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Original Articles

Comparative Analysis of Vascular Endothelial Growth Factor Receptors on Retinal and Aortic Vascular Endothelial Cells

  1. Hagen Thieme,
  2. Lloyd Paul Aiello,
  3. Hitoshi Takagi,
  4. Napoleone Ferrara and
  5. George L King
  1. Research Division, Joslin Diabetes Center Boston, Massachusetts
  2. Beetham Eye Institute, Joslin Diabetes Center Boston, Massachusetts
  3. Harvard Medical School Boston, Massachusetts
  4. Department of Medicine, Brigham and Women's Hospital Boston, Massachusetts
  5. Genentech Inc San Francisco, California
  1. Address correspondence and reprint requests to Dr. Lloyd Paul Aiello, Beetham Eye Institute, Joslin Diabetes Center, One Joslin Place, Boston, MA 02215.
Diabetes 1995 Jan; 44(1): 98-103. https://doi.org/10.2337/diab.44.1.98
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Abstract

Ischemic eye disease often results in ocular neovascularization, presumably due to the elaboration of growth factors. Diabetic retinopathy is a classic example in which dramatic retinal neovascularization arises after ischemic retinal damage. The characterization of vascular endothelial growth factor (VEGF) as an angiogenic molecule whose expression is markedly induced by hypoxia makes it a promising candidate for mediating ischemic retinal neovascularization. Thus, we have characterized the structure, binding, and regulation of VEGF receptors in bovine retinal (BREC) and aortic endothelial cells (BAEC). VEGF stimulated a 2.1-fold increase in BREC number and DNA content at 0.6 nmol/l VEGF (P < 1 × 10−7). Scatchard binding analysis demonstrated specific high-affinity VEGF receptors on BREC with a Kd of 4.9 ± 0.6 × 10−11 mmol/l, similar to that observed for BAEC at 5.1 ± 0.4 × 10−11 mmol/l. BREC, however, possess 1.5 × 105 high-affinity receptors/cell, threefold more than BAEC (P < 0.003) and more than any cell type reported previously. 125I-VEGF affinity cross-linking revealed complexes at 220 and 170 kDa in BREC, but only a 220-kDa band of lesser intensity in BAEC. Cross-linking was displaceable in a dose-dependent manner by VEGF (P < 0.01) but not by other hormones. Hypoxia increased VEGF receptor number 50% in BREC without altering affinity. Antiphosphotyrosine immunoblotting showed VEGF-stimulated tyrosine autophosphorylation of VEGF receptor bands at 225 and 220 kDa and another band at 80 kDa within 1 min. These findings suggest that VEGF may mediate retinal vascular proliferation through large numbers of high-affinity receptors on retinal vascular endothelial cells and suggest that VEGF may be an important mediator of neovascularization induced by ischemic retinopathies such as diabetes.

  • Received April 26, 1994.
  • Revision received September 8, 1994.
  • Accepted September 8, 1994.
  • Copyright © 1995 by the American Diabetes Association

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January 1995, 44(1)
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Comparative Analysis of Vascular Endothelial Growth Factor Receptors on Retinal and Aortic Vascular Endothelial Cells
Hagen Thieme, Lloyd Paul Aiello, Hitoshi Takagi, Napoleone Ferrara, George L King
Diabetes Jan 1995, 44 (1) 98-103; DOI: 10.2337/diab.44.1.98

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Comparative Analysis of Vascular Endothelial Growth Factor Receptors on Retinal and Aortic Vascular Endothelial Cells
Hagen Thieme, Lloyd Paul Aiello, Hitoshi Takagi, Napoleone Ferrara, George L King
Diabetes Jan 1995, 44 (1) 98-103; DOI: 10.2337/diab.44.1.98
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More in this TOC Section

  • Intravitreal Triamcinolone Acetonide Inhibits Breakdown of the Blood-Retinal Barrier Through Differential Regulation of VEGF-A and Its Receptors in Early Diabetic Rat Retinas
  • Fibronectin Fragments Modulate Human Retinal Capillary Cell Proliferation and Migration
  • Overexpression of Glycogen Phosphorylase Increases GLUT4 Expression and Glucose Transport in Cultured Skeletal Human Muscle
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