Neovascular responses induced by cultured aortic endothelial cells

Harris‐Hooker, Sandra; Gajdusek, Corinne M.; Wight, Thomas N.; Schwartz, Stephen M.

doi:10.1002/jcp.1041140308

Cited by 66 publications

(23 citation statements)

References 31 publications

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“…L-NAME (nmol/disc) Figure 4 Effect of increasing amounts of (a) N0-monomethyl-L- Hooker et al (1983) showed that the vascular density under the disc containing 38.7 nmol of dexamethasone increased by 31.5 ± 4.4%, n = 6, P<0.01. At 116.1 nmol/disc, the angiogenic effect of dexamethasone was reduced to 18.2 ± 4.6%, n = 7 and at 348.4 and 1161 nmol/disc it caused a 24.8 ± 9.5%, n = 6 (P<0.05) and 65.0 ± 4.9%, n = 7 (P< 0.01) decrease in collagenous protein biosynthesis ( Figure 6a).…”

Section: Resultsmentioning

confidence: 99%

Evidence that nitric oxide is an endogenous antiangiogenic mediator

Pipili-Synetos¹,

Sakkoula

Haralabopoulos

et al. 1994

British J Pharmacology

155

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The involvement of nitric oxide (NO) in the regulation of angiogenesis was examined in the in vivo system of the chorioallantoic membrane (CAM) of the chick embryo and in the matrigel tube formation assay. Sodium nitroprusside (SNP) (0.37–28 nmol/disc), which releases NO spontaneously, caused a dose‐dependent inhibition of angiogenesis in the CAM in vivo and reversed completely the angiogenic effects of α‐thrombin (6.7 nmol/disc) and the protein kinase C (PKC) activator 4‐β‐phorbol‐12‐myristate‐13‐acetate (PMA) (0.97 nmol/disc). In addition, SNP (28 × 10−6 m) stimulated the release of guanosine 3′‐5′‐cyclic monophosphate (cyclic GMP) from the CAM in vitro. In the matrigel tube formation assay, an in vitro assay of angiogenesis, both SNP (1–3 × 10−6 m) and the cell permeable cyclic GMP analogue, Br‐cGMP (0.3–1.0 × 10−3 m) reduced tube formation. The inhibitors of NO synthase, NG‐monomethyl‐l‐arginine (l‐NMMA) (3.8–102 nmol/disc) and NG‐nitro‐l‐arginine methylester (l‐NAME) (1.3–34.2 nmol/disc) stimulated angiogenesis in the CAM in vivo, in a dose‐dependent fashion. d‐NMMA and d‐NAME on the other hand had no effect on angiogenesis in this system. l‐Arginine (10.9 nmol/disc), although it had a modest antiangiogenic effect by itself, was capable of abolishing the angiogenic effects of l‐NMMA (34.2 nmol/disc) and of l‐NAME (3.8 nmol/disc). Dexamethasone, an inhibitor of the induction of NO synthase, at 0.2–116.1 nmol/disc, stimulated angiogenesis in the CAM, whereas at 348.4–1161 nmol/disc it inhibited this process. Combination of 38.7 nmol/disc dexamethasone with l‐NAME (9.3 nmol/disc) resulted in a potentiation of the angiogenic effect of the former. It appears therefore that both the constitutive and the inducible NO synthase may contribute to the NO‐mediated inhibition of angiogenesis. Superoxide dismutase (SOD), which prevents the destruction of NO, at 300 i.u./disc had a modest antiangiogenic effect in the CAM, by itself. In addition, SOD, prevented α‐thrombin (6.7 nmol/disc) and PMA (0.97 nmol/disc) from stimulating angiogenesis in the CAM. These results suggest that NO may be an endogenous antiangiogenic molecule of pathophysiological importance.

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Section: Resultsmentioning

confidence: 99%

Evidence that nitric oxide is an endogenous antiangiogenic mediator

Pipili-Synetos¹,

Sakkoula

Haralabopoulos

et al. 1994

British J Pharmacology

155

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“…The windows were covered and the eggs were incubated until day 11, before assessment of angiogenesis. Therefore, the eggs were flooded with 10% buffered formalin, the discs were removed, and the eggs were kept at room temperature for at least 2 h. A large area around the discs was cut out and placed on a glass slide, and the vascular density index was measured by the method of Harris-Hooker et al [33]. Briefly, a grid containing three concentric circles of 6-, 8-and 10-mm diameter was positioned on the surface of the CAM previously covered by the disc.…”

Section: Cam Assaymentioning

confidence: 99%

Soluble N-cadherin fragment promotes angiogenesis

Derycke

Morbidelli

Ziche

et al. 2006

Clin Exp Metastasis

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Endothelial cells express two dependent intercellular adhesion molecules: vascular endothelial (VE)-cadherin, specific for endothelial cells, and Ncadherin, also present in neuronal, lens, skeletal and heart muscle cells, osteoblasts, pericytes and fibroblasts. While there exists a vast amount of evidence that VE-cadherin promotes angiogenesis, the role of N-cadherin still remains to be elucidated. We found that a soluble 90-kDa fragment N-cadherin promotes angiogenesis in the rabbit cornea assay and in the chorioallantoic assay when cleaved enzymatically from the extracellular domain of N-cadherin. Soluble N-cadherin stimulates migration of endothelial cells in the wound healing assay and stimulates phosphorylation of extracellular regulated kinase. In vitro experiments with PD173074 and knock-down of N-cadherin and fibroblast growth factor (FGF)-receptor, showed that the pro-angiogenic effect of soluble N-cadherin is N-cadherin-and FGF-receptor-dependent. Our results suggest that soluble N-cadherin stimulates migration of endothelial cells through the FGF-receptor.

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“…The application of stereological principles to the analysis of microcirculation represents a useful tool for obtaining quantitative information [22]. Several stereological techniques have been described to quantitate an angiogenic response in the CAM [23][24][25][26]. A VDI was calculated by counting the number of intercepts of vessels with concentric circles of a grid, which was placed over the formalin-fixed CAM, or over a photograph of the excised membranes.…”

Section: Discussionmentioning

confidence: 99%

The Chick Embryo Chorioallantoic Membrane as a Model to Investigate the Angiogenic Properties of Human Endometrium

Maas¹,

Noble²,

Dunselman³

et al. 1999

Gynecol Obstet Invest

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The chick embryo chorioallantoic membrane (CAM) is an established in vivo angiogenesis assay. The aim of our study was to assess the angiogenic properties of endometrium and to quantitate the vascular response in an accurate way. Samples of proliferative endometrium (n = 17) and control mouse skin tissue (n = 8) were explanted onto the CAM at day 10 of incubation. Additional controls consisted of normal unmanipulated CAM (n = 12). Four days after grafting, photographs of the explant and the surrounding area were taken in ovo to measure the vascular density index (VDI). The VDI is a stereological estimate of vessel number and length, which was obtained by counting the intersections of vessels with a circular grid superimposed on a computerized image. Endometrium caused a significant increase in VDI as compared to both unmanipulated CAM (p < 0.001) and skin tissue as a control (p < 0.007). The intra-observer variability was 5.2%. This study demonstrates that the CAM assay is a suitable model to assess the angiogenic properties of endometrium. Furthermore, it allows detailed quantitation of the vascular response in an objective and reproducible way. Our findings suggest the CAM to be a promising model to study the role of angiogenesis in both normal human endometrium and diseases involving the endometrium.

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Neovascular responses induced by cultured aortic endothelial cells

Cited by 66 publications

References 31 publications

Evidence that nitric oxide is an endogenous antiangiogenic mediator

Evidence that nitric oxide is an endogenous antiangiogenic mediator

Soluble N-cadherin fragment promotes angiogenesis

The Chick Embryo Chorioallantoic Membrane as a Model to Investigate the Angiogenic Properties of Human Endometrium

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