Growth factor- and heparin-dependent regulation of constitutive and agonist-mediated human endothelial barrier function

Moy, Alan B.; Blackwell, Ken; Wu, Mack H.; Granger, Harris J.

doi:10.1152/ajpheart.00185.2006

Cited by 5 publications

(7 citation statements)

References 31 publications

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“…We chose TER measurement to assess barrier function because it has been previously established as a reliable index of permeability of vascular endothelial monolayers 9 , which is verified by the fact that the response we observed to VEGF-A reflects previously published findings 8,[14][15][16]31 . TER also has the advantage insofar that rapid, dynamic changes can be measured, whereas solute flux studies are generally less sensitive because they are limited to determining average permeability coefficients over longer time periods required to accumulate enough tracer to accurately measure flux.…”

Section: Discussionsupporting

confidence: 54%

“…We first tested the ability of the endothelial monolayers to respond to VEGF-A (Fig. 2), which increases endothelial permeability both in vivo and in cultured endothelial cell models 8,[14][15][16]31 . In HUVEC monolayers, VEGF-A caused a time-dependent decrease in TER, indicating an opening of the endothelial barrier, followed by a recovery characterized by elevated TER.…”

Section: Resultsmentioning

confidence: 99%

“…Cells were treated with the indicated concentrations of VEGF-A, VEGF-C, VEGF-C156S, or MAZ51. The optimal concentrations used were based upon our dose-response study and work reported by others 8,[27][28][29][30][31] . Endothelial barrier function is presented as a fractional change in TER.…”

Section: Cell Culture and Ter Measurementmentioning

confidence: 99%

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VEGF-C Alters Barrier Function of Cultured Lymphatic Endothelial Cells Through a VEGFR-3-Dependent Mechanism

Breslin

Yuan

2007

Lymphatic Research and Biology

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Background-The lymphatic endothelium is an important semi-permeable barrier separating lymph from the interstitial space. However, there is currently a limited understanding of the lymphatic endothelial barrier and the mechanisms of lymph formation. The objectives of this study were to investigate the potential active role of lymphatic endothelial cells in barrier regulation, and to test whether the endothelial cell agonists VEGF-A and VEGF-C can alter lymphatic endothelial barrier function.

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Section: Discussionsupporting

confidence: 54%

Section: Resultsmentioning

confidence: 99%

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VEGF-C Alters Barrier Function of Cultured Lymphatic Endothelial Cells Through a VEGFR-3-Dependent Mechanism

Breslin

Yuan

2007

Lymphatic Research and Biology

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“…We hypothesized, therefore, that any agent causing an abrupt increase in permeability would likewise trigger an upstream increase in shear rate and subsequent dilation. Figure 8 shows the remote response with histamine and thrombin, each well documented to increase local permeability [33,34]. Figure 8 shows that each agent caused an abrupt increase in upstream velocity and shear rate, which was followed by a remote vasodilation.…”

Section: Resultsmentioning

confidence: 99%

Downstream Exposure to Growth Factors Causes Elevated Velocity and Dilation in Arteriolar Networks

2010

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Our goal was to characterize changes in flow and diameter with vascular endothelial cell growth factor A (VEGF-A) and fibroblast growth factor 2 (FGF2). Observations were made in arteriolar networks of the cheek pouch tissue in anesthetized hamsters (pentobarbital 70 mg/kg, i.p., n = 45). Local and remote dilation responses to micropipette-applied VEGF or FGF2 yielded similar EC₅₀ values. The role of gap junctions in the remote response was tested by applying sucrose, halothane or 18αGA to the feed arteriole midway between the remote stimulation and upstream observation sites; all remote dilation to FGF2 was prevented, while only the early dilation to VEGF was blocked. The remote dilation to VEGF displayed a second rheologic mechanism. The second mechanism involved an abrupt increase in upstream velocity and shear rate, followed by nitro-arginine sensitive dilation. To test whether the abrupt increase in shear could be caused by other agents known to cause edema, remote responses to histamine and thrombin were tested. Each caused an abrupt increase in velocity followed by nitro-arginine-sensitive dilation. This study shows that VEGF or agents that increase permeability can initiate an upstream velocity increase with dilation that recruits flow to the network; this is in addition to simultaneous gap junction-mediated dilation.

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“…These differences probably exist due to donor heterogeneity in HUVEC collection as well as differences in culture conditions. Indeed, Moy et al (2006) described marked differences in functional properties and second messenger activity in HUVEC that were maintained in culture media of varying formulations. Specifically, it was observed that substantial differences existed in the effect of elevated cAMP levels on cell permeability in HUVEC maintained in media recommended by Cambrex Bio Science (as used in the present study) or media less enriched in supplements.…”

Section: Epac1 In a 2b Ar-mediated Erk1/2 Activation In Ec 1197mentioning

confidence: 99%

Cyclic AMP-Dependent, Protein Kinase A-Independent Activation of Extracellular Signal-Regulated Kinase 1/2 Following Adenosine Receptor Stimulation in Human Umbilical Vein Endothelial Cells: Role of Exchange Protein Activated by cAMP 1 (Epac1)

Fang¹,

Olah²

2007

J Pharmacol Exp Ther

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A critical process in angiogenesis is endothelial cell proliferation, which requires activation of extracellular signal-regulated kinase (ERK)1/2. This study analyzed the pathway responsible for adenosine-induced ERK1/2 phosphorylation in human umbilical vein endothelial cells (HUVEC). Characterization with adenosine receptor (AR) agonists and antagonists and the AR mRNA profile demonstrated that stimulation of the A 2B AR can mediate ERK1/2 phosphorylation in HUVEC. The lack of sensitivity of A 2B AR-mediated ERK1/2 phosphorylation to 3- -8220) indicated that protein kinase C stimulation is not required. The response did not involve transactivation of receptors for epidermal growth factor or vascular endothelial growth factor (VEGF). The A 2B AR-mediated response required functional G ␣s and was mimicked by forskolin and 8-bromoadenosine 3Ј,5Ј-cyclic monophosphate. However, ERK1/2 phosphorylation induced by A 2B AR stimulation and forskolin was insensitive to protein kinase A inhibitors. It was hypothesized that the A 2B AR-mediated ERK1/2 activation may involve exchange protein activated by cAMP (Epac), a cAMP-activated guanine nucleotide exchange factor for Rap GTPases. Reverse Transcription-polymerase chain reaction analysis detected Epac1 but not Epac2 in HUVEC. 8-(p-Chlorophenylthio)-2Ј-O-methyladenosine-3Ј,5Ј-cyclic monophosphate (8CPT-2Me-cAMP), an Epac activator, stimulated ERK1/2 phosphorylation. Overexpression of Epac1 enhanced A 2B AR-mediated and forskolin-induced ERK1/2 phosphorylation, whereas response to VEGF was unaffected. Inhibition of Epac1 expression with small interfering RNA substantially reduced A 2B AR-mediated and forskolin-induced ERK1/2 phosphorylation and abolished that by 8CPT-2Me-cAMP. A 2B AR stimulation and forskolin activated Rap1. Expression of a dominant-negative Ras protein did not affect either forskolin-induced or A 2B AR-mediated ERK1/2 phosphorylation. In summary, Epac1 activation in HUVEC results in ERK1/2 activation, and this protein, at least in part, mediates response to the physiologically relevant event of A 2B AR stimulation.Intracellular accumulation of the second messenger molecule cAMP may have positive or negative effects on cell proliferation. These qualitatively different responses are frequently consistent with the effects of cAMP on ERK1/2, which is a key regulator of cell proliferation and may reflect cell type-dependent

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Growth factor- and heparin-dependent regulation of constitutive and agonist-mediated human endothelial barrier function

Cited by 5 publications

References 31 publications

VEGF-C Alters Barrier Function of Cultured Lymphatic Endothelial Cells Through a VEGFR-3-Dependent Mechanism

VEGF-C Alters Barrier Function of Cultured Lymphatic Endothelial Cells Through a VEGFR-3-Dependent Mechanism

Downstream Exposure to Growth Factors Causes Elevated Velocity and Dilation in Arteriolar Networks

Cyclic AMP-Dependent, Protein Kinase A-Independent Activation of Extracellular Signal-Regulated Kinase 1/2 Following Adenosine Receptor Stimulation in Human Umbilical Vein Endothelial Cells: Role of Exchange Protein Activated by cAMP 1 (Epac1)

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