An assessment of the antagonistic activity of reactive blue 2 at P1- and P2-purinoceptors: supporting evidence fro purinergic innervation of the rabbit portal vein

Reilly, William M.; Saville, Valerie L.; Burnstock, Geoffrey

doi:10.1016/0014-2999(87)90632-7

Cited by 47 publications

(29 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…RB has been used as a blocker of ATP receptors [9,10], but there are also reports that RB blocks some channels and receptors [11][12][13][14][15]. Besides these inhibitory actions of RB, we here demonstrated that RB had a stimulatory action on the PLC cascade.…”

Section: ؉supporting

confidence: 50%

See 1 more Smart Citation

Reactive Blue 2 Induces Calcium Oscillations in HeLa Cells.

Okuda

Furuya²,

Kiyohara³

2001

JJP

View full text Add to dashboard Cite

Calcium ion is an intracellular second messenger important for cellular functions in a variety of cells. In many cell types, transient and repetitive increases in the intracellularHere we found that the application of RB induced Ca 2ϩ oscillations in HeLa cells. We also found that RB-induced Ca 2ϩ oscillations had unique characteristics different from the other receptor-activated Ca 2ϩ oscillations reported in HeLa cells [6][7][8]. Materials and MethodsHeLa cells were grown in DME/F12 (SIGMA, MO, USA) supplemented with 10% fetal bovine serum, 50 units/ml penicillin, and 50 g/ml streptomycin. Cells were cultured on glass coverslips (25-mm diameter, MATSUNAMI #1) in a humidified atmosphere (5% CO 2 , 95% air) at 37°C.To introduce fura-2 into the cells, they were incubated with 2 M fura-2 AM (acetoxymethyl ester; Dojindo, Kumamoto, Japan) and 0.2% cremophor EL (SIGMA) for 50 min at 37°C. Coverslips bearing the

show abstract

Section: ؉supporting

confidence: 50%

“…Reactive Blue 2 (RB) has been used as an antagonist of ATP receptors (P2-type purinergic receptors) [9,10]. RB, however, is a histochemical dye containing anthraquinone and has been reported to have other effects on cells and enzymes.…”

mentioning

confidence: 99%

Reactive Blue 2 Induces Calcium Oscillations in HeLa Cells.

Okuda

Furuya²,

Kiyohara³

2001

JJP

View full text Add to dashboard Cite

show abstract

“…The most widely studied P2 purinoceptor antagonist, suramin, does not discriminate between the known P2 receptor subtypes (Cusack, 1993). Cibacron blue, an anthraquinone sulphonic acid derivative formerly called reactive blue 2, reportedly shows selectivity for P2y receptors over P2x receptors (Burnstock & Warland, 1987;Reilly et al, 1987), at least within a limited concentration-range. Another sulphonic acid derivative, pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) antagonizes P2x purinoceptor-mediated responses in both the guinea-pig and rabbit vas deferens (Lambrecht et al, 1992;McLaren et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

The use of antagonists to characterize the receptors mediating depolarization of the rat isolated vagus nerve by α,β‐methylene adenosine 5′‐triphosphate

Trezise

Kennedy

Humphrey

1994

British J Pharmacology

View full text Add to dashboard Cite

We have previously found that the Pu-purinoceptor agonist, x,p-methylene adenosine 5'-triphosphate (x,p-methylene ATP), depolarizes the rat cervical vagus nerve, measured with a 'grease-gap' extracellular recording technique. This effect was attenuated by the P2 purinoceptor antagonist, suramin. In the present study we have investigated in more detail the antagonism produced by suramin and have also investigated the actions of two other putative P2 purinoceptor antagonists, cibacron blue and pyridoxalphosphate-6-azophenyl-2', 5'-disulphonic acid (iso-PPADS). Furthermore, we have studied the interactions between suramin and cibacron blue or iso-PPADS in an attempt to determine whether these antagonists act at a common receptor site. 2 Suramin (1 x 10-5-1 X 10-4M) produced reversible, concentration-related rightward displacements of the concentration-effect curve to a,-methylene ATP. Schild analysis of this antagonism yielded a pA2 value of 5.90 with a slope value of 0.47.3 Cibacron blue (3 X 10-5_ x 10-4 M) also antagonized depolarizations induced by op-methylene ATP. The antagonistic effects of cibacron blue were slow to reach equilibrium but could be readily reversed on washout. At low concentations for antagonism, cibacron blue (1 x 10-5 M and 3 x 10-5 M) produced enhancement of the maximal response to a,-methylene ATP. At the highest concentration tested (1 x 10-4 M) the concentration-effect curve to ux,1-methylene ATP was shifted to the right in a parallel manner, yielding a pKB estimate of 4.96.4 Iso-PPADS (1 X 10-6 1 X 1i-M) produced a concentration-related depression in the maxima of the concentration-effect curves to a,-methylene ATP. Analysis of these data by a double reciprocal plot yielded a pKB estimate of 6.02. This profile of insurmountable antagonism could not be attributed to irreversible binding of iso-PPADS to the receptor since the effect of iso-PPADS could be reversed on washing, albeit slowly. 5 In the presence of suramin (1 x 10-4 M), cibacron blue (1 x 10-4 M) produced no further rightward displacement of the a,p-methylene ATP concentration-effect curve. The mean agonist concentrationratios in the presence of suramin or cibacron blue alone (11.7 and 10.3, respectively) were not significantly different from the mean concentration-ratio in the presence of both antagonists (11.8). This finding suggests that high concentrations of a,p-methylene ATP activate a receptor population which is resistant to blockade by either antagonist. 6 The antagonistic effect of iso-PPADS (1 x 10-M) was partially attenuated by suramin (1 x I0-4 M).It is possible that this interaction reflects a slow dissociation of iso-PPADS from the receptor with which suramin and a4,-methylene ATP interact.7 Suramin, cibacron blue or iso-PPADS had no marked effect on depolarization produced by 5-hydroxytryptamine (5-HT, 1 x 10-7-3 x 1-0 M), indicating their specificity in antagonizing responses to a,-methylene ATP.8 Responses to a,-methylene ATP were not antagonized by 8-para-sulphophenyltheophylline (3 x 10-' M), ondansetron (...

show abstract

“…The vasodilator response to ATP in the rabbit mesenteric artery (Mathieson & Burnstock, 1985) and the rabbit portal vein (Reilly et al, 1987) is mediated via P2y-purinoceptors located directly on the smooth muscle not on the endothelium, although NA in these arteries produces a vasoconstrictor response. Ramme and coworkers suggest that in the resistance vessels in the smaller branches of the rabbit mesenteric artery, ATP is the sole neurotransmitter acting postjunctionally upon stimulation of the sympathetic nerves and that the NA released concomitantly acts entirely on prejunctional a2-adrenoceptors to modulate transmitter release (Ramme et al, 1987).…”

Section: Basal Tone Preparationsmentioning

confidence: 99%

“…Thus, exogenous ATP in the rabbit mesenteric artery may be acting upon receptors which would be stimulated physiologically by sensory rather than sympathetic nerves. The rabbit portal vein is innervated by sympathetic nerves but is also supplied with non-sympathetic (purinergic) nerves which release ATP and mediate vasodilatation (Burnstock et al, 1979;Reilly et al, 1987;Su, 1987). These non-adrenergic nerves are the source of approximately 50% of the purines released by electrical field stimulation (Levitt & Westfall, 1982) and may account for the presence of the P2y-purinoceptor on the smooth muscle in this vessel.…”

Section: Basal Tone Preparationsmentioning

confidence: 99%

Vasodilator response of coronary smooth muscle to the sympathetic co‐transmitters noradrenaline and adenosine 5′‐triphosphate

Corr

Burnstock

1991

British J Pharmacology

Self Cite

View full text Add to dashboard Cite

1 Vasodilator and vasoconstrictor responses to noradrenaline (NA), adenosine and adenosine 5'-triphosphate (ATP) were examined in isolated ring segments of the left anterior descending coronary artery of the rabbit in the absence of endothelium.2 NA caused dose-dependent relaxation of potassium-constricted vessels in the absence of aladrenoceptor blockade, with a pD2 of 5.96 + 0.21. This was inhibited by 1 gM propranolol. Constrictor responses of vessels at baseline tension were only seen at concentrations greater than 1 mm, and reached a maximum of 6% of the contraction to 30 mm KCI. 3 ATP caused relaxation of the potassium-constricted ring segments in a dose-dependent manner, although a transient constriction often preceded the relaxation. Adenosine was equipotent with ATP in producing relaxation; this was significantly inhibited by the Pl-purinoceptor antagonist, 8-phenyltheophylline (8-PT). The responses to ATP were little affected by 8-PT, indicating that ATP was not acting through breakdown to adenosine. At basal tone, ATP produced transient vasoconstriction only at concentrations greater than 0.1 mm, reaching a maximum of 38% of the contraction to 30mM KCl. 4 We conclude that in the rabbit coronary artery both NA and ATP produce vasodilatation by a direct action on the smooth muscle.

show abstract

An assessment of the antagonistic activity of reactive blue 2 at P1- and P2-purinoceptors: supporting evidence fro purinergic innervation of the rabbit portal vein

Cited by 47 publications

References 15 publications

Reactive Blue 2 Induces Calcium Oscillations in HeLa Cells.

Reactive Blue 2 Induces Calcium Oscillations in HeLa Cells.

The use of antagonists to characterize the receptors mediating depolarization of the rat isolated vagus nerve by α,β‐methylene adenosine 5′‐triphosphate

Vasodilator response of coronary smooth muscle to the sympathetic co‐transmitters noradrenaline and adenosine 5′‐triphosphate

Contact Info

Product

Resources

About