in the developing brain. Neuronal Signaling 2.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Bolton, Rebecca, Naylor, Kirsty, and Ruhrberg, Christiana(Aug 2019) Molecular and Cellular Mechanisms of Vascular Development. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0028519]