Skip to main content
SpringerLink
Log in

The effects of growth factors on the day 13 chorioallantoic membrane (CAM): a study of VEGF165 and PDGF-BB

  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Summary

The in vivo effects of two growth factors, VEGF165 and PDGF-BB, were studied in the chick chorioallantoic membrane (CAM). The factors were airdried on Thermanox discs and the inverted discs were placed on the day-13 CAM for a period of 3 days. The specimens were then fixed, examined under a stereomicroscope and processed for semi- and ultrathin sectioning. VEGF165 induces marked vascular growth. Many new blood vessels emerge from the precapillary arterioles, and a brush-like formation of vessels can be seen in this region. In the venous part of the vascular system, the formation of sinusoidal or lacunar vessels can be seen. Edema does not develop. PDGF-BB induces thickening of the CAM due to extracellular-matrix production and the proliferation or immigration of fibrocytes. These lie just beneath the ectodermal epithelium and are oriented parallel to it. Out of the four factors we have already studied (PDGF-BB, VEGF165, Angiogenin, bFGF), only VEGF165 specifically induces the growth of blood vessels.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Antoniades HN, Galanopoulos T, Neville-Golden J, Kiritsy CP, Lynch SE (1991) Injury induces in vivo expression of platelet-derived growth factor (PDGF) and PDGF receptor mRNAs in skin epithelial cells and PDGF mRNA in connective tissue fibroblasts. Proc Natl Acad Sci USA 88:565–569

    Google Scholar 

  • Auerbach R, Auerbach W, Polakowsky J (1991) Assays for angiogenesis: a review. Pharmacol Ther 51:1–11

    Google Scholar 

  • Ausprunk DH, Folkman J (1977) Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis. Microvasc Res 14:53–65

    Google Scholar 

  • Ausprunk DH, Knighton DR, Folkman J (1974) Differentiation of the vascular endothelium in the chick chorioallantois: a structural and autoradiographic study

  • Bazan HEP, Reddy STK, Lin N (1991) Platelet-activating factor (PAF) accumulation correlates with injury in the cornea. Exp Eye Res 52:481–491

    Google Scholar 

  • Beckmann MP, Betsholtz C, Heldin CH, Westermark B, Di Marco E, Di Fiore PP, Robbins KC, Aaronson SA (1988) Comparison of biological properties and transforming potential of human PDGF-A and PDGF-B chains. Science 241:1346–1349

    Google Scholar 

  • Beitz JG, In-Soon K, Calabresi P, Frackelton Jr R (1991) Human microvascular endothelial cells express receptors for platelet-derived growth factor. Proc Natl Acad Sci USA 88:2021–2025

    Google Scholar 

  • Berta A, Holly FJ, Tözser J, Holly TF (1991) Tear plasminogen activators-indicators of epithelial cell destruction. The effect of scraping, n-heptanol debridement, and alkali burn of the cornea on the plasminogen activator activity of rabbit tears. Int Ophthalmol 15:363–369

    Google Scholar 

  • Bicknell R, Harris AL (1991) Novel growth regulatory factors and tumour angiogenesis. Eur J Cancer 27:781–785

    Google Scholar 

  • Bikfalvi A, Sauzeau C, Moukadiri H, Maclouf J, Busso N, Bryckaert M, Plouët J, Tobelem G (1991) Interaction of vasculotropin/vascular endothelial cell growth factor with human umbilical vein endothelial cells: Binding, internalization, degradation, and biological effects. J Cell Physiol 149:50–59

    Google Scholar 

  • 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

    Google Scholar 

  • Conn G, Bayne ML, Soderman DD, Kwok PW, Sullivan KA, Palisi TM, Hope DA, Thomas KA (1990) Amino acid and cDNA sequences of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor. Proc Natl Acad Sci USA 87:2628–2632

    Google Scholar 

  • Connolly DT, Heuvelman D, Nelson R, Olander JV, Eppler BL, Delfino JJ, Siegel NR, Leimgruber RM, Feder J (1989) Tumor vascular permeability factor stimulates endothelial cell growth and angiogenesis. J Clin Invest 84:1470–1478

    CAS  PubMed  Google Scholar 

  • Dvorak HF, Sioussat TM, Brown LF, Berse B, Nagy JA, Sotrel A, Manseau EJ, Van De Water L, Senger DR (1991) Distribution of vascular permeability factor (vascular endothelial growth factor) in tumours: concentration in tumor blood vessels. J Exp Med 174:1275–1278

    Google Scholar 

  • Ferrara N, Henzel WJ (1989) Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells. Biochem Biophys Res Commun 161:851–858

    CAS  PubMed  Google Scholar 

  • Ferrara N, Keith AH, Jakeman LB, Winer J, Leung DW (1991) The vascular endothelial growth factor family of polypeptides. J Cell Biochem 47:211–218

    Google Scholar 

  • Folkman J (1985) Tumor angiogenesis. Adv Canc Res 43:175–203

    Google Scholar 

  • Gilardetti RS, Chaibi MS, Stroumza J, Williams SR, Antoniades HN, Carnes DC, Graves DT (1991) High-affinity binding of PDGF-AA and PDGF-BB to normal human osteoblastic cells and modulation by interleukin-1. Am J Physiol 261 (Cell Physiol 30):C980–985

    Google Scholar 

  • Gospodarowicz D, Abraham JA, Schilling J (1989) Isolation and characterization of a vascular endothelial cell mitogen produced by pituitary-derived folliculo stellate cells. Proc Natl Acad Sci USA 86:7311–7315

    Google Scholar 

  • Grotendorst GR, Igarashi A, Larson R, Soma Y, Charette M (1991) Differential binding, biological and biochemical actions of recombinant PDGF AA, AB and BB molecules on connective tissue cells. J Cell Physiol 149:235–243

    Google Scholar 

  • Heldin CH, Backstrom G, Ostman A, Hammacher A, Ronnstrand L, Rubin K, Nister M, Westermark B (1988) Binding of different dimeric forms of PDGF to human fibroblasts: evidence for two separate receptor types. EMBO J 7:1387–1393

    Google Scholar 

  • Holmgren L, Glaser A, Pfeifer-Ohlsson S, Ohlsson R (1991) Angiogenesis during human extraembryonic development involves the spatiotemporal control of PDGF ligand and receptor gene expression. Development 113:749–754

    Google Scholar 

  • Jakeman LB, Winer J, Bennet GL, Altar CA, Ferrara N (1992) Binding sites for vascular endothelial growth factor are localized on endothelial cells in adult rat tissues. J Clin Invest 89:244–253

    Google Scholar 

  • Jakob W, Jentzsch KD, Mauersberger B, Heder G (1978) The chick embryo choriallantoic membrane as a bioassay for angiogenesis factors: reactions induced by carrier materials. Exp Pathol 15:241–249

    Google Scholar 

  • Johnson A, Heldin CH, Westermark B, Wasteson A (1982) Platelet-derived growth factor: identification of constituent polypeptide chains. Biochem Biophys Res Commun 104:66–74

    Google Scholar 

  • Keck PJ, Hauser SD, Krivi G, Sanzo K, Warren T, Feder J, Connolly DT (1989) Vascular permeability factor, an endothelial cell mitogen related to PDGF. Science 246:1309–1312

    Google Scholar 

  • Leung DW, Cachianes G, Kuang WJ, Goeddel DV, Ferrara N (1989) Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 246:1306–1309

    CAS  PubMed  Google Scholar 

  • Ogawa Y, Ksander GA, Pratt BM, Sawamura SJ, Ziman JM, Gerhardt CO, Avis PD, Murray MJ, McPherson JM (1991) Differences in the biological activities of transforming growth factor-β and platelet-derived growth factor in vivo. Growth Factors 5:57–68

    Google Scholar 

  • Plouët J, Moukadiri H (1990) Characterization of the receptor to vasculotropin on bovine adrenal cortex-derived capillary endothelial cells. J Biol Chem 265:22071–22074

    Google Scholar 

  • Plouët J, Schilling J, Gospodarowicz D (1989) Isolation and characterization of a newly identified endothelial cell mitogen produced by AfT-20 cells. EMBO J 8:3801–3806

    Google Scholar 

  • Sakai J, Hung J, Zhu G, Katakami C, Boyce S, Kao WWY (1991) Collagen metabolism during healing of lacerated rabbit corneas. Exp Eye Res 52:237–244

    Google Scholar 

  • Senger DR, Galli SJ, Dvorak AM, Perruzzi CA, Harvey VS, Dvorak HF (1983) Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science 219:983–985

    Google Scholar 

  • Silver BJ, Jaffer FE, Abboud HE (1988) Platelet-derived growth factor synthesis in mesangial cells: induction by multiple peptide mitogens. Proc Natl Acad Sci USA 86:1056–1060

    Google Scholar 

  • Spanel-Borowski K, Schnapper U, Heymer B (1988) The chick chorioallantoic membrane assay in the assessment of angiogenic factors. Biomed Res 9(4): 253–261

    Google Scholar 

  • Tischer E, Gospodarowicz D, Mitchell R, Silva M, Schilling J, Lau K, Crisp T, Fiddes JC, Abraham JA (1989) Vascular endothelial growth factor: a new member of the platelet-derived growth factor gene family. Biochem Biophys Res Commun 165:1198–1206

    Google Scholar 

  • Vaisman N, Gospodarowicz D, Neufeld G (1990) Characterization of the receptors for vascular endothelial growth factor. J Biol Chem 265:19461–19466

    Google Scholar 

  • Vallee BL, Riordan JF, Lobb RR, Higachi N, Fett JW, Crossley G, Bühler R, Budzik G, Breddam K, Bethune JL, Alderman EM (1985) Tumor-derived angiogenesis factors from rat Walker 256 carcinoma: an experimental investigation and review. Experientia 41:1–42

    Google Scholar 

  • Wilting J, Christ B, Bokeloh M (1991) A modified chorioallantoic membrane (CAM) assay for qualitative and quantitative study of growth factors. Studies on the effect of carriers, PBS, angiogenin and bFGF. Anat Embryol 183:259–271

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Anatomisches Institut II der Albert-Ludwigs-Universität, Albertstrasse 17, W-7800, Freiburg, Federal Republic of Germany

    Jörg Wilting & Bodo Christ

  2. Institut für Molekulare Zellbiologie der Albert-Ludwigs-Universität, c/o Gödecke AG, Mooswaldallee 1-9, W-7800, Freiburg, Federal Republic of Germany

    Herbert A. Weich

Authors
  1. Jörg Wilting
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bodo Christ
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Herbert A. Weich
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wilting, J., Christ, B. & Weich, H.A. The effects of growth factors on the day 13 chorioallantoic membrane (CAM): a study of VEGF165 and PDGF-BB. Anat Embryol 186, 251–257 (1992). https://doi.org/10.1007/BF00174147

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00174147

Key words

Search

Navigation