<i>In vitro</i> studies of release of adenine nucleotides and adenosine from rat vascular endothelium in response to α<sup>1‐</sup>adrenoceptor stimulation

Shinozuka, Kazumasa; Hashimoto, Michio; Masumura, Sumio; Bjur, Richard A.; Westfall, David P.; Hattori, Keisuke

doi:10.1111/j.1476-5381.1994.tb17125.x

Cited by 66 publications

(48 citation statements)

References 24 publications

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“…Thus the identification of alternative sources of extracellular nucleotides and the elucidation of mechanisms underlying this nonsynaptic nucleotide release have become significant areas of current investigation. In the studies described in the current article in focus Previous studies determined that multiple types of vascular EC, including human umbilical vein EC (2,12), EC from rat aorta and caudal artery (18), porcine aortic EC (14), and guinea pig cardiac EC (20), can release nucleotides via noncytolytic but otherwise undefined mechanisms. Two types of stimuli were shown to trigger ATP release from these various EC types: mechanical stress during changes in flow and activation of Ca 2ϩ -mobilizing receptors for different vasoactive hormones/neurotransmitters.…”

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confidence: 99%

“…This particular methodological approach was useful in two ways. Previous studies of ATP release from EC utilized "off-line" methods that involved removal of bathing medium at various times following stimulation of the EC with agonists or mechanical stress, followed by enzymatic or HPLC-based analyses of nucleotide content within these extracellular samples (2,12,14,18,20). Although sensitive, such off-line assays suffer from a limited capacity for temporal resolution of transient ATP release events from EC.…”

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confidence: 99%

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Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Dubyak

2002

American Journal of Physiology-Cell Physiology

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confidence: 99%

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confidence: 99%

Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Dubyak

2002

American Journal of Physiology-Cell Physiology

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“…Recently, this idea has been directly demonstrated by the results of studies with cultured cardiac (7) and vascular (5) endothelial cells. Furthermore, most of ATP released from the endothelial cells seems to be rapidly metabolized to adenosine by ectonucleotidase (5).…”

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confidence: 99%

“…The values were expressed as a percentage of the maximum contraction induced in each artery by the 6-min treatment with 1.0 pM noradrenaline. The details of the experiments have been described previously (5). All drugs and reagents were obtained from commercial sources.…”

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confidence: 99%

Participation of Endogenous Noradrenaline in the Release of ATP by High Potassium in the Rat Caudal Artery

Shinozuka

Kunitomo

Shimoura

et al. 1995

Japanese Journal of Pharmacology

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ABSTRACT-Potassium (40 mM) evoked the release of adenine nucleotides and adenosine in rat isolated caudal arteries. This release was abolished by bunazosin. Endogenous noradrenaline was also released. Furthermore, the high potassium-induced purine release was abolished by rubbing the lumen of the vessel. These findings suggest that the high potassium releases endogenous noradrenaline, which subsequently elicits the release of ATP and related purines by acting on a,-adrenoceptors on the endothelial cell. Keywords:ATP release, Noradrenaline, EndotheliumAdenine nucleotides and adenosine are generally considered to be released upon nerve stimulation of a variety of sympathetically innervated tissues. In the caudal arteries of rats, electrical nerve stimulation and an a,-adrenoceptor agonist, methoxamine, have been shown to evoke the release of a large amount of adenine nucleotides and adenosine from extraneuronal sites (1, 2). Sedaa et al. (3) suggested that in rabbit aorta, the source for the adenine nucleotides and adenosine released by methoxamine is primarily the endothelial cells. However, the details of the ATP release from the endothelium have not been sufficiently identified. In the present study, we examined the effect of high potassium on the release of ATP from the rat caudal artery to characterize the release of ATP.The caudal artery (generally about 9 cm) was removed from male Sprague Dawley rats (SLC, Hamamatsu; ranging from 15 to 20 weeks of age) anesthetized with sodium pentobarbital (50 mg/kg, i.p.). In the release experiments, the caudal artery was cut open lengthwise; and for the measurement of contractile activity, the artery was cut into small rings 2 -3 mm in length. Some experiments were carried out with arterial preparations in which the endothelial cell lining was disrupted. This was accomplished by gently rubbing the lumen of the artery with a stainless steel wire. In the release experiment, the whole caudal arterial preparation was suspended in a water-jacketed organ chamber containing 2.0 ml of a modified Krebs solution and allowed to equilibrate for 60 min. The medium was replaced every 3 min in the latter half of the equilibration period. After the equilibration, the bathing solution was rapidly collected (0-min sample: basal overflow for 3 min) by draining the organ chamber. Then the tissue was stimulated for 6 min with 40 mM potassium or 1.0 tM norepinephrine, and the bathing solution was collected twice at 3-min intervals; i.e., 3-min and 6-min samples, respectively. The purines, such as ATP, ADP, AMP and adenosine, and noradrenaline in the sample solution were analyzed by high performance liquid chromatography with fluorescence detection (4) and electrochemical detection (5), respectively. To compare the contractions induced by potassium and noradrenaline, the ring preparations of the caudal artery from four rats were stimulated first with noradrenaline and then with potassium at 30-min intervals. Another four preparations were stimulated with potassium and then with noradrenaline. ...

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“…1,4) Vanadium is a widely distributed element that assumes several experimental oxidation states including that of vanadate (VO 4 Ϫ ). This phosphate analogue is an insulinomimetic, and has been reported to exert beneficial metabolic and cardiovascular effects in several models of diabetes.…”

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confidence: 99%

Effect of Vanadate on ATP-Induced Increase in Intracellular Calcium Ion Levels in Human Umbilical Vein Endothelial Cells

Nejime

Tada

Kagota

et al. 2010

Biological & Pharmaceutical Bulletin

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The vascular endothelium participates in various physiological and pathophysiological responses, including blood flow, angiogenesis, blood coagulation, and permeability to fluids and solutes. Many mediators initiate these diverse functions of endothelial cells. ATP is one endogenous modulator that is involved in nitric oxide production, proliferation, migration, apoptosis, and regulation of permeability in these cells. 1,2) We previously found that ATP increases intracellular calcium ion level ([Ca 2ϩ ] i ) of cultured endothelial cells from the rat caudal artery and regulates intercellular permeability.3) Endothelial cells also release ATP and its metabolites during shear stress, a 1 -adrenoceptor activation, and hypoxia. 1,4) Vanadium is a widely distributed element that assumes several experimental oxidation states including that of vanadate (VO 4 Ϫ ). This phosphate analogue is an insulinomimetic, and has been reported to exert beneficial metabolic and cardiovascular effects in several models of diabetes.5-7) On the other hand, Misurski et al. 8) showed that sodium orthovanadate elicited vasorelaxation of the rat mesenteric vascular bed that was mediated by endothelial-derived nitric oxide and endothelial-derived hyperpolarizing factor (EDHF) but not by vasodilator eicosanoids. Vanadium has recently been added to mineral water as a health supplement.Based on the above evidence, we examined the effect of vanadate on ATP-induced increase in [Ca 2ϩ ] i of human umbilical vein endothelial cells (HUVEC). MATERIALS AND METHODSEndothelial Cell Culture HUVEC purchased from BioWhittaker (Walkersville, MD, U.S.A.) were cultured in EBM-2 medium supplemented with 5% fetal bovine serum, hydrocortisone, endothelial growth factor, epidermal growth factor, human fibroblast growth factor, insulin-like growth factor, and ascorbic acid. Cells were seeded in 57-cm 2 tissue culture dishes (Cell tight C-1; Sumitomo Bakelite Co., Ltd., Osaka, Japan) and incubated at 37°C in a 95% air/5% CO 2 atmosphere. Cells were removed from the dishes using 0.025% trypsin/0. N,NЈ, to PSS to remove CaCl 2 . Primary cultured endothelial cells on type I collagen-coated glass coverslips were incubated with 5 mmol/l of Calcium Green 1/AM at 37°C for 30 min. Cells were rinsed three times and incubated in PSS at 37°C for an additional 15 min in a 95% air/5% CO 2 atmosphere to complete hydrolysis of any intact ester linkages in the intracellular Calcium Green 1/AM. Loaded coverslips were placed onto the stage of an inverted microscope coupled to a Nipkow disk confocal scanner (CSU10; Yokogawa Electric Corp., Tokyo, Japan) and ARGUS-50 imaging system (Hamamatsu Photonics). An argon-krypton laser (Omnichrome, Chino, CA, U.S.A.) emitted an excitation wavelength of 488 nm. Emitted light was collected with a 510-nm long-pass dichroic reflector and 515-nm long-pass emission filter through a planfluor objective (ϫ100 or ϫ20; NAϭ1.4 and 0.5, respectively). Fluorescence images were acquired by ICCD camera (C2400-87; Hamamatsu Photonics) at a resolu...

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In vitro studies of release of adenine nucleotides and adenosine from rat vascular endothelium in response to α^1‐adrenoceptor stimulation

Cited by 66 publications

References 24 publications

Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Participation of Endogenous Noradrenaline in the Release of ATP by High Potassium in the Rat Caudal Artery

Effect of Vanadate on ATP-Induced Increase in Intracellular Calcium Ion Levels in Human Umbilical Vein Endothelial Cells

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In vitro studies of release of adenine nucleotides and adenosine from rat vascular endothelium in response to α1‐adrenoceptor stimulation

Cited by 66 publications

References 24 publications

Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Focus on “Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells”

Participation of Endogenous Noradrenaline in the Release of ATP by High Potassium in the Rat Caudal Artery

Effect of Vanadate on ATP-Induced Increase in Intracellular Calcium Ion Levels in Human Umbilical Vein Endothelial Cells

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In vitro studies of release of adenine nucleotides and adenosine from rat vascular endothelium in response to α^1‐adrenoceptor stimulation