Dataset Information

Endothelial [Ca2+]i and caveolin-1 antagonistically regulate eNOS activity and microvessel permeability in rat venules.

ABSTRACT:

Aims

In this study, we investigated the mechanisms by which caveolin-1 (CAV) inhibits increases in permeability induced by platelet activating factor (PAF) and elucidated the relationship between the endothelial intracellular Ca(2+) concentration ([Ca(2+)](i)) and CAV in regulating endothelial nitric oxide synthase (eNOS) activity and microvessel permeability in intact microvessels.

Methods and results

Experiments were conducted in individually perfused mesenteric venules in Sprague-Dawley rats. Permeability was determined by measuring hydraulic conductivity (Lp). Endothelial [Ca(2+)](i) and nitric oxide (NO) production were measured in fura-2- and DAF-2-loaded microvessels. Perfusion of the CAV scaffolding domain, AP-CAV, at 1 microM for 30 min did not affect PAF-induced increases in endothelial [Ca(2+)](i) but significantly attenuated PAF-induced NO production from 143 +/- 2 to 110 +/- 3% of control fluorescence intensity (FI). The PAF-induced Lp increase was correlatively reduced from a mean peak value of 7.5 +/- 0.9 to 1.9 +/- 0.5 times that of the control. Increasing extracellular [Ca(2+)] that potentiated PAF-induced peak [Ca(2+)](i) from 500 to 1225 nM augmented NO production to 193 +/- 13% and further increased Lp to 17.3 +/- 1.6 times the control value. More importantly, enhanced Ca(2+) influx restored the reduced NO production and Lp by AP-CAV with NO FI at 149% and Lp at 7.7 +/- 1.1 times the control value.

Conclusion

Our results indicate that eNOS inhibition and reduced NO production contribute to the inhibitory action of AP-CAV on PAF-induced increases in permeability. CAV and endothelial [Ca(2+)](i) antagonistically regulate eNOS activity in intact microvessels, and the level of produced NO is the key determinant of the degree of permeability increases during inflammation.

SUBMITTER: Zhou X

PROVIDER: S-EPMC2895537 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature

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Publications

Endothelial [Ca2+]i and caveolin-1 antagonistically regulate eNOS activity and microvessel permeability in rat venules.

Zhou Xueping X He Pingnian P

Cardiovascular research 20100115 2

<h4>Aims</h4>In this study, we investigated the mechanisms by which caveolin-1 (CAV) inhibits increases in permeability induced by platelet activating factor (PAF) and elucidated the relationship between the endothelial intracellular Ca(2+) concentration ([Ca(2+)](i)) and CAV in regulating endothelial nitric oxide synthase (eNOS) activity and microvessel permeability in intact microvessels.<h4>Methods and results</h4>Experiments were conducted in individually perfused mesenteric venules in Sprag ...[more]

PMID: 20080986

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Project description:Vascular remodeling has been implicated in many inflammation-involved diseases. This study aims to investigate the microvascular remodeling-associated alterations in cell-cell adhesion and cytoskeleton reactions to inflammatory stimuli and their impact on microvessel permeability. Experiments were conducted in individually perfused rat mesenteric venules. Microvessel permeability was determined by measuring hydraulic conductivity (Lp), and endothelial intracellular calcium concentration, [Ca(2+)](i), was measured in fura-2-perfused vessels. Alterations in VE-cadherin and F-actin arrangement were examined by confocal imaging. Vascular wall cellular composition and structural changes were evaluated by electron microscopy. Vessels exposed to platelet activating factor (PAF) on day 1 were reevaluated 3 days later in rats that had undergone survival surgery. Initial PAF exposure and surgical disturbance increased microvascular wall thickness along with perivascular cell proliferation and altered F-actin arrangement. Although basal permeability was not changed, upon reexposure to PAF, peak endothelial [Ca(2+)](i) was augmented and the peak Lp was 9.3 ± 1.7 times higher than that of day 1. In contrast to patterns of PAF-induced stress fiber formation and VE-cadherin redistribution observed in day 1 vessels, the day 4 vessels at the potentiated Lp peak exhibited wide separations of VE-cadherin between endothelial cells and striking stress fibers throughout the vascular walls. Confocal images and ultrastructural micrographs also revealed that the largely separated VE-cadherin and endothelial gaps were completely covered by F-actin bundles in extended pericyte processes at the PAF-induced Lp peak. These results indicate that inflammation-induced vascular remodeling increased endothelial susceptibility to inflammatory stimuli with augmented Ca(2+) response resulting in upregulated contractility and potentiated permeability increase. Weakened adhesions between the endothelial cells and contractile mechanisms are both involved in increasing permeability in the intact microvessels and are aggravated during remodeling. The perivascular cells play important roles in stabilizing the microvessel wall, while lessening an otherwise much greater magnitude of leakage during cytoskeletal contraction.

Dataset Information

Endothelial [Ca2+]i and caveolin-1 antagonistically regulate eNOS activity and microvessel permeability in rat venules.

Aims

Methods and results

Conclusion

Publications

Endothelial [Ca2+]i and caveolin-1 antagonistically regulate eNOS activity and microvessel permeability in rat venules.

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