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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1992 Jan;89(1):244–253. doi: 10.1172/JCI115568

Binding sites for vascular endothelial growth factor are localized on endothelial cells in adult rat tissues.

L B Jakeman 1, J Winer 1, G L Bennett 1, C A Altar 1, N Ferrara 1
PMCID: PMC442842  PMID: 1729274

Abstract

Vascular endothelial growth factor (VEGF) is a secreted heparin-binding mitogen; its growth-promoting activity is limited to vascular endothelial cells in vitro and VEGF also stimulates angiogenesis in vivo. To identify target cells for VEGF and investigate the potential physiological role of this factor, iodinated recombinant human VEGF (125I-rhVEGF) was used for in vitro ligand autoradiography on tissue sections from adult rats. 125I-rhVEGF exhibited saturable, displaceable binding to a single class of sites with high affinity and low capacity in all tissues and organs examined. Colocalization of 125I-rhVEGF binding with Factor VIII-like immunoreactivity demonstrated binding sites associated with vascular endothelial cells of both fenestrated and nonfenestrated microvessels and the endothelium of large vessels, while no displaceable binding was evident on nonendothelial cells. Specific binding was associated with quiescent as well as proliferating vessels. These findings support the hypothesis that VEGF plays a specific role in both the maintenance and in the induction of growth of vascular endothelial cells.

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  1. Aggarwal B. B., Eessalu T. E., Hass P. E. Characterization of receptors for human tumour necrosis factor and their regulation by gamma-interferon. Nature. 1985 Dec 19;318(6047):665–667. doi: 10.1038/318665a0. [DOI] [PubMed] [Google Scholar]
  2. Altar C. A., Walter R. J., Jr, Neve K. A., Marshall J. F. Computer-assisted video analysis of [3H]spiroperidol binding autoradiographs. J Neurosci Methods. 1984 Mar;10(3):173–188. doi: 10.1016/0165-0270(84)90054-2. [DOI] [PubMed] [Google Scholar]
  3. Betsholtz C., Rorsman F., Westermark B., Ostman A., Heldin C. H. Analogous alternative splicing. Nature. 1990 Mar 22;344(6264):299–299. doi: 10.1038/344299a0. [DOI] [PubMed] [Google Scholar]
  4. Clauss M., Gerlach M., Gerlach H., Brett J., Wang F., Familletti P. C., Pan Y. C., Olander J. V., Connolly D. T., Stern D. Vascular permeability factor: a tumor-derived polypeptide that induces endothelial cell and monocyte procoagulant activity, and promotes monocyte migration. J Exp Med. 1990 Dec 1;172(6):1535–1545. doi: 10.1084/jem.172.6.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Conn G., Bayne M. L., Soderman D. D., Kwok P. W., Sullivan K. A., Palisi T. M., Hope D. A., Thomas K. A. Amino acid and cDNA sequences of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2628–2632. doi: 10.1073/pnas.87.7.2628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Connolly D. T., Knight M. B., Harakas N. K., Wittwer A. J., Feder J. Determination of the number of endothelial cells in culture using an acid phosphatase assay. Anal Biochem. 1986 Jan;152(1):136–140. doi: 10.1016/0003-2697(86)90131-4. [DOI] [PubMed] [Google Scholar]
  7. Cuatrecasas P., Hollenberg M. D. Membrane receptors and hormone action. Adv Protein Chem. 1976;30:251–451. doi: 10.1016/s0065-3233(08)60481-7. [DOI] [PubMed] [Google Scholar]
  8. Ferrara N., Henzel W. J. Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells. Biochem Biophys Res Commun. 1989 Jun 15;161(2):851–858. doi: 10.1016/0006-291x(89)92678-8. [DOI] [PubMed] [Google Scholar]
  9. Folkman J., Klagsbrun M. Angiogenic factors. Science. 1987 Jan 23;235(4787):442–447. doi: 10.1126/science.2432664. [DOI] [PubMed] [Google Scholar]
  10. Gimenez-Gallego G., Rodkey J., Bennett C., Rios-Candelore M., DiSalvo J., Thomas K. Brain-derived acidic fibroblast growth factor: complete amino acid sequence and homologies. Science. 1985 Dec 20;230(4732):1385–1388. doi: 10.1126/science.4071057. [DOI] [PubMed] [Google Scholar]
  11. Gospodarowicz D., Ferrara N., Schweigerer L., Neufeld G. Structural characterization and biological functions of fibroblast growth factor. Endocr Rev. 1987 May;8(2):95–114. doi: 10.1210/edrv-8-2-95. [DOI] [PubMed] [Google Scholar]
  12. Gospodarowicz D., Massoglia S., Cheng J., Fujii D. K. Effect of fibroblast growth factor and lipoproteins on the proliferation of endothelial cells derived from bovine adrenal cortex, brain cortex, and corpus luteum capillaries. J Cell Physiol. 1986 Apr;127(1):121–136. doi: 10.1002/jcp.1041270116. [DOI] [PubMed] [Google Scholar]
  13. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  14. Ishikawa F., Miyazono K., Hellman U., Drexler H., Wernstedt C., Hagiwara K., Usuki K., Takaku F., Risau W., Heldin C. H. Identification of angiogenic activity and the cloning and expression of platelet-derived endothelial cell growth factor. Nature. 1989 Apr 13;338(6216):557–562. doi: 10.1038/338557a0. [DOI] [PubMed] [Google Scholar]
  15. Keck P. J., Hauser S. D., Krivi G., Sanzo K., Warren T., Feder J., Connolly D. T. Vascular permeability factor, an endothelial cell mitogen related to PDGF. Science. 1989 Dec 8;246(4935):1309–1312. doi: 10.1126/science.2479987. [DOI] [PubMed] [Google Scholar]
  16. LEPESCHKIN E. On the relation between the site of valvular involvement in endocarditis and the blood pressure resting on the valve. Am J Med Sci. 1952 Sep;224(3):318–319. doi: 10.1097/00000441-195209000-00011. [DOI] [PubMed] [Google Scholar]
  17. Leung D. W., Cachianes G., Kuang W. J., Goeddel D. V., Ferrara N. Vascular endothelial growth factor is a secreted angiogenic mitogen. Science. 1989 Dec 8;246(4935):1306–1309. doi: 10.1126/science.2479986. [DOI] [PubMed] [Google Scholar]
  18. Levy A. P., Tamargo R., Brem H., Nathans D. An endothelial cell growth factor from the mouse neuroblastoma cell line NB41. Growth Factors. 1989;2(1):9–19. doi: 10.3109/08977198909069077. [DOI] [PubMed] [Google Scholar]
  19. Madri J. A., Bell L., Marx M., Merwin J. R., Basson C., Prinz C. Effects of soluble factors and extracellular matrix components on vascular cell behavior in vitro and in vivo: models of de-endothelialization and repair. J Cell Biochem. 1991 Feb;45(2):123–130. doi: 10.1002/jcb.240450202. [DOI] [PubMed] [Google Scholar]
  20. Moyer C. F., Dennis P. A., Majno G., Joris I. Venular endothelium in vitro: isolation and characterization. In Vitro Cell Dev Biol. 1988 Apr;24(4):359–368. doi: 10.1007/BF02628839. [DOI] [PubMed] [Google Scholar]
  21. Mukai K., Rosai J., Burgdorf W. H. Localization of factor VIII-related antigen in vascular endothelial cells using an immunoperoxidase method. Am J Surg Pathol. 1980 Jun;4(3):273–276. doi: 10.1097/00000478-198006000-00008. [DOI] [PubMed] [Google Scholar]
  22. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  23. Phillips H. S., Hains J., Leung D. W., Ferrara N. Vascular endothelial growth factor is expressed in rat corpus luteum. Endocrinology. 1990 Aug;127(2):965–967. doi: 10.1210/endo-127-2-965. [DOI] [PubMed] [Google Scholar]
  24. Plouet J., Moukadiri H. Specific binding of vasculotropin to bovine brain capillary endothelial cells. Biochimie. 1990 Jan;72(1):51–55. doi: 10.1016/0300-9084(90)90172-d. [DOI] [PubMed] [Google Scholar]
  25. Plouët J., Moukadiri H. Characterization of the receptor to vasculotropin on bovine adrenal cortex-derived capillary endothelial cells. J Biol Chem. 1990 Dec 25;265(36):22071–22074. [PubMed] [Google Scholar]
  26. Plouët J., Schilling J., Gospodarowicz D. Isolation and characterization of a newly identified endothelial cell mitogen produced by AtT-20 cells. EMBO J. 1989 Dec 1;8(12):3801–3806. doi: 10.1002/j.1460-2075.1989.tb08557.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reidy M. A., Schwartz S. M. Endothelial regeneration. III. Time course of intimal changes after small defined injury to rat aortic endothelium. Lab Invest. 1981 Apr;44(4):301–308. [PubMed] [Google Scholar]
  28. Rodbard D., Lewald J. E. Computer analysis of radioligand assay and radioimmunoassay data. Acta Endocrinol Suppl (Copenh) 1970;147:79–103. doi: 10.1530/acta.0.065s079. [DOI] [PubMed] [Google Scholar]
  29. Rosenthal R. A., Megyesi J. F., Henzel W. J., Ferrara N., Folkman J. Conditioned medium from mouse sarcoma 180 cells contains vascular endothelial growth factor. Growth Factors. 1990;4(1):53–59. doi: 10.3109/08977199009011010. [DOI] [PubMed] [Google Scholar]
  30. Senger D. R., Galli S. J., Dvorak A. M., Perruzzi C. A., Harvey V. S., Dvorak H. F. Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science. 1983 Feb 25;219(4587):983–985. doi: 10.1126/science.6823562. [DOI] [PubMed] [Google Scholar]
  31. Unnerstall J. R., Niehoff D. L., Kuhar M. J., Palacios J. M. Quantitative receptor autoradiography using [3H]ultrofilm: application to multiple benzodiazepine receptors. J Neurosci Methods. 1982 Jul;6(1-2):59–73. doi: 10.1016/0165-0270(82)90016-4. [DOI] [PubMed] [Google Scholar]
  32. Vaisman N., Gospodarowicz D., Neufeld G. Characterization of the receptors for vascular endothelial growth factor. J Biol Chem. 1990 Nov 15;265(32):19461–19466. [PubMed] [Google Scholar]
  33. Vlodavsky I., Folkman J., Sullivan R., Fridman R., Ishai-Michaeli R., Sasse J., Klagsbrun M. Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2292–2296. doi: 10.1073/pnas.84.8.2292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Walter P., Blobel G. Translocation of proteins across the endoplasmic reticulum III. Signal recognition protein (SRP) causes signal sequence-dependent and site-specific arrest of chain elongation that is released by microsomal membranes. J Cell Biol. 1981 Nov;91(2 Pt 1):557–561. doi: 10.1083/jcb.91.2.557. [DOI] [PMC free article] [PubMed] [Google Scholar]

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