The endothelium-dependent vasodilator effect of acetylcholine: Characterization of the endothelial relaxing factor with inhibitors of arachidonic acid metabolism

Förstermann, Ulrich; Neufang, B.

doi:10.1016/0014-2999(84)90190-0

Cited by 69 publications

(31 citation statements)

References 21 publications

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“…Our results indicate that these cells lose their responsiveness to these agonists after a few days in culture, confirming a previous finding (Cocks et al, 1985). However, these agonists do release prostacyclin and/or EDRF from intact vascular tissue in vitro and in vivo (Furchgott & Zawadski, 1980;Beetens et al, 1982;Gryglewski, 1982;Cocks & Angus, 1983;Forstermann & Neufang, 1984). Paf-acether did not release prostacyclin or EDRF from the column.…”

Section: Discussionsupporting

confidence: 89%

“…Unlike cyclo-oxygenase, simple additions of substrate are not enough for the lipoxygenase to generate its products (Borgeat & Samuelsson, 1979). Therefore the small and erratic release of EDRF following stimulation with high concentrations of AA neither confirmed nor excluded the possibility that EDRF might be a metabolite of this fatty acid (Furchgott & Zawadzki, 1980;Forstermann & Neufang, 1984;Peach et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

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Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Gryglewski

Moncada

Palmer

1986

British J Pharmacology

217

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A cascade superfusion technique has been developed for the differential bioassay of prostacyclin and endothelium-derived relaxing factor (EDRF) released from porcine aortic endothelial cells cultured on microcarriers, packed into a column and perfused. 2 Bradykinin (Bk; 20-100 nM) released prostacyclin (9.6 ± 1.5 nm per 106 cells; mean ± s.e.mean, n = 9) and prostaglandin E2 (PGE2; 2.1 ± 0.6 nm per 10' cells) from the column measured by relaxation of strips of bovine coronary artery (BCA) and rabbit mesenteric or coeliac artery, respectively. The presence of these prostanoids in the effluent was confirmed by specific radioimmunoassays. 3 A23187 (500-2000nM) also released both prostacyclin and PGE2 from the cells. This release was long-lasting and not reproducible. 4 Bk (20-100 nM) and A23187 (30-300 nM) released EDRF from the column. This was detected in a cascade of four rabbit aortic strips (RbA), denuded of endothelium and contracted with U46619 or phenylephrine. The relaxation of the RbA strips caused by EDRF was progressively attenuated down the cascade (half-life < 7 s) and was not affected by indomethacin. 5 EDRF and prostacyclin could be differentially bioassayed in a cascade of alternating RbAs and BCAs as prostacyclin did not relax RbAs and the time delay to the BCAs destroys EDRF. EDRF could be bioassayed on its own when the endothelial cells were treated with indomethacin. 6 5-Hydroxytryptamine 0.2, noradrenaline 1.0, platelet-activating factor (Paf-acether) 1.0, formylmethionyl-leucyl-phenylalanine 1.0, acetylcholine 0.5, bethanecol 0.5, adenosine diphosphate 0.25 and angiotensin II 0.1 gM did not release either prostanoids or EDRF from the column.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Gryglewski

Moncada

Palmer

1986

British J Pharmacology

217

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“…This observation suggests that PGI2 released by Bk and A23187 does not contribute to the peak of the vasodilator response. Similarly, EDRF responses are known to occur independently of the action of PGI2 (Furchgott & Zawadzski, 1980;Forstermann & Neufang, 1984a).…”

Section: Discussionmentioning

confidence: 99%

Vasodilator actions of acetylcholine, A23187 and bradykinin in the guinea‐pig isolated perfused heart are independent of prostacyclin

Stewart

Piper

1988

British J Pharmacology

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1 The involvement of prostacyclin (PGI2) in the vasodilator responses to acetylcholine (ACh), A23187 and bradykinin (Bk) has been investigated in guinea-pig, isolated, Krebs-perfused hearts. 2 ACh (0.01-lOnmol), A23187 (0.1-1.Onmol) and Bk (0.3-lOpmol) each elicited dose-related and shortlasting (-2 min) reductions in perfusion pressure. Larger maximal responses were obtained in preparations with coronary vascular tone elevated by platelet-activating factor (100 pmol) than in preparations at basal perfusion pressure. 3 Bk and A23187 elicited dose-related increases in the generation of PGI2 as measured by its chemically-stable breakdown product, 6-oxo-PGFg. Indomethacin (2.8 uM) prevented both basal and the stimulated generation of 6-oxo-PGF,., whereas the magnitudes of the vasodilator responses were unaffected. 4 Attempts to identify the release of vasodilator materials by on-line superfusion bioassay of cardiac effluent were unsuccessful, indicating a possible role for a labile vasodilator such as endothelium-dependent relaxing factor (EDRF). In addition, the inhibitors of EDRF action/ production, mepacrine (3yM) or diethylcarbamazine (300pM), attenuated vasodilator responses to ACh without altering those to the endothelium-independent vasodilator, verapamil (1 nmol). 5 Haemoglobin (10pM) reduced vasodilator responses to ACh, Bk and verapamil and abolished those induced by A23187. Inhibition of the endothelium-independent vasodilator, verapamil, was significantly less than that for the other compounds. 6 The present data indicate the existence of an indomethacin-resistant vasodilator mechanism in the coronary microcirculation in response to ACh, A23187 and Bk. EDRF is a candidate for mediating these responses; however, a direct vasodilator action of these substances cannot be excluded.

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“…In the vasculature, insulin and acetylcholine (ACh) regulate the release of NO (Förstermann and Neufang, 1984;Zeng and Quon, 1996) and the regulation of mRNA expression of constitutive eNOS (Papapetropoulos et al, 1999;Kuboki et al, 2000). To verify that the effects on osteogenesis were truly based on generation of NO, we conducted another experimental set, in which we treated ESCs with these two upstream NO regulators during days 3 to 5.…”

Section: Upstream No Effectorsmentioning

confidence: 99%

NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic stem cell osteogenic differentiation

Ding

Keller

Martinez

et al. 2012

Journal of Cell Science

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SummaryNitric oxide (NO) has been shown to play a crucial role in bone formation in vivo. We sought to determine the temporal effect of NO on murine embryonic stem cells (ESCs) under culture conditions that promote osteogenesis. Expression profiles of NO pathway members and osteoblast-specific markers were analyzed using appropriate assays. We found that NO was supportive of osteogenesis specifically during an early phase of in vitro development (days 3-5). Furthermore, ESCs stably overexpressing the inducible NO synthase showed accelerated and enhanced osteogenesis in vitro and in bone explant cultures. To determine the role of NO in early lineage commitment, a stage in ESC differentiation equivalent to primitive streak formation in vivo, ESCs were transfected with a T-brachyury-GFP reporter. Expression levels of T-brachyury and one of its upstream regulators, b-catenin, the major effector in the canonical Wnt pathway, were responsive to NO levels in differentiating primitive streak-like cells. Our results indicate that NO may be involved in early differentiation through regulation of b-catenin and T-brachyury, controlling the specification of primitive-streak-like cells, which may continue through differentiation to later become osteoblasts.

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The endothelium-dependent vasodilator effect of acetylcholine: Characterization of the endothelial relaxing factor with inhibitors of arachidonic acid metabolism

Cited by 69 publications

References 21 publications

Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Vasodilator actions of acetylcholine, A23187 and bradykinin in the guinea‐pig isolated perfused heart are independent of prostacyclin

NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic stem cell osteogenic differentiation

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