Abstract
Vascular endothelial growth factor is an angiogenic factor in vivo and in vitro that plays a crucial role in the control of blood vessel development and in pathological angiogenesis. The vascularized extraembryonic membranes of the chick embryo include the area vasculosa and the chorioallantoic membrane. In this study, we investigated the expression of vascular endothelial growth factor and of its receptor-2, specifically expressed by the endothelial cells, in the chick area vasculosa at days 6, 10 and 14 of incubation. Our results indicate that, in all the three developmental stages examined, vascular endothelial growth factor is clearly expressed in the endodermal cells immediately adjacent to the mesodermal endothelial cells which, in turn, expressed vascular endothelial growth factor receptor-2. These observations suggest that during the development of the vascular system, endodermal cells, expressing vascular endothelial growth factor, initiate angiogenesis by stimulating directly mesodermal cells, which express vascular endothelial growth factor receptor-2. Moreover, our data demonstrate that vascular endothelial growth factor receptor-2 expression is also maintained by endothelial cells in the later stages of development, until day 14 of incubation. In accord with other literature data, this suggests that vascular endothelial growth factor is required not only for proliferation, but also for the survival of endothelial cells.
Similar content being viewed by others
References
Augustine JM (1970) Expansion of the area vasculosa of the chick after removal of the ectoderm. J Embryol Exp Morphol 24: 95–108.
Boocock CA, Charnock-Jones DS, Sharkey AM, Mc Laren J, Barker PJ, Wright KA, Twentyman PR, Smith SK (1995) Expression of vascular endothelial growth factor and of its receptors flt-1 and KDR in ovarian carcinoma. J Natl Cancer Inst 87: 506–516.
Breier G, Albrecht U, Sterrer S, Risau W (1992) Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. Development 114: 521–532.
Breier G, Clauss M, Risau W (1995) Coordinate expression of vascular endothelial growth factor receptor-1 (Flt-1) and its ligand suggests a paracrine regulation of murine vascular development. Dev Dyn 204: 228–239.
Brown LF, Detmar M, Tognazzi K, Abu-Jawdeh G, Iruela-Arispe ML (1997) Uterine smooth muscle cells express functional receptors (flt-1 and KDR) for vascular permeability factor/vascular endothelial growth factor. Lab Invest 76: 245–255.
Carlevaro MF, Cermelli S, Cancedda R, Descalzi Cancedda F (2000) Vascular endothelial growth factor (VEGF) in cartilage neovascularization and chondrocyte differentiation: auto-paracrine role during endochondral bone formation. J Cell Sci 113: 59–69.
Carmeliet P, Ferreira V, Breier G, Pollefeyt S, Kiekens L, Gertsenstein M, Fahrig M, Vandenhoeck A, Harpal K, Eberhardt C, Declercq C, Pawling J, Moons L, Collen D, Risau W, Nagy A (1996) Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380: 435–439.
Dickson MC, Martin JS, Cousins FM, Kulkarni AB, Karlsson S, Akhurst RJ (1995) Defective haematopoiesis and vasculogenesis in transforming growth factor-beta 1 knock out mice. Development 21: 1845–1854.
Dumont DJ, Fong GH, Puri MC, Gradwohl G, Alitalo K, Breitman ML (1995) Vascularization of the mouse embryo: a study of flk-1, tek, tie, and vascular endothelial growth factor expression during development. Dev Dyn 203: 80–92.
Dumont DJ, Jussila L, Taipale J, Mustonen T, Pajusola K, Breitman M, Alitalo K (1998) Failure of cardiovascular development in embryos deficient of the lymphatic vascular endothelial growth factor-C receptor, VEGFR-3. Science 282: 946–949.
Dvorak HF, Brown LF, Detmar M, Dvorak AM (1995) Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol 146: 1029–1039.
Eichmann A, Marcelle C, Breant C, Le Douarin N (1993) Two molecules related to the VEGF receptor are expressed in early endothelial cells during embryonic development. Mech Dev 42: 33–48.
Ferrara N, Carver-Moore K, Chen H, Dowd M, Lu L, O'shea KS, Powell-Braxton L, Hillan KJ, Moore MW (1996) Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene.Nature 380: 439–442.
Flamme I, Risau W (1992) Induction of vasculogenesis and hematopoiesis in vitro. Development 116: 435–439.
Flamme I, Breier G, Risau W (1995) Vascular endothelial growth factor (VEGF) and VEGF receptor-2 (flk-1) are expressed during vasculogenesis and vascular differentiation in quail embryo. Dev Biol 169: 699–712.
Fong GH, Rossant, Gertsenstein M, Breitman ML (1995) Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376: 66–70.
Gerber HP, Hillan KJ, Ryan AM, Kowalski J, Keller GA, Rangell L, Wright BD, Radtre F, Aguet M, Ferrara N (1999) VEGF is required for growth and survival in neonatal mice. Development 126: 1149–1159.
Leveen P, Pekny M, Gebre-Medhin S, Swolin B, Larsson E, Betsholtz C (1994) Mice deficient for PDGF B show renal, cardiovascular and hematological abnormalities. Genes Dev 8: 1875–1887.
Mayer BW, Packard JR (1978) A study of the expansion of the chick area vasculosa. Dev Biol 85: 429–450.
Oshima M, Oshima H, Taketo MM(1996) TGF-beta receptor type II defi-ciency results in defects of yolk sac hematopoiesis and vasculogenesis. Dev Biol 179: 297–302.
Pardanuad L, Dieterlen-Lievre F (1999). Manipulation of the angiopoietic/hematopoietic commitment in the avian embryo. Development 126: 617–627.
Risau W (1997) Mechanism of angiogenesis. Nature 386: 671–674.
Sato TN, Tozawa Y, Deutsch U, Wolburg-Buchholz K, Fujiwara Y, Gendron-Maguire M, Gridley T, Wolburg H, Risau W, Quin Y (1995) Distinct roles of the receptor tyrosine kinases Tie-1 and Tie-2 in blood vessel formation. Nature 376: 70–74.
Serra JA (1946) Histochemical tests for protein and amino acids. The characterization of basic proteins. Stain Technol 21: 5–18.
Shalaby F, Rossant J, Yamaguchi TP, Gertsenstein M, Wu XF, Breitman ML, Schuh AC (1995) Failure of blood island formation and vasculogenesis in Flk-1-deficient mice. Nature 376: 62–66.
Shalaby F, Ho J, Fisher KD, Schuh AC, Schwartz L, Bernstein A, Rossant J (1997) A requirement for Flk-1 in primitive and definitive hematopoiesis and vasculogenesis. Cell 89: 981–990.
Soriano P (1994). Abnormal kidney development and hematological disorders in PDGF B-receptor mutant mice. Genes Dev 8: 1888–1896.
Spiers V, Atkin SL (1999). Production of VEGF and expression of the VEGF receptors flt-1 and KDR in primary cultures of epithelial cells and stromal cells derived from breast tumours. Br J Cancer 80: 893–903.
Suri C, Jones PF, Patan S, Bartunkova S, Maisonpierre PC, Davis S, Sato TN, Yancopouolos GD (1996) Requisite role of agiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell 87: 1171–1180.
Wang HU, Chen Z, Anderson DJ (1988) Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by ephrin-2 and its receptor eph-B4. Cell 93: 741–753.
Wilt FH (1964) Erythropoiesis in the chick embryo: the role of endoderm. Science 147: 1588–1590.
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: 489–498.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nico, B., Vacca, A., De Giorgis, M. et al. Vascular Endothelial Growth Factor and Vascular Endothelial Growth Factor Receptor-2 Expression in the Chick Embryo Area Vasculosa. Histochem J 33, 283–286 (2001). https://doi.org/10.1023/A:1017977007479
Issue Date:
DOI: https://doi.org/10.1023/A:1017977007479