Skip to main content
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1996 Mar;117(5):775–780. doi: 10.1111/j.1476-5381.1996.tb15259.x

Rise in cytosolic Ca2+ concentration induced by P2-purinoceptor activation in isolated myocytes from the rat gastrointestinal tract.

H M Blottière 1, G Loirand 1, P Pacaud 1
PMCID: PMC1909402  PMID: 8851489

Abstract

1. The changes in the free cytosolic Ca2+ concentration ([Ca2+]i) in response to agonists of P2-purinoceptors were studied in myocytes isolated from the longitudinal muscle layer of different regions of the rat gastrointestinal tract (stomach, jejunum, ileum, caecum and colon). [Ca2+]i was estimated by emission from the fluorescent dye, indo-1. 2. ATP and the P2Y-purinoceptor agonist, 2-methylthio-ATP (2-MeSATP), transiently increased [Ca2+]i in single myocytes from all segments of the gastrointestinal tract, whereas alpha,beta-methylene-ATP, a P2x-purinoceptor agonist, had no effect. 3. The rise in [Ca2+]i induced by ATP and 2-MeSATP was maintained in Ca(2+)-free solution but was abolished by depletion of the intracellular store with thapsigargin (1 microM). 4. Single myocytes from stomach, caecum and colon also responded to UTP by a transient increase in [Ca2+]i. 5. Individual myocytes responded to ATP, 2-McSATP and UTP in a nearly all-or-nothing manner. The increasing of agonist concentration enhanced the number of responding cells but did not increase the amplitude of the [Ca2+]i rise. 6. These results suggest that myocytes from the longitudinal layer of gastrointestinal muscle do not possess functional P2x-purinoceptors and that agonists of P2Y and P2U-purinoceptors induced a rise in [Ca2+]i, probably via an all-or-nothing mobilization of Ca2+ from intracellular stores.

Full text

PDF
775

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. Benham C. D., Tsien R. W. A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle. Nature. 1987 Jul 16;328(6127):275–278. doi: 10.1038/328275a0. [DOI] [PubMed] [Google Scholar]
  3. Brake A. J., Wagenbach M. J., Julius D. New structural motif for ligand-gated ion channels defined by an ionotropic ATP receptor. Nature. 1994 Oct 6;371(6497):519–523. doi: 10.1038/371519a0. [DOI] [PubMed] [Google Scholar]
  4. Burnstock G., Campbell G., Satchell D., Smythe A. Evidence that adenosine triphosphate or a related nucleotide is the transmitter substance released by non-adrenergic inhibitory nerves in the gut. Br J Pharmacol. 1970 Dec;40(4):668–688. doi: 10.1111/j.1476-5381.1970.tb10646.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Furukawa K., Nomoto T. Postnatal changes in response to adenosine and adenine nucleotides in rat duodenum. Br J Pharmacol. 1989 Aug;97(4):1111–1118. doi: 10.1111/j.1476-5381.1989.tb12568.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  9. Iino M., Yamazawa T., Miyashita Y., Endo M., Kasai H. Critical intracellular Ca2+ concentration for all-or-none Ca2+ spiking in single smooth muscle cells. EMBO J. 1993 Dec 15;12(13):5287–5291. doi: 10.1002/j.1460-2075.1993.tb06224.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Kennedy I., Humphrey P. P. Evidence for the presence of two types of P2 purinoceptor in the guinea-pig ileal longitudinal smooth muscle preparation. Eur J Pharmacol. 1994 Aug 22;261(3):273–280. doi: 10.1016/0014-2999(94)90117-1. [DOI] [PubMed] [Google Scholar]
  13. Loirand G., Pacaud P. Mechanism of the ATP-induced rise in cytosolic Ca2+ in freshly isolated smooth muscle cells from human saphenous vein. Pflugers Arch. 1995 Jul;430(3):429–436. doi: 10.1007/BF00373919. [DOI] [PubMed] [Google Scholar]
  14. Manzini S., Maggi C. A., Meli A. Further evidence for involvement of adenosine-5'-triphosphate in non-adrenergic non-cholinergic relaxation of the isolated rat duodenum. Eur J Pharmacol. 1985 Jul 31;113(3):399–408. doi: 10.1016/0014-2999(85)90088-3. [DOI] [PubMed] [Google Scholar]
  15. Matharu M. S., Hollingsworth M. Purinoceptors mediating relaxation and spasm in the rat gastric fundus. Br J Pharmacol. 1992 Jun;106(2):395–403. doi: 10.1111/j.1476-5381.1992.tb14346.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moody C. J., Burnstock G. Evidence for the presence of P1-purinoceptors on cholinergic nerve terminals in the guinea-pig ileum. Eur J Pharmacol. 1982 Jan 8;77(1):1–9. doi: 10.1016/0014-2999(82)90527-1. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Ohta T., Ito S., Nakazato Y. All-or-nothing responses to carbachol in single intestinal smooth muscle cells of rat. Br J Pharmacol. 1994 Jul;112(3):972–976. doi: 10.1111/j.1476-5381.1994.tb13176.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pacaud P., Bolton T. B. Relation between muscarinic receptor cationic current and internal calcium in guinea-pig jejunal smooth muscle cells. J Physiol. 1991 Sep;441:477–499. doi: 10.1113/jphysiol.1991.sp018763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pacaud P., Grégoire G., Loirand G. Release of Ca2+ from intracellular store in smooth muscle cells of rat portal vein by ATP-induced Ca2+ entry. Br J Pharmacol. 1994 Oct;113(2):457–462. doi: 10.1111/j.1476-5381.1994.tb17011.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pacaud P., Loirand G., Baron A., Mironneau C., Mironneau J. Ca2+ channel activation and membrane depolarization mediated by Cl- channels in response to noradrenaline in vascular myocytes. Br J Pharmacol. 1991 Dec;104(4):1000–1006. doi: 10.1111/j.1476-5381.1991.tb12540.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pacaud P., Loirand G., Grégoire G., Mironneau C., Mironneau J. Noradrenaline-activated heparin-sensitive Ca2+ entry after depletion of intracellular Ca2+ store in portal vein smooth muscle cells. J Biol Chem. 1993 Feb 25;268(6):3866–3872. [PubMed] [Google Scholar]
  23. Pacaud P., Malam-Souley R., Loirand G., Desgranges C. ATP raises [Ca2+]i via different P2-receptor subtypes in freshly isolated and cultured aortic myocytes. Am J Physiol. 1995 Jul;269(1 Pt 2):H30–H36. doi: 10.1152/ajpheart.1995.269.1.H30. [DOI] [PubMed] [Google Scholar]
  24. Satchell D. The effects of ATP and related nucleotides on visceral smooth muscle. Ann N Y Acad Sci. 1990;603:53–63. doi: 10.1111/j.1749-6632.1990.tb37661.x. [DOI] [PubMed] [Google Scholar]
  25. Valera S., Hussy N., Evans R. J., Adami N., North R. A., Surprenant A., Buell G. A new class of ligand-gated ion channel defined by P2x receptor for extracellular ATP. Nature. 1994 Oct 6;371(6497):516–519. doi: 10.1038/371516a0. [DOI] [PubMed] [Google Scholar]
  26. Westfall D. P., Sedaa K. O., Shinozuka K., Bjur R. A., Buxton I. L. ATP as a cotransmitter. Ann N Y Acad Sci. 1990;603:300–310. doi: 10.1111/j.1749-6632.1990.tb37681.x. [DOI] [PubMed] [Google Scholar]
  27. Wiklund N. P., Gustafsson L. E. Indications for P2-purinoceptor subtypes in guinea pig smooth muscle. Eur J Pharmacol. 1988 Apr 13;148(3):361–370. doi: 10.1016/0014-2999(88)90114-8. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES