High-Performance Liquid Chromatographic Technique for Detection of a Fluorescent Analogue of ADP-Ribose in Isolated Blood Vessel Preparations

Bobáľová, Janette; Bobáľ, Pavel; Mutafova‐Yambolieva, Violeta N.

doi:10.1006/abio.2002.5667

Cited by 46 publications

(91 citation statements)

References 31 publications

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“…eUp4A was produced by first acidifying 0.2 mM Up4A to pH 4.0 with citrate phosphate buffer, then adding 1 M chloroacetaldehyde, and etheno-derivatizing at 80°C for 40 min (23). eUp4A was used as a standard to analyze Up4A in tissue superfusates.…”

Section: Methodsmentioning

confidence: 99%

Uridine adenosine tetraphosphate is a novel neurogenic P2Y1 receptor activator in the gut

Durnin

Hwang

Kurahashi

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Section: Methodsmentioning

confidence: 99%

Uridine adenosine tetraphosphate is a novel neurogenic P2Y1 receptor activator in the gut

Durnin

Hwang

Kurahashi

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Time controls parallel to drug treatment protocols were carried out with each group of experiments. Samples were analyzed for nucleotide/ nucleoside contents by HPLC techniques with fluorescence detection as described previously (5)(6)(7)31).…”

Section: Overflow Experimentsmentioning

confidence: 99%

“…Briefly, 100 l of a citrate phosphate buffer (pH 4.0) were added to 200 l of the superfusate sample in a 1.5 ml Eppendorf tube. Chloroacetaldehyde was synthesized according to a method modified from Secrist et al (30) and Levitt et al (22) and described previously (5). Ten microliters of 2-chloroacetaldehyde were added to the samples in a fume hood; the samples were heated for 40 min at 80°C in a dry bath incubator (Fisher Scientific) to produce 1,N 6 -etheno-nucleotides and 1,N 6 -etheno-nucleosides.…”

Section: Sample Preparation and Hplc Assay Of Etheno-nucleotides And mentioning

confidence: 99%

“…Ten microliters of 2-chloroacetaldehyde were added to the samples in a fume hood; the samples were heated for 40 min at 80°C in a dry bath incubator (Fisher Scientific) to produce 1,N 6 -etheno-nucleotides and 1,N 6 -etheno-nucleosides. The liquid chromatographic system used throughout this study was an HP1100 LC module system (Agilent Technologies, Wilmington, DE) as described previously (5,31). The mobile phase comprised of 0.1 mol/l KH 2PO4 (pH 6.0) as eluent A; eluent B consisted of 35% methanol and 65% eluent A. Gradient elution was employed according to the following linear program: time 0, 0% eluent B; 18 min, 100% eluent B.…”

Section: Sample Preparation and Hplc Assay Of Etheno-nucleotides And mentioning

confidence: 99%

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β-NAD is a novel nucleotide released on stimulation of nerve terminals in human urinary bladder detrusor muscle

Breen

Smyth²,

Yamboliev³

et al. 2006

American Journal of Physiology-Renal Physiology

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“…Derivatized samples were processed through a gradient HP1100 liquid chromatography module system equipped with a fluorescence detector as described (44). Each 1,N 6 -ethenopurine was quantified against known standards.…”

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confidence: 99%

β-Nicotinamide adenine dinucleotide is an inhibitory neurotransmitter in visceral smooth muscle

Mutafova‐Yambolieva

Hwang

Hao

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Peripheral inhibitory nerves are physiological regulators of the contractile behavior of visceral smooth muscles. One of the transmitters responsible for inhibitory neurotransmission has been reputed to be a purine, possibly ATP. However, the exact identity of this substance has never been verified. Here we show that ␤-nicotinamide adenine dinucleotide (␤-NAD), an inhibitory neurotransmitter candidate, is released by stimulation of enteric nerves in gastrointestinal muscles, and the pharmacological profile of ␤-NAD mimics the endogenous neurotransmitter better than ATP. Levels of ␤-NAD in superfusates of muscles after nerve stimulation exceed ATP by at least 30-fold; unlike ATP, the release of ␤-NAD depends on the frequency of nerve stimulation. ␤-NAD is released from enteric neurons, and release was blocked by tetrodotoxin or -conotoxin GVIA. ␤-NAD is an agonist for P2Y1 receptors, as demonstrated by receptor-mediated responses in HEK293 cells expressing P2Y1 receptors. Exogenous ␤-NAD mimics the effects of the enteric inhibitory neurotransmitter. Responses to ␤-NAD and inhibitory junction potentials are blocked by the P2Y1-selective antagonist, MRS2179, and the nonselective P2 receptor antagonists, pyridoxal phosphate 6-azophenyl-2,4-disulfonic acid and suramin. Responses to ATP are not blocked by these P2Y receptor inhibitors. The expression of CD38 in gastrointestinal muscles, and specifically in interstitial cells of Cajal, provides a means of transmitter disposal after stimulation. ␤-NAD meets the traditional criteria for a neurotransmitter that contributes to enteric inhibitory regulation of visceral smooth muscles.enteric nervous system ͉ gastrointestinal motility ͉ P2Y receptor ͉ purinergic neurotransmission ͉ interstitial cells of Cajal

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High-Performance Liquid Chromatographic Technique for Detection of a Fluorescent Analogue of ADP-Ribose in Isolated Blood Vessel Preparations

Cited by 46 publications

References 31 publications

Uridine adenosine tetraphosphate is a novel neurogenic P2Y1 receptor activator in the gut

Uridine adenosine tetraphosphate is a novel neurogenic P2Y1 receptor activator in the gut

β-NAD is a novel nucleotide released on stimulation of nerve terminals in human urinary bladder detrusor muscle

β-Nicotinamide adenine dinucleotide is an inhibitory neurotransmitter in visceral smooth muscle

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