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. 1996 Mar;117(5):823–830. doi: 10.1111/j.1476-5381.1996.tb15267.x

Responses of the longitudinal muscle and the muscularis mucosae of the rat duodenum to adenine and uracil nucleotides.

C R Johnson 1, S J Charlton 1, S M Hourani 1
PMCID: PMC1909395  PMID: 8851497

Abstract

1. Previous studies have shown that the rat duodenum contains P1 and P2Y purinoceptors via which it relaxes to adenosine and adenosine 5'-triphosphate (ATP) respectively. It has also been shown to contract to uridine 5'-triphosphate (UTP) and adenosine 5'-O-(3-thiotriphosphate) (ATP-gamma-S), and based on their differential inhibition by the P2 antagonist suramin it has been suggested that they act via two separate receptors. In addition, the rat duodenum has been shown to dephosphorylate ATP rapidly via ectonucleotidases and adenosine deaminase. In this study the responses of two preparations from the rat duodenum, the longitudinal muscle and the muscularis mucosae, were investigated using a series of nucleotides and suramin. 2. 2-Methylthioadenosine 5'-triphosphate (2-MeSATP), ATP, ATP-gamma-S and adenosine 5'-alpha,beta-methylene-triphosphonate (AMPCPP) each relaxed the longitudinal muscle, with an agonist potency order of 2-MeSATP > ATP = ATP-gamma-S > AMPCPP, while UTP and uridine 5'-diphosphate (UDP) were not observed to elicit relaxation. This indicates the presence of a relaxant P2Y-purinoceptor on the longitudinal muscle. The longitudinal muscle did not contract to any of the agonists at concentrations of 300 microM, apart from ATP-gamma-S which caused very weak contractions. 3. ATP-gamma-S, adenosine 5'-methylenediphosphonate (AMPCP), AMPCPP, ATP, UTP, adenosine 5'-diphosphate (ADP), UDP and 2-MeSATP each contracted the muscularis mucosae with an agonist potency order of ATP-gamma-S > or = AMPCP > or = AMPCPP = ATP = UTP = ADP = UDP >> 2-MeSATP, although maximal responses were not obtained at concentrations of 300 microM. The muscularis mucosae did not relax to any of the agonists at concentrations of 300 microM. 4. Suramin (1 mM) inhibited relaxations induced by ATP on the longitudinal muscle, shifting the relaxation concentration-response curve to the right. This further supports the presence of a P2Y-purinoceptor on this muscle layer. Suramin (1 mM) inhibited contractions induced by AMPCPP, but not those induced by ATP, UTP or ATP-gamma-S, in the muscularis mucosae. Desensitization of the muscularis mucosae was seen with AMPCPP, but not with UTP or ATP-gamma-S, and no cross-desensitization between AMPCPP and UTP or ATP-gamma-S was observed. This suggests there are two receptors which mediate contraction on the rat duodenum muscularis mucosae, one suramin-sensitive and the other suramin-insensitive. 5. ATP was rapidly degraded by the muscularis mucosae to ADP, adenosine 5'-monophosphate (AMP) and inosine, with no adenosine being detected. A similar rate of degradation was seen for UTP with UDP, uridine 5'-monophosphate (UMP) and uridine being formed and for 2-MeSATP with 2-methylthioadenosine 5'-diphosphate (2-MeSADP), 2-methylthioadenosine 5'-monophosphate (2-MeSAMP) and 2-methylthioadenosine being formed. AMPCPP and ATP-gamma-S were both degraded more slowly, AMPCPP being degraded to AMPCP, and ATP-gamma-S to ADP, AMP and inosine. Suramin (1 mM), did not significantly affect the rate and pattern of degradation of these nucleotides, apart from AMPCPP which was degraded slightly more slowly in the presence of suramin. 6. These results show that there is a P2Y-purinoceptor which mediates relaxation in the rat duodenum longitudinal muscle. They also show that there is a contraction-mediating suramin-sensitive receptor on the rat duodenum muscularis mucosae which is desensitized by AMPCPP, and thus is probably of the P2X subtype. In addition, there is a contraction-mediating suramin-insensitive receptor on the rat duodenum muscularis mucosae which is not desensitized by UTP or ATP-gamma-S, and at which ATP and UTP show equal potency, and is thus probably of the P2U subtype. In addition, the rat duodenum muscularis mucosae contains ectonucleotidases and adenosine deaminase, which rapidly degrade nucleotides, although the inhibition by suramin of this deg

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Selected References

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  1. Bailey S. J., Hickman D., Hourani S. M. Characterization of the P1-purinoceptors mediating contraction of the rat colon muscularis mucosae. Br J Pharmacol. 1992 Feb;105(2):400–404. doi: 10.1111/j.1476-5381.1992.tb14265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bailey S. J., Hourani S. M. A study of the purinoceptors mediating contraction in the rat colon. Br J Pharmacol. 1990 Aug;100(4):753–756. doi: 10.1111/j.1476-5381.1990.tb14087.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailey S. J., Hourani S. M. Effects of purines on the longitudinal muscle of the rat colon. Br J Pharmacol. 1992 Apr;105(4):885–892. doi: 10.1111/j.1476-5381.1992.tb09073.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burnstock G., Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? Gen Pharmacol. 1985;16(5):433–440. doi: 10.1016/0306-3623(85)90001-1. [DOI] [PubMed] [Google Scholar]
  5. Collis M. G., Hourani S. M. Adenosine receptor subtypes. Trends Pharmacol Sci. 1993 Oct;14(10):360–366. doi: 10.1016/0165-6147(93)90094-z. [DOI] [PubMed] [Google Scholar]
  6. Connolly G. P. Evidence from desensitization studies for distinct receptors for ATP and UTP on the rat superior cervical ganglion. Br J Pharmacol. 1994 Jun;112(2):357–359. doi: 10.1111/j.1476-5381.1994.tb13079.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crack B. E., Beukers M. W., McKechnie K. C., Ijzerman A. P., Leff P. Pharmacological analysis of ecto-ATPase inhibition: evidence for combined enzyme inhibition and receptor antagonism in P2X-purinoceptor ligands. Br J Pharmacol. 1994 Dec;113(4):1432–1438. doi: 10.1111/j.1476-5381.1994.tb17157.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crack B. E., Pollard C. E., Beukers M. W., Roberts S. M., Hunt S. F., Ingall A. H., McKechnie K. C., IJzerman A. P., Leff P. Pharmacological and biochemical analysis of FPL 67156, a novel, selective inhibitor of ecto-ATPase. Br J Pharmacol. 1995 Jan;114(2):475–481. doi: 10.1111/j.1476-5381.1995.tb13251.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubyak G. R., el-Moatassim C. Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol. 1993 Sep;265(3 Pt 1):C577–C606. doi: 10.1152/ajpcell.1993.265.3.C577. [DOI] [PubMed] [Google Scholar]
  10. Filtz T. M., Li Q., Boyer J. L., Nicholas R. A., Harden T. K. Expression of a cloned P2Y purinergic receptor that couples to phospholipase C. Mol Pharmacol. 1994 Jul;46(1):8–14. [PubMed] [Google Scholar]
  11. Franco R., Hoyle C. H., Centelles J. J., Burnstock G. Degradation of adenosine by extracellular adenosine deaminase in the rat duodenum. Gen Pharmacol. 1988;19(5):679–681. doi: 10.1016/0306-3623(88)90128-0. [DOI] [PubMed] [Google Scholar]
  12. Fredholm B. B., Abbracchio M. P., Burnstock G., Daly J. W., Harden T. K., Jacobson K. A., Leff P., Williams M. Nomenclature and classification of purinoceptors. Pharmacol Rev. 1994 Jun;46(2):143–156. [PMC free article] [PubMed] [Google Scholar]
  13. Henderson D. J., Elliot D. G., Smith G. M., Webb T. E., Dainty I. A. Cloning and characterisation of a bovine P2Y receptor. Biochem Biophys Res Commun. 1995 Jul 17;212(2):648–656. doi: 10.1006/bbrc.1995.2018. [DOI] [PubMed] [Google Scholar]
  14. Hourani S. M., Chown J. A. The effects of some possible inhibitors of ectonucleotidases on the breakdown and pharmacological effects of ATP in the guinea-pig urinary bladder. Gen Pharmacol. 1989;20(4):413–416. doi: 10.1016/0306-3623(89)90188-2. [DOI] [PubMed] [Google Scholar]
  15. Hourani S. M., Johnson C. R., Bailey S. J. Desensitization of the P2-purinoceptors on the rat colon muscularis mucosae. Br J Pharmacol. 1993 Sep;110(1):501–505. doi: 10.1111/j.1476-5381.1993.tb13839.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hoyle C. H., Knight G. E., Burnstock G. Suramin antagonizes responses to P2-purinoceptor agonists and purinergic nerve stimulation in the guinea-pig urinary bladder and taenia coli. Br J Pharmacol. 1990 Mar;99(3):617–621. doi: 10.1111/j.1476-5381.1990.tb12979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johnson C. R., Hourani S. M. Contractile effects of uridine 5'-triphosphate in the rat duodenum. Br J Pharmacol. 1994 Dec;113(4):1191–1196. doi: 10.1111/j.1476-5381.1994.tb17123.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kasakov L., Burnstock G. The use of the slowly degradable analog, alpha, beta-methylene ATP, to produce desensitisation of the P2-purinoceptor: effect on non-adrenergic, non-cholinergic responses of the guinea-pig urinary bladder. Eur J Pharmacol. 1982 Dec 24;86(2):291–294. doi: 10.1016/0014-2999(82)90330-2. [DOI] [PubMed] [Google Scholar]
  19. Kennedy C., Leff P. How should P2X purinoceptors be classified pharmacologically? Trends Pharmacol Sci. 1995 May;16(5):168–174. doi: 10.1016/s0165-6147(00)89010-0. [DOI] [PubMed] [Google Scholar]
  20. Lazarowski E. R., Harden T. K. Identification of a uridine nucleotide-selective G-protein-linked receptor that activates phospholipase C. J Biol Chem. 1994 Apr 22;269(16):11830–11836. [PubMed] [Google Scholar]
  21. Leff P., Wood B. E., O'Connor S. E. Suramin is a slowly-equilibrating but competitive antagonist at P2x-receptors in the rabbit isolated ear artery. Br J Pharmacol. 1990 Nov;101(3):645–649. doi: 10.1111/j.1476-5381.1990.tb14134.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Meldrum L. A., Burnstock G. Evidence that ATP acts as a co-transmitter with noradrenaline in sympathetic nerves supplying the guinea-pig vas deferens. Eur J Pharmacol. 1983 Aug 19;92(1-2):161–163. doi: 10.1016/0014-2999(83)90126-7. [DOI] [PubMed] [Google Scholar]
  23. Murrin R. J., Boarder M. R. Neuronal "nucleotide" receptor linked to phospholipase C and phospholipase D? Stimulation of PC12 cells by ATP analogues and UTP. Mol Pharmacol. 1992 Mar;41(3):561–568. [PubMed] [Google Scholar]
  24. Nicholls J., Brownhill V. R., Hourani S. M. Characterization of P1-purinoceptors on rat isolated duodenum longitudinal muscle and muscularis mucosae. Br J Pharmacol. 1996 Jan;117(1):170–174. doi: 10.1111/j.1476-5381.1996.tb15170.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nicholls J., Hourani S. M., Kitchen I. Characterization of P1-purinoceptors on rat duodenum and urinary bladder. Br J Pharmacol. 1992 Mar;105(3):639–642. doi: 10.1111/j.1476-5381.1992.tb09032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nicholls J., Hourani S. M., Kitchen I. The ontogeny of purinoceptors in rat urinary bladder and duodenum. Br J Pharmacol. 1990 Aug;100(4):874–878. doi: 10.1111/j.1476-5381.1990.tb14107.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Connor S. E., Dainty I. A., Leff P. Further subclassification of ATP receptors based on agonist studies. Trends Pharmacol Sci. 1991 Apr;12(4):137–141. doi: 10.1016/0165-6147(91)90530-6. [DOI] [PubMed] [Google Scholar]
  28. O'Connor S. E. Recent developments in the classification and functional significance of receptors for ATP and UTP, evidence for nucleotide receptors. Life Sci. 1992;50(22):1657–1664. doi: 10.1016/0024-3205(92)90420-t. [DOI] [PubMed] [Google Scholar]
  29. Satchell D. G., Maguire M. H. Inhibitory effects of adenine nucleotide analogs on the isolated guinea-pig taenia coli. J Pharmacol Exp Ther. 1975 Dec;195(3):540–548. [PubMed] [Google Scholar]
  30. Saïag B., Milon D., Allaín H., Rault B., Van den Driessche J. Constriction of the smooth muscle of rat tail and femoral arteries and dog saphenous vein is induced by uridine triphosphate via 'pyrimidinoceptors', and by adenosine triphosphate via P2x purinoceptors. Blood Vessels. 1990;27(6):352–364. doi: 10.1159/000158829. [DOI] [PubMed] [Google Scholar]
  31. Seifert R., Schultz G. Involvement of pyrimidinoceptors in the regulation of cell functions by uridine and by uracil nucleotides. Trends Pharmacol Sci. 1989 Sep;10(9):365–369. doi: 10.1016/0165-6147(89)90009-6. [DOI] [PubMed] [Google Scholar]
  32. Surprenant A., Buell G., North R. A. P2X receptors bring new structure to ligand-gated ion channels. Trends Neurosci. 1995 May;18(5):224–229. doi: 10.1016/0166-2236(95)93907-f. [DOI] [PubMed] [Google Scholar]
  33. Tokuyama Y., Hara M., Jones E. M., Fan Z., Bell G. I. Cloning of rat and mouse P2Y purinoceptors. Biochem Biophys Res Commun. 1995 Jun 6;211(1):211–218. doi: 10.1006/bbrc.1995.1798. [DOI] [PubMed] [Google Scholar]
  34. Webb T. E., Simon J., Krishek B. J., Bateson A. N., Smart T. G., King B. F., Burnstock G., Barnard E. A. Cloning and functional expression of a brain G-protein-coupled ATP receptor. FEBS Lett. 1993 Jun 14;324(2):219–225. doi: 10.1016/0014-5793(93)81397-i. [DOI] [PubMed] [Google Scholar]
  35. Welford L. A., Cusack N. J., Hourani S. M. ATP analogues and the guinea-pig taenia coli: a comparison of the structure-activity relationships of ectonucleotidases with those of the P2-purinoceptor. Eur J Pharmacol. 1986 Oct 7;129(3):217–224. doi: 10.1016/0014-2999(86)90431-0. [DOI] [PubMed] [Google Scholar]
  36. Welford L. A., Cusack N. J., Hourani S. M. The structure-activity relationships of ectonucleotidases and of excitatory P2-purinoceptors: evidence that dephosphorylation of ATP analogues reduces pharmacological potency. Eur J Pharmacol. 1987 Sep 2;141(1):123–130. doi: 10.1016/0014-2999(87)90418-3. [DOI] [PubMed] [Google Scholar]
  37. Wilkinson G. F., McKechnie K., Dainty I. A., Boarder M. R. P2Y purinoceptor and nucleotide receptor-induced relaxation of precontracted bovine aortic collateral artery rings: differential sensitivity to suramin and indomethacin. J Pharmacol Exp Ther. 1994 Feb;268(2):881–887. [PubMed] [Google Scholar]
  38. Wilkinson G. F., Purkiss J. R., Boarder M. R. Differential heterologous and homologous desensitization of two receptors for ATP (P2y purinoceptors and nucleotide receptors) coexisting on endothelial cells. Mol Pharmacol. 1994 Apr;45(4):731–736. [PubMed] [Google Scholar]
  39. Wilkinson G. F., Purkiss J. R., Boarder M. R. The regulation of aortic endothelial cells by purines and pyrimidines involves co-existing P2y-purinoceptors and nucleotide receptors linked to phospholipase C. Br J Pharmacol. 1993 Mar;108(3):689–693. doi: 10.1111/j.1476-5381.1993.tb12862.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Windscheif U., Pfaff O., Ziganshin A. U., Hoyle C. H., Bäumert H. G., Mutschler E., Burnstock G., Lambrecht G. Inhibitory action of PPADS on relaxant responses to adenine nucleotides or electrical field stimulation in guinea-pig taenia coli and rat duodenum. Br J Pharmacol. 1995 Aug;115(8):1509–1517. doi: 10.1111/j.1476-5381.1995.tb16644.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Kügelgen I., Bültmann R., Starke K. Interaction of adenine nucleotides, UTP and suramin in mouse vas deferens: suramin-sensitive and suramin-insensitive components in the contractile effect of ATP. Naunyn Schmiedebergs Arch Pharmacol. 1990 Aug;342(2):198–205. doi: 10.1007/BF00166965. [DOI] [PubMed] [Google Scholar]
  42. von Kügelgen I., Häussinger D., Starke K. Evidence for a vasoconstriction-mediating receptor for UTP, distinct from the P2 purinoceptor, in rabbit ear artery. Naunyn Schmiedebergs Arch Pharmacol. 1987 Nov;336(5):556–560. doi: 10.1007/BF00169313. [DOI] [PubMed] [Google Scholar]
  43. von Kügelgen I., Starke K. Evidence for two separate vasoconstriction-mediating nucleotide receptors, both distinct from the P2x-receptor, in rabbit basilar artery: a receptor for pyrimidine nucleotides and a receptor for purine nucleotides. Naunyn Schmiedebergs Arch Pharmacol. 1990 Jun;341(6):538–546. doi: 10.1007/BF00171734. [DOI] [PubMed] [Google Scholar]

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