Expression of VEGF Receptors on Endothelial Cells in Mouse Skeletal Muscle

Princess I. Imoukhuede, Aleksander S Popel

Research output: Contribution to journalArticle

Abstract

VEGFR surface localization plays a critical role in converting extracellular VEGF signaling towards angiogenic outcomes, and the quantitative characterization of these parameters is critical for advancing computational models; however the levels of these receptors on blood vessels is currently unknown. Therefore our aim is to quantitatively determine the VEGFR localization on endothelial cells from mouse hindlimb skeletal muscles. We contextualize this VEGFR quantification through comparison to VEGFR-levels on cells in vitro. Using quantitative fluorescence we measure and compare the levels of VEGFR1 and VEGFR2 on endothelial cells isolated from C57BL/6 and BALB/c gastrocnemius and tibialis anterior hindlimb muscles. Fluorescence measurements are calibrated using beads with known numbers of phycoerythrin molecules. The data show a 2-fold higher VEGFR1 surface localization relative to VEGFR2 with 2,000-3,700 VEGFR1/endothelial cell and 1,300-2,000 VEGFR2/endothelial cell. We determine that endothelial cells from the highly glycolytic muscle, tibialis anterior, contain 30% higher number of VEGFR1 surface receptors than gastrocnemius; BALB/c mice display ~17% higher number of VEGFR1 than C57BL/6. When we compare these results to mouse fibroblasts in vitro, we observe high levels of VEGFR1 (35,800/cell) and very low levels of VEGFR2 (700/cell), while in human endothelial cells in vitro, we observe that the balance of VEGFRs is inverted, with higher levels VEGFR2 (5,800/cell) and lower levels of VEGFR1 (1,800/cell). Our studies also reveal significant cell-to-cell heterogeneity in receptor expression, and the quantification of these dissimilarities ex vivo for the first time provides insight into the balance of anti-angiogenic or modulatory (VEGFR1) and pro-angiogenic (VEGFR2) signaling.

Original languageEnglish (US)
Article numbere44791
JournalPLoS One
Volume7
Issue number9
DOIs
StatePublished - Sep 12 2012

Fingerprint

vascular endothelial growth factor receptors
Vascular Endothelial Growth Factor Receptor
Endothelial cells
endothelial cells
Muscle
skeletal muscle
Skeletal Muscle
Endothelial Cells
mice
cells
Hindlimb
receptors
Fluorescence
Phycoerythrin
fluorescence
Blood vessels
Muscles
muscles
Fibroblasts
Vascular Endothelial Growth Factor A

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Expression of VEGF Receptors on Endothelial Cells in Mouse Skeletal Muscle. / Imoukhuede, Princess I.; Popel, Aleksander S.

In: PLoS One, Vol. 7, No. 9, e44791, 12.09.2012.

Research output: Contribution to journalArticle

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