May 2007
Volume 48, Issue 13
ARVO Annual Meeting Abstract  |   May 2007
Comparison of the Biological Activities of VEGF Isoforms
Author Affiliations & Notes
  • P. A. D'Amore
    Ophthalmology, Schepens Eye Res Inst / Harvard, Boston, Massachusetts
  • A. M. Goodwin
    Ophthalmology, Schepens Eye Research Institute, Boston, Massachusetts
  • S. K. Smith
    Children's Hospital, Boston, Massachusetts
  • E. Geretti
    Children's Hospital, Boston, Massachusetts
  • J. A. Nagy
    Beth Israel Deaconess Medical Center, Boston, Massachusetts
  • E. B. Finkelstein
    Ophthalmology, Schepens Eye Res Inst / Harvard, Boston, Massachusetts
  • H. F. Dvorak
    Beth Israel Deaconess Medical Center, Boston, Massachusetts
  • M. Saint-Geniez
    Ophthalmology, Schepens Eye Res Inst / Harvard, Boston, Massachusetts
  • Footnotes
    Commercial Relationships P.A. D'Amore, None; A.M. Goodwin, None; S.K. Smith, None; E. Geretti, None; J.A. Nagy, None; E.B. Finkelstein, None; H.F. Dvorak, None; M. Saint-Geniez, None.
  • Footnotes
    Support NIH Grant CA45548 and EY05318
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 1776. doi:
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    • Get Citation

      P. A. D'Amore, A. M. Goodwin, S. K. Smith, E. Geretti, J. A. Nagy, E. B. Finkelstein, H. F. Dvorak, M. Saint-Geniez; Comparison of the Biological Activities of VEGF Isoforms. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1776.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose:: VEGF-A in mice is produced as three alternatively spliced isoforms (VEGF-A120, VEGF-A164 and VEGF-A188) that differ in their binding affinity for heparan sulfate proteoglycan and thus in their extracellular localization following secretion. Transgenic mice that express single VEGF-A isoforms display distinct phenotypes. The goal of this work was to compare the biological activities of the three murine isoforms.

Methods:: Conditioned media (CM) containing the VEGF-A120, VEGF-A164 and VEGF-A188 produced by transient transfection of 293 cells with vectors expressing murine VEGF-A isoforms or media conditioned by 293 cells transfected with empty vectors were assayed for their effects on human umbilical vein endothelial cell (HUVEC) proliferation and tube formation. Levels of VEGFR-2 phosphorylation in response to the three isoforms were assessed using Phospho-VEGFR-2 (Tyr1175) and Total-VEGFR2 sandwich ELISA assays. Ability to bind neuropilin-1 (NP-1) was assessed in an in vitro binding assay. Effects on permeability were assessed in vivo using the Miles assay.

Results:: All three isoforms stimulated the proliferation of HUVEC to an equivalent extent, 2.7-fold increase over the control CM. Each isoform induced a dose-dependent increase in phosphorylation; however, the level of phosphorylation varied significantly: VEGF-A120> VEGF-A188> VEGF-A164. The induction of cord formation by HUVECs in response to the VEGF isoforms varied; VEGF-A188 was the most potent followed by VEGF-A164 and VEGF-A120. Binding of VEGF-A164, a known NP-1 ligand, to NP-1 was effectively competed by VEGF-A188 but not by VEGF-A120. Subcutaneous injection of each CM led to a dose dependent increase in permeability as detected by Evans blue extravasation, with no apparent difference among the isoforms.

Conclusions:: VEGF-A120, VEGF-A164 and VEGF-A188 have equal potency in stimulating endothelial proliferation and vascular permeability. However, the ability to induce VEGFR2 phosphorylation and formation of capillary-like structures in vitro differs and may be related to their abilities to be locally sequestered.

Keywords: vascular cells • growth factors/growth factor receptors • signal transduction 

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