Biomedical subjects

S M Hourani

Publications and source records attributed to S M Hourani.

At least 19 recordsLinked to original sources

PubMed2001

Role of cyclic nucleotides in vasodilations of the rat thoracic aorta induced by adenosine analogues.

Although adenosine analogues such as 5'-N-ethylcarboxamidoadenosine (NECA) relax the rat thoracic aorta in a partially endothelium-dependent manner via adenosine A(2A) receptors, others such as N(6)-R-phenylisopropyladenosine (R-PIA) act via an endothelium-independent, antagonist-insensitive mechanism. The role of cyclic nucleotides in these relaxations was investigated in isolated aortic rings using inhibitors of adenylate and guanylate cyclases as well as subtype-selective phosphodiesterase inhibitors. The adenylate cyclase inhibitor 9-(tetrahydro-2-furanyl)-9H-purin-6-amine (SQ 22536; 100 microM) significantly inhibited responses to NECA, but not responses to R-PIA. The type IV (cyclic AMP-selective) phosphodiesterase inhibitor 4-[(3-butoxy-4-methoxyphenyl)methyl]-2-imidazolidinone (RO 20-1724; 30 microM) significantly enhanced responses to NECA and to a lesser extent those to R-PIA. The guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3a]quinoxalin-1-one (ODQ; 100 microM) significantly inhibited responses to NECA and acetylcholine but not responses to R-PIA. The selective phosphodiesterase V (cyclic GMP-selective) inhibitors, zaprinast (10 microM) and 4-[[3',4'-(methylenedioxy)benzyl]amino]-6-methoxyquinazoline (MMQ; 1 microM), had no significant effect on responses to either NECA or R-PIA, but enhanced responses to acetylcholine. These results are consistent with the effects of NECA being via activation of endothelial receptors to release NO which stimulates guanylate cyclase, as well as smooth muscle receptors coupled to stimulation of adenylate cyclase. The lack of effect of zaprinast and MMQ on responses to NECA are likely to be due to simultaneous activation of both adenylate and guanylate cyclases in the smooth muscle, as cyclic AMP reduces the sensitivity of phosphodiesterase V to inhibitors. These results also suggest that the effects of R-PIA are via neither of these mechanisms.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗

PubMed2000

Platelet P2 receptors: from curiosity to clinical targets.

Adenosine 5'-diphosphate (ADP) is a paracrine mediator that activates human blood platelets, causing them to become adhesive and thereby contributing to their role in hemostasis. The actions of ADP were initially thought to be mediated by a unique ADP receptor termed P2(T) found only on platelets and antagonized by ATP, but it appears that at least two P2Y receptor subtypes are involved, a P2Y(1) receptor linked in some way to control of intracellular-free calcium levels and another P2Y receptor linked via an inhibitory G protein to adenylate cyclase. In addition, the presence of excitatory P2X(1) receptors that mediate the influx of monovalent and divalent cations in response to both ADP and ATP has been demonstrated. The precise contribution that each of these P2 receptors make to the overall phenomena associated with platelet aggregation, adhesion and hemostasis is yet to be defined. Antithrombotic agents that interfere with the actions of ADP are marketed, and P2 receptor antagonists are entering clinical trials for acute treatments of thrombosis. This review seeks to summarize the present state of knowledge of platelet P2 receptor pharmacology and therapeutics.

Adenosine Diphosphate↗

PubMed2000

Breakdown of extracellular ATP by the prostatic and epididymal ends of the guinea pig vas deferens.

Adenosine 5'-triphosphate (ATP) has a higher potency at the prostatic than at the epididymal end of the guinea pig vas deferens. We bisected the tissue and measured the breakdown of ATP by each half, and although the half-lives differed, the rate constants per gram of tissue were not significantly different. For a range of tissue portions, a correlation was found between the portion weight and the half-life of ATP. The difference in half-life at the two ends is therefore due to the different weights of the tissues, and the difference in potency of ATP cannot be explained by differences in degradation.

Adenosine↗

PubMed2000

Characterisation of adenosine receptors mediating relaxation in hamster isolated aorta.

The aim of this study was to characterise the receptor(s) mediating relaxations to adenosine and its analogues in the hamster isolated aorta. Adenosine relaxed the aorta but there was no significant difference between pIC20 values in the absence and presence of 8-sulphophenyltheophylline (8-SPT, 50 microM), although there was a small right-shift (approximately threefold) of the lower portion of the curve in the presence of 8-SPT. However, in the presence of the adenosine uptake inhibitor nitrobenzylthioinosine (NBTI, 1 microM), curves to adenosine were left-shifted by approximately 100-fold and an apparent pK(B) for 8-SPT of 5.79+/-0.05 was obtained. Likewise, 5'-N-ethylcarboxamidoadenosine (NECA) relaxed the aorta but curves were biphasic. The first phase of the curve was blocked by 8-SPT (10-100 microM, pA2 = 5.75+/-0.14) and the A2A-selective antagonist 4-(2-[7-amino-2-(2-furyl) [1,2,4]-triazolo[2,3-a][1,3,5]triazin-5-ylaminolethyl) phenol (ZM 241385, 3 nM-1 microM, pK(B)=9.17+/-0.10). Similarly, the A2A-selective agonist 2-[p)-(2-carbonylethyl)-phenylethylamino]-5'-N-ethylcarboxam idoadenosine (CGS 21680) relaxed the tissues but curves were biphasic and the first phase was again blocked by ZM 241385 (10 nM, apparent pK(B)=9.06+/-0.34). In contrast, relaxations to N6-R-phenylisopropyladenosine (R-PIA), N6-cyclopentyladenosine (CPA), 2-chloroadenosine (2-CADO) and N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide (IB-MECA) were not blocked by 8-SPT (50 microM). Responses to IB-MECA were also not blocked by the A3 receptor antagonist 3-ethyl-5-benzyl-2-methyl-6-phenyl-4-phenylethynyl-1,4-(+/-)-dihyd ropyridine-3,5-dicarboxylate (MRS 1191, 30 microM). The asymptote of the first phase of curves to NECA was markedly reduced (and in some preparations the first phase was completely abolished) both in the presence of N(G)-nitro-L-arginine methyl ester (L-NAME, 0.1 mM), and in the absence of endothelium. Likewise, the first phase of curves to CGS 21680 was abolished both in the presence of L-NAME (0.1 mM) and in the absence of endothelium. In contrast, there were only relatively small shifts to the right of curves to adenosine and the other analogues in the presence of L-NAME or the absence of endothelium (between three- and fivefold). The data suggest the presence of A2A receptors which are located on the endothelium and mediate release of nitric oxide. These receptors are activated by NECA, CGS 21680 and adenosine (in the presence of uptake blockade). The resistance to blockade of relaxations to adenosine (in the absence of uptake inhibitor), CPA, R-PIA, 2-CADO, IB-MECA and high concentrations of NECA and CGS 21680 by 8-SPT or ZM 241385 suggests the presence of an additional mechanism(s). Data obtained with adenosine in the absence and presence of NBTI suggest that the endogenous ligand may cause relaxation via an intracellular mechanism.

2-Chloroadenosine↗

PubMed1999

Ontogeny of adenosine receptors in the longitudinal muscle and muscularis mucosae of the rat distal colon.

The development of adenosine A1 and A2B receptors on the longitudinal muscle and muscularis mucosae of the neonatal rat distal colon has been investigated using homogenate binding, quantitative autoradiography and functional studies. In homogenate binding studies 1,3-[3H]-dipropyl-8-cyclopentylxanthine ([3H]DPCPX) bound with high affinity to A1 receptors in the muscularis mucosae and intact colon from rats aged 10, 15, 20, 25 and 30 days. The affinity of [3H]DPCPX was similar to that in the adult at all ages, but the density of binding sites was higher in the neonatal tissues. Quantitative autoradiography showed a higher density of [3H]DPCPX binding sites in the longitudinal muscle than in the muscularis mucosae at all ages studied (day 10 to adult), and this binding was concentration-dependently displaced by N6-cyclopentyladenosine (CPA). In functional studies the longitudinal muscle relaxed in response to 5'-N-ethylcarboxamidoadenosine (NECA) and CPA at all ages studied (15-30 days), NECA being more potent than CPA. The potency of NECA remained constant and it was antagonised by 1 microM DPCPX at all ages with pA2-values consistent with activation of A2 receptors. The inactivity of 2-[p-(carboxyethyl)-phenylethylamino]-5'-N-ethylcarboxamidoadenosi ne (CGS 21680) indicated that the A2 receptors were of the A2B subtype. The muscularis mucosae contracted in response to CPA at all ages studied (day 15 to adult) and the antagonism by DPCPX (10 nM) were consistent with activation of A1 receptors. In conclusion, binding, autoradiographic and functional studies identified A1 receptors on the rat colon muscularis mucosae at all ages studied. However, while binding and autoradiographic localisation showed the presence of A1 receptors in the longitudinal muscle at all ages studied, functional data only revealed the presence of A2B receptors.

Adenosine↗

PubMed1999

Postnatal development of purinoceptors in rat visceral smooth muscle preparations.

1. Adenosine and ATP have well-established functions as neuromodulator and neurotransmitter, respectively, in smooth muscle preparations, and purinergic control may be an early form of autonomic control in both evolution and ontogenesis. 2. This review describes the postnatal development of responses mediated by the various receptors for adenosine and for nucleotides in the rat duodenum, colon, urinary bladder and vas deferens and considers the implications that this development may have for the importance of purinergic control in neonates and adults.

Age Factors↗

PubMed1999

Differential effects of adenine nucleotide analogues on shape change and aggregation induced by adnosine 5-diphosphate (ADP) in human platelets.

Adenosine 5'-diphosphate (ADP) induces human blood platelets to aggregate and change shape, and it has been suggested that these two responses are mediated by more than one subtype of ADP receptor. The structure-activity relationships for several analogues of adenine nucleotides in causing aggregation and shape change were measured and compared in washed platelets using an aggregometer. ADP and its analogues 2-methylthioadenosine 5'-diphosphate (2-methylthio-ADP), adenosine 5'(alpha,beta-methylene)diphosphonate (AMPCP), S(P)-adenosine 5'-O-(1-thiodiphosphate) (AD-P alphaS) and adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS) were used as agonists. Adenosine 5'-triphosphate (ATP) and its analogues, P1, P5-diadenosine pentaphosphate (ApsA), adenosine (5'-(alpha,beta-methylene)triphosphonate (AMPCPP), 2-methylthioadenosine 5'-triphosphate (2-methylthio-ATP) and uridine 5'-triphosphate (UTP), as well as the trypanocidal drug suramin, were used as antagonists. In general, the structure-activity relationships for both responses were similar, but for some analogues differences were observed. ADPalphaS and ADPbetaS were much more potent agonists relative to ADP for shape change than for aggregation and indeed ADPalphaS antagonized ADP-induced aggregation with an apparent pK(B) value of 5.5+/-0.1. 2-Methylthio-ATP also had different effects in aggregation and shape change, being a much higher affinity antagonist of aggregation than of shape change with an apparent pK(B) value of 7.0+/-0.2 for aggregation and 5.2+/-0.2 for shape change. These results support the suggestion that these two responses are mediated by multiple ADP receptors on human platelets, and are consistent with shape change being mediated via one receptor (the P2Y1 receptor) with aggregation requiring the activation of two receptors (the P2Y1 and another P2Y receptor).

Adenine Nucleotides↗

PubMed1999

Effects of noradrenaline, the calcium ionophore A23187, forskolin, sodium nitroprusside and glibenclamide on the degradation of extracellular adenosine 5'-triphosphate by the rat isolated vas deferens.

1. The effects of noradrenaline (NA), the calcium ionophore A23187, forskolin, sodium nitroprusside (SNP) and the K+-channel blocker glibenclamide on the degradation by ectonucleotidases of extracellular adenosine 5'-triphosphate (ATP) were studied in the rat vas deferens. 2. ATP (100 microM) was rapidly broken down by the rat vas deferens with a half-life of 5.83 +/- 0.40 min via adenosine 5'-diphosphate (ADP) and adenosine 5'-monophosphate (AMP), with the final degradation product being inosine and with little adenosine being detected in the samples. 3. Preincubation for 1 h with NA (10 microM), A23187 (10 microM), or glibenclamide (100 microM) had no significant effect on the breakdown of ATP or the production of metabolites. However, both forskolin (10 microM) and SNP (1 microM) significantly increased the concentrations of AMP detected with time. In the case of SNP (1 microM) there was also a significant reduction in the rate of production of inosine, while in the case of forskolin (10 microM) there was a significant increase in the rate of removal of ATP. 4. These results suggest that preincubation with SNP may inhibit 5'-nucleotidase and so reduce the metabolism of AMP, while preincubation with forskolin may increase the activity of the ectonucleotidases responsible for production of AMP from ATP.

5'-Nucleotidase↗

PubMed1999

Effects of allopurinol, erythro-9-(2-hydroxy-3-nonyl)adenine and S-(4-nitrobenzyl)-6-thioinosine on the degradation of adenosine 5'-triphosphate in the rat colon muscularis mucosae.

The effects on ATP breakdown of some modulators of adenosine transport or metabolism were studied in the rat colon muscularis mucosae, a tissue which contracts to ATP and is thought to contain P2Y1 receptors. The compounds tested were the xanthine oxidase inhibitor allopurinol, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and the adenosine uptake blocker S-(4-nitrobenzyl)-6-thioinosine (NBTI). The degradation of adenosine 5'-triphosphate (ATP) (100 microM) and the appearance of metabolites was followed by high pressure liquid chromatography during incubation of isolated tissue preparations alone or in the presence of the drugs, following preincubation with the drugs for 1 h. In the absence of drugs ATP was rapidly degraded by the rat colon muscularis mucosae with a half-life of 6.1 +/- 0.7 min, the major breakdown product being inosine rather than adenosine. Allopurinol (1 microM) and NBTI (10 microM) had no effect on the rate of breakdown of ATP or on the pattern of metabolites produced. EHNA (1 or 10 microM) also had no effect on the half-life of ATP, but in the presence of EHNA (1 microM) the rate of production of inosine was significantly reduced and some adenosine was detected, while in the presence of 10 microm EHNA the production of inosine was abolished and adenosine became the final breakdown product. These results indicate that allopurinol (1 microM) and NBTI (10 microM) have no detectable effect on extracellular purine metabolism in this tissue, and that the build-up of adenosine produced by treatment with EHNA does not have a feedback effect on ATP breakdown.

Adenine↗

PubMed1998

Effects of adenosine 5'-triphosphate, uridine 5'-triphosphate, adenosine 5'-tetraphosphate and diadenosine polyphosphates in guinea-pig taenia caeci and rat colon muscularis mucosae.

The functional effects of adenosine 5'-triphosphate (ATP), uridine 5'-triphosphate (UTP), adenosine 5'-tetraphosphate (AP4) and the diadenosine polyphosphates P1,P3-diadenosine triphosphate (Ap3A), P1,P4-diadenosine tetraphosphate (Ap4A) and P1,P5-diadenosine pentaphosphate (Ap5A) were studied in two isolated smooth muscle preparations thought to contain P2Y (P2Y1) receptors, the guinea-pig taenia caeci (which relaxes to ATP) and the rat colon muscularis mucosae (which contracts to ATP). In addition, the breakdown of these compounds by the rat colon muscularis mucosae was investigated by high pressure liquid chromatography. In the guinea-pig taenia caeci all the purine nucleotides caused relaxation with a potency order of Ap3A=Ap4A> ATP>AP4=Ap5A, and these relaxations were antagonised by suramin with apparent pA2 values in the region of 5, consistent with activation of a P2Y1 receptor. In the rat colon muscularis mucosae the nucleotides caused contraction with a potency order of Ap3A = Ap4A>ATP=AP4 =Ap5A >UTP. However, while suramin (100 microM) inhibited responses to ATP and UTP at all concentrations of agonist, it only inhibited contractions induced by the higher concentrations of AP4, Ap3A and Ap4A and had little effect on contractions induced by Ap5A. A higher concentration of suramin (1 mM) enhanced contractions induced by ATP but greatly inhibited those induced by UTP and had no effect on responses to the other agonists. The A1 adenosine receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX; 10 nM) had no effect on responses to ATP or UTP but inhibited responses to Ap3A, Ap4A, Ap5A and AP4. A combination of suramin (1 mM) and DPCPX (10 nM) almost abolished responses to all the agonists. ATP and UTP were rapidly degraded by the rat colon muscularis mucosae while AP4, Ap3A, Ap4A and Ap5A were degraded more slowly, and the major product detected after breakdown of the purine nucleotides was inosine rather than adenosine. The breakdown of all the nucleotides was inhibited by suramin (1 mM), although this inhibition did not achieve statistical significance in the case of ATP. These results show that while the diadenosine polyphosphates appear to act as P2 agonists in the taenia caeci, in the rat colon muscularis mucosae their major action is via adenosine A1 receptors rather than via P2 receptors. In addition, although they are more stable than ATP or UTP, their action in this tissue is clearly affected by their degradation which complicates the effects of suramin.

Adenine Nucleotides↗

PubMed1998

The regulation of vascular function by P2 receptors: multiple sites and multiple receptors.

Although the effects of nucleotides in the cardiovascular system have been known for almost 70 years, it is only in the past few years that some of the P2 receptors at which they act have been cloned and characterized. It is now clear that the control of cardiovascular function by nucleotides is complex, involving multiple receptors and multiple effects in the different cell types of importance. In this review Mike Boarder and Susanna Hourani summarize the P2 receptors that are present in endothelial cells, platelets, smooth muscle and nerves, the signalling pathways that they activate and the responses that are produced. They also discuss the important role of nucleotides in the interactions between the different cell types, and the implications of this in vascular disease.

Adenosine Triphosphate↗

PubMed1997

Characterization of adenosine receptors on rat ileum, ileal longitudinal muscle and muscularis mucosae.

Adenosine receptors were studied in isolated rat ileum, ileal longitudinal muscle and muscularis mucosae, using a range of agonists and an antagonist. In the rat ileal longitudinal muscle adenosine receptor agonists relaxed the tissues. N6-cyclopentyladenosine (CPA) was more potent than 5'-N-ethylcarboxamidoadenosine (NECA) or adenosine and 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) (1 nM) gave a 5-fold parallel shift to the right of the concentration-response curves to both CPA and NECA corresponding to an apparent pA2 value of 9.6 suggesting that the agonists relax via adenosine A1 receptors. In the intact ileum adenosine receptor agonists also relaxed the tissue but NECA and CPA were equipotent. DPCPX (3 nM) however inhibited responses to both CPA and NECA with dose-ratios of 8 and 15.6, corresponding to pA2 values of 9.3 and 9.7, respectively. DPCPX (300 nM) gave a much greater shift to the right of the concentration-response curve to NECA with a dose-ratio of 769, corresponding to an apparent pA2 of 9.4. This suggests that the agonists are acting at adenosine A1 receptors to cause relaxation of the whole tissue. Adenosine receptor agonists contracted rat ileal muscularis mucosae with a potency order indicative of an A adenosine receptor. DPCPX (3-100 nM) antagonized responses to CPA giving a linear Schild plot with a slope close to unity and a pA2 of 8.4 suggesting an action on adenosine A1 receptors.

Adenosine↗

PubMed1997

Relaxation of the ovine isolated iris sphincter by adenosine receptor agonists: lack of effect of adenosine A1 and A2 receptor antagonists.

The effects of adenosine receptor ligands on the tone of the ovine isolated iris sphincter were investigated, and adenosine analogues were found to relax the carbachol-contracted tissue in a concentration-dependent manner with an order of potency of 5'-N-ethylcarboxamidoadenosine > or = 2-(p-(2-carboxyethyl)phenylethylamino)-5'-N-ethylcarboxamidoadenos ine (CGS 21680) > or = N6-cyclopentyladenosine > adenosine, consistent with activation of an adenosine A2A receptor. However, these responses were not inhibited by the non-selective adenosine A1/A2 receptor antagonist 8-p-sulphophenyltheophylline (50 microM), the selective adenosine A2A/A1 receptor antagonist N-[2-(dimethylamino)ethyl]-N-methyl-4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3- dipropyl-1H-purin-8-yl)benzenesulphonamide (PD 115,199) (0.1 microM) or the non-xanthine adenosine A2A receptor antagonist (4-(2-[7-amino-2-(2-furyl) [1,2,4]-triazolo[2,3-a][1,3,5]triazin-5-yl-amino]ethyl)phenol) (ZM 241385) (0.1 microM). The relaxations cannot therefore be mediated by activation of adenosine A1, A2A or A2B receptors.

Adenosine↗

PubMed1997

Adenosine analogues relax guinea-pig taenia caeci via an adenosine A2B receptor and a xanthine-resistant site.

In this study we have sub-classified the adenosine A2 receptor mediating relaxation in the guinea-pig taenia caecum using the adenosine A2A receptor-selective agonist CGS 21680 (2-[p-(2-carboxyethyl)phenylamino]-5'-N-ethylcarboxamidoadenosine) and the adenosine A2A receptor-selective antagonist ZM 241385 (4-(2-[7-amino-2-(2-furyl) [1,2,4]-triazolo[2,3-a][1,3,5]triazin-5-yl amino]ethyl)phenol). CGS 21680 did not elicit relaxations, and a pKB value of 7.80 was obtained for ZM 241385 against 5'-N-ethylcarboxamidoadenosine suggesting the presence of adenosine A2B receptors. Relaxations are also mediated via a xanthine-resistant site. In this study relaxations to the adenosine A3 receptor agonist IB-MECA (N6-(3-iodo-benzyl)adenosine-5'-N-methyluronamide) were blocked by neither 8-sulphophenyltheophylline (100 microM) nor the adenosine A3 receptor antagonist BW-A1433 (1,3-dipropyl-8-(4-acrylate)phenylxanthine, 100 microM), suggesting that this site is not an adenosine A3 receptor.

Adenosine↗

PubMed1997

Involvement of functional antagonism in the effects of adenosine antagonists and L-NAME in the rat isolated heart.

1. Adenosine and its analogues induced an increase in flow rate when infused into the rat isolated perfused heart. The agonist potency order obtained was 2-(rho-(2-carboxyethyl)phenethyl-amino)-5'-N-ethylcarboxamidoadenosine (CGS 21680) > or = 5'-N-ethylcarboxamidoadenosine (NECA) > N6-cyclopentyladenosine = adenosine, although the maximal response obtained for CGS 21680 was only 70% of that achieved by NECA. NG-Nitro-L-arginine methyl ester (L-NAME), the nonselective adenosine antagonist 8-rho-(sulfophenyl)theophylline (8-SPT) and the A2a selective antagonist N-[2-(dimethylamino)ethyl]-N-methyl-4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3- diprophyl-1H-purin-8-yl)benzene sulfonamide (PD 115, 119) reduced responses to the adenosine agonists, but some of this reduction was shown to be due to a nonspecific decrease in flow rate as well as a specific inhibitory action. 2. When this functional antagonism is taken into account, the results suggest that the increase in flow rate induced by the adenosine agonists was mediated by A2 receptors, with the increase in flow rate induced by CGS 21680 mediated by A2a receptors, whereas that induced by NECA was mediated by A2b receptors. 3. L-NAME did not appear to have any effect on the increase in flow rate induced by the adenosine agonists, suggesting that these responses were probably endothelium independent and do not involve the nitric oxide pathway.

Adenosine↗

PubMed1997

Ontogeny of P2-purinoceptors in the longitudinal muscle and muscularis mucosae of the rat isolated duodenum.

1. The ontogeny of P2-purinoceptors in the longitudinal muscle and the muscularis mucosae of the rat isolated duodenum was investigated by use of functional assays in tissues from neonatal animals. The degradation of purinoceptor agonists by the rat duodenum muscularis mucosae was also investigated. 2. In the rat duodenum muscularis mucosae adenosine 5'-(alpha, beta-methylene)triphosphonate (AMPCPP), adenosine 5'-triphosphate (ATP), uridine 5'-triphosphate (UTP) and 2-methylthioadenosine 5'-triphosphate (2-Me-S-ATP) all caused a contraction from day 10 to day 40, day 10 being the earliest age it could be tested. The potency order of agonists above day 25 was AMPCPP > ATP = UTP > 2-Me-S-ATP and this is similar to the potency order previously obtained for the adult tissue. However, in the neonatal tissues below day 20, 2-Me-S-ATP was the most potent agonist and at days 10 and 15 the order was 2-Me-S-ATP > AMPCPP > ATP = UTP. 3. In the rat duodenum muscularis mucosae desensitization was observed with AMPCPP at day 30 but not at day 15. At day 30, cross-desensitization was also observed between AMPCPP and 2-Me-S-ATP but not between AMPCPP and ATP or UTP, whereas no cross-desensitization was observed at day 15 with AMPCPP and any of the agonists. At day 15 and below AMPCPP and 2-Me-S-ATP may therefore both activate P2Y-receptors (2-Me-S-ATP > AMPCPP, no desensitization with AMPCPP) whereas above day 20 the agonists activate P2X-receptors (AMPCPP > 2-Me-S-ATP, desensitization with AMPCPP) which is similar to the adult tissue. Since ATP and UTP were equipotent in the muscularis mucosae and as no cross-desensitization was observed with AMPCPP and UTP or ATP at days 15 or 30, it is likely that ATP and UTP both activate P2U-receptors throughout the ages, as in the adult. 4. The potency of all the agonists in causing contraction in the rat duodenum muscularis mucosae decreased with age. The potency of AMPCPP and 2-Me-S-ATP in causing contractions was highest in the neonates before day 25, and reached values not significantly different from adult by day 30, and the potency of ATP and UTP causing contractions in this tissue was also highest in the neonates at days 10 and 15, and reached values not significantly different from adult by day 20. This suggests either that the receptor populations mediating contraction are highest in the neonates below day 20 or that the agonists are degraded by the muscularis mucosae to a greater extent after day 20. 5. In the rat duodenum muscularis mucosae the degradation of ATP, UTP, 2-Me-S-ATP and AMPCPP was followed by high pressure liquid chromatography at days 15 and 30. ATP was degraded to adenosine 5'-diphosphate (ADP), adenosine 5'-monophosphate (AMP) and inosine with no adenosine being detected, 2-Me-S-ATP was degraded to 2-methylthioadenosine 5'-diphosphate (2-Me-S-ADP), 2-methylthioadenosine 5'-monophosphate (2-Me-S-AMP) and 2-methylthioadenosine (2-Me-S-adenosine), and UTP was degraded to uridine 5'-diphosphate (UDP), uridine 5'-monophosphate (UMP) and uridine. The rate of degradation of these agonists was much faster at day 30 than at day 15, probably due to the increase in the size of the tissue. AMPCPP was also degraded with adenosine 5'-(alpha,beta-methylene)diphosphonate (AMPCP) being detected at both ages. However, at day 30 the rate of degradation of AMPCPP was much slower than for ATP, UTP or 2-Me-S-ATP. 6. In the rat duodenum longitudinal muscle 2-Me-S-ATP and AMPCPP both caused a relaxation with a potency order of 2-Me-S-ATP > AMPCPP, suggesting the activation of P2Y-receptors, as previously found for the adult tissue. Weak relaxations were observed to both the agonists at day 15 (the earliest age it could be studied), and the potency of the agonists reached values not significantly different from adult tissues by day 25. 7. Overall, these results suggest that in the neonatal rat duodenum longitudinal muscle there are P2Y-receptors mediating relaxation and that the receptor population i

Adenosine Triphosphate↗

PubMed1997

Activation of two sites by adenosine receptor agonists to cause relaxation in rat isolated mesenteric artery.

1. In this study we have characterized the receptor(s) in the rat mesenteric artery mediating relaxant responses to adenosine and a number of adenosine analogues, N6-R-phenylisopropyladenosine (R-PIA), N6-cyclopentyladenosine (CPA), N6-(3-iodo-benzyl)-adenosine-5'-N-methyluronamide (IB-MECA) and 5'-N-ethylcarboxamidoadenosine (NECA), by use of the non-selective antagonist 8-sulphophenyltheophylline (8-SPT) and the A2A selective ligands 2-[p-(2-carbonylethyl)-phenylethylamino]-5'-N-ethylcarboxami doadenosine (CGS 21680) and 4-(2-[7-amino-2-(2-furyl)[1,2,4]-triazolo[2,3-a][1,3,5]-triazin-5- ylamino]ethyl) phenol (ZM 241385). We have also studied the effects of endothelial removal and uptake inhibition by nitrobenzylthioinosine (NBTI) and the effects of the A3 receptor antagonist 1,3-dipropyl-8-(4-acrylate)phenylxanthine (BWA1433). 2. Adenosine, NECA, CPA and R-PIA all elicited relaxant responses in tissues precontracted with phenylephrine (1 microM) with the following potency order: NECA > R-PIA > adenosine = CPA. However, E/[A] curves to NECA were biphasic. CGS 21680 was inactive at concentrations up to 30 microM and IB-MECA elicited relaxant responses which were resistant to blockade by 8-SPT and BWA1433 (100 microM). 3. Removal of the endothelium produced a small but significant decrease in the asymptote of the high potency phase of E/[A] curves to NECA with no change in p[A]50. E/[A] curves to adenosine were not altered by removal of the endothelium. However, there were small rightward shifts of E/[A] curves to CPA and R-PIA in the absence of endothelium. 4. Inhibition of uptake by NBTI (1 microM) had no effect on E/[A] curves to NECA, CPA or R-PIA, but E/[A] curves to adenosine were significantly left-shifted in the presence of NBTI. 5. 8-SPT (10-100 microM) caused significant rightward shifts of the high potency phase of the E/[A] curves to NECA (pA2 = 5.63+/-0.26). The second phase of the concentration-response curve to NECA appeared to be resistant to blockade by 8-SPT, as were E/[A] curves for adenosine, CPA or R-PIA. However, in the presence of NBTI (1 microM), 8-SPT (100 microM) gave significant rightward shifts of E/[A] curves to adenosine. 6. ZM 241385 (0.1-1 microM) produced significant rightward shifts of the high potency phase of NECA E/[A] curves (pA2=7.65+/-0.25 in the presence and 7.20+/-0.12 in the absence of endothelium), while curves to R-PIA were not significantly shifted by 1 microM ZM 241385. In the presence of NBTI E/[A] curves to adenosine were significantly rightward shifted by ZM 241385 (0.1 microM, pA2=7.50+/-0.16). 7. In conclusion, the results suggest activation of A2B receptors located primarily on the smooth muscle by low concentrations of NECA and by adenosine under conditions of uptake blockade, and of another, as yet undefined site which may be intracellular, by higher concentrations of NECA, by CPA, R-PIA and adenosine under conditions where uptake is operational.

Adenosine↗

PubMed1997

Use of a newly developed technique to isolate rat pinealocytes and study the effects of adenosine agonists on melatonin production.

Recent studies have suggested a role for adenosine in the regulation of rat pineal melatonin synthesis. The data, however, are conflicting and therefore the aim of this study was to characterize adenosine receptors more fully in vitro by using a range of selective adenosine agonists and the adenosine antagonist 8-sulphophenyltheophylline (8-SPT). A simple method for the mechanical separation of rat pinealocytes was developed. Pinealocytes were briefly (15 min) incubated with drugs followed by a 4 hr drug-free incubation period after which melatonin concentrations in the incubation medium were measured by radioimmunoassay. The beta-adrenoceptor agonist isoprenaline gave a dose-related increase in melatonin production, demonstrating that this pinealocyte preparation technique is suitable to evaluate the effect of drugs on pineal melatonin synthesis. Our results show that adenosine, N6-(phenylisopropyl)adenosine (R-PIA) and 2-p-(2-carboxethyl) phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS21680) did not affect melatonin synthesis alone or in combination with isoprenaline. However 5'-N-ethylcarboxamidoadenosine (NECA) (100 microM) potentiated the stimulatory effect of isoprenaline (3 microM) on pineal melatonin production and this effect appeared to be antagonized by 8-SPT (50 microM). These results are consistent with activation by NECA of an A2b adenosine receptor.

Adenosine↗