Novel role of ARF6 in vascular endothelial growth factor-induced signaling and angiogenesis

Satoshi Ikeda, Masuko Ushio-Fukai, Lian Zuo, Taiki Tojo, Sergey Dikalov, Nikolay A. Patrushev, R. Wayne Alexander

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

Vascular endothelial growth factor (VEGF) stimulates endothelial cell (EC) migration and proliferation primarily through the VEGF receptor-2 (VEGFR2). We have shown that VEGF stimulates a Rac1-dependent NAD(P)H oxidase to produce reactive oxygen species (ROS) that are involved in VEGFR2 autophosphorylation and angiogenic-related responses in ECs. The small GTPase ARF6 is involved in membrane trafficking and cell motility; however, its roles in VEGF signaling and physiological responses in ECs are unknown. In this study, we show that overexpression of dominant-negative ARF6 [ARF6(T27N)] almost completely inhibits VEGF-induced Rac1 activation, ROS production, and VEGFR2 autophosphorylation in ECs. Fractionation of caveolae/lipid raft membranes demonstrates that ARF6, Rac1, and VEGFR2 are localized in caveolin-enriched fractions basally. VEGF stimulation results in the release of VEGFR2 from caveolae/lipid rafts and caveolin-1 without affecting localization of ARF6, Rac1, or caveolin-1 in these fractions. The egress of VEGFR2 from caveolae/lipid rafts is contemporaneous with the tyrosine phosphorylation of caveolin-1 (Tyr14) and VEGFR2 and with their association with each other. ARF6(T27N) significantly inhibits both VEGF-induced responses. Immunofluorescence studies show that activated VEGFR2 and phosphocaveolin colocalize at focal complexes/adhesions after VEGF stimulation. Both overexpression of ARF6(T27N) and mutant caveolin-1(Y14F), which cannot be phosphorylated, block VEGF-stimulated EC migration and proliferation. Moreover, ARF6 expression is markedly upregulated in association with an increase in capillary density in a mouse hindlimb ischemia model of angiogenesis. Thus, ARF6 is involved in the temporal-spatial organization of caveolae/lipid rafts- and ROS-dependent VEGF signaling in ECs as well as in angiogenesis in vivo.

Original languageEnglish (US)
Pages (from-to)467-475
Number of pages9
JournalCirculation research
Volume96
Issue number4
DOIs
StatePublished - Mar 23 2005
Externally publishedYes

Fingerprint

Vascular Endothelial Growth Factor Receptor
Vascular Endothelial Growth Factor A
Caveolin 1
Caveolae
Cell Movement
Reactive Oxygen Species
Lipids
Endothelial Cells
Cell Proliferation
Caveolins
Vascular Endothelial Growth Factor Receptor-2
Focal Adhesions
Monomeric GTP-Binding Proteins
NADPH Oxidase
Hindlimb
Membrane Lipids
Fluorescent Antibody Technique
Tyrosine
Ischemia
Phosphorylation

Keywords

  • ARF6
  • Angiogenesis
  • Caveolin
  • Endothelial cell
  • Vascular endothelial growth factor

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Novel role of ARF6 in vascular endothelial growth factor-induced signaling and angiogenesis. / Ikeda, Satoshi; Ushio-Fukai, Masuko; Zuo, Lian; Tojo, Taiki; Dikalov, Sergey; Patrushev, Nikolay A.; Alexander, R. Wayne.

In: Circulation research, Vol. 96, No. 4, 23.03.2005, p. 467-475.

Research output: Contribution to journalArticle

Ikeda, Satoshi ; Ushio-Fukai, Masuko ; Zuo, Lian ; Tojo, Taiki ; Dikalov, Sergey ; Patrushev, Nikolay A. ; Alexander, R. Wayne. / Novel role of ARF6 in vascular endothelial growth factor-induced signaling and angiogenesis. In: Circulation research. 2005 ; Vol. 96, No. 4. pp. 467-475.
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