Skip to main content
NCBI home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1992 Dec;107(4):1083–1087. doi: 10.1111/j.1476-5381.1992.tb13410.x

Inositol 1,4,5-trisphosphate generation and calcium mobilisation via activation of an atypical P2 receptor in the neuronal cell line, N1E-115.

P A Iredale 1, K F Martin 1, S P Alexander 1, S J Hill 1, D A Kendall 1
PMCID: PMC1907950  PMID: 1467830

Abstract

1. Alterations in the levels of intracellular calcium ([Ca2+]i) and D-myo-inositol-1,4,5-trisphosphate (InsP3) were measured in the murine neuroblastoma cell line clone, N1E-115, by use of the calcium-sensitive dye, fura-2 and a radioreceptor assay, respectively. 2. Exposure of the cells to ATP (100 microM) elicited rapid and transient increases in [Ca2+]i and InsP3, with both responses reaching a maximum between 10-20 s after agonist addition. 3. Investigation of concentration-response data by use of various analogues of ATP suggests the presence of an extracellular receptor which fails to fit into the current classification of purinoceptors. 4. Cross-desensitization experiments suggest that the same receptor can also be activated by the structurally different pyrimidine base, UTP. 5. Application of the tumour-promoting agent, beta-phorbol-12,13 dibutyrate (PDBu) caused a reduction in the increases in both [Ca2+]i and InsP3, suggesting a role for protein kinase C in feedback inhibition of purinoceptor responses in this cell line. 6. In summary, we present the first evidence for the existence of an atypical purinoceptor on a cell line of CNS origin. This receptor is linked to stimulation of phosphoinositide turnover and subsequent mobilisation of intracellular calcium.

Full text

PDF
1083

Selected References

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

  1. Allsup D. J., Boarder M. R. Comparison of P2 purinergic receptors of aortic endothelial cells with those of adrenal medulla: evidence for heterogeneity of receptor subtype and of inositol phosphate response. Mol Pharmacol. 1990 Jul;38(1):84–91. [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Brown H. A., Lazarowski E. R., Boucher R. C., Harden T. K. Evidence that UTP and ATP regulate phospholipase C through a common extracellular 5'-nucleotide receptor in human airway epithelial cells. Mol Pharmacol. 1991 Nov;40(5):648–655. [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. Burnstock G. Neural nomenclature. Nature. 1971 Jan 22;229(5282):282–283. doi: 10.1038/229282d0. [DOI] [PubMed] [Google Scholar]
  6. Challiss R. A., Batty I. H., Nahorski S. R. Mass measurements of inositol(1,4,5)trisphosphate in rat cerebral cortex slices using a radioreceptor assay: effects of neurotransmitters and depolarization. Biochem Biophys Res Commun. 1988 Dec 15;157(2):684–691. doi: 10.1016/s0006-291x(88)80304-8. [DOI] [PubMed] [Google Scholar]
  7. Cockcroft S., Gomperts B. D. The ATP4- receptor of rat mast cells. Biochem J. 1980 Jun 15;188(3):789–798. doi: 10.1042/bj1880789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cowen D. S., Sanders M., Dubyak G. P2-purinergic receptors activate a guanine nucleotide-dependent phospholipase C in membranes from HL-60 cells. Biochim Biophys Acta. 1990 Jul 12;1053(2-3):195–203. doi: 10.1016/0167-4889(90)90014-5. [DOI] [PubMed] [Google Scholar]
  9. Ehrlich Y. H., Snider R. M., Kornecki E., Garfield M. G., Lenox R. H. Modulation of neuronal signal transduction systems by extracellular ATP. J Neurochem. 1988 Jan;50(1):295–301. doi: 10.1111/j.1471-4159.1988.tb13263.x. [DOI] [PubMed] [Google Scholar]
  10. Fernández-Gallardo S., Gijón M. A., García C., Furio V., Liu F. T., Sánchez Crespo M. The role of platelet-activating factor and peptidoleukotrienes in the vascular changes of rat passive anaphylaxis. Br J Pharmacol. 1992 Jan;105(1):119–125. doi: 10.1111/j.1476-5381.1992.tb14221.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fine J., Cole P., Davidson J. S. Extracellular nucleotides stimulate receptor-mediated calcium mobilization and inositol phosphate production in human fibroblasts. Biochem J. 1989 Oct 15;263(2):371–376. doi: 10.1042/bj2630371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Forsberg E. J., Feuerstein G., Shohami E., Pollard H. B. Adenosine triphosphate stimulates inositol phospholipid metabolism and prostacyclin formation in adrenal medullary endothelial cells by means of P2-purinergic receptors. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5630–5634. doi: 10.1073/pnas.84.16.5630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gordon J. L. Extracellular ATP: effects, sources and fate. Biochem J. 1986 Jan 15;233(2):309–319. doi: 10.1042/bj2330309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Hallam T. J., Pearson J. D. Exogenous ATP raises cytoplasmic free calcium in fura-2 loaded piglet aortic endothelial cells. FEBS Lett. 1986 Oct 20;207(1):95–99. doi: 10.1016/0014-5793(86)80019-9. [DOI] [PubMed] [Google Scholar]
  16. Hoiting B., Molleman A., Nelemans A., Den Hertog A. P2-purinoceptor-activated membrane currents and inositol tetrakisphosphate formation are blocked by suramin. Eur J Pharmacol. 1990 May 31;181(1-2):127–131. doi: 10.1016/0014-2999(90)90253-3. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Iredale P. A., Martin K. F., Hill S. J., Kendall D. A. Agonist-induced changes in [Ca2+]i in N1E-115 cells: differential effects of bradykinin and carbachol. Eur J Pharmacol. 1992 Jun 5;226(2):163–168. doi: 10.1016/0922-4106(92)90178-x. [DOI] [PubMed] [Google Scholar]
  19. Iredale P. A., Ruck A., Martin K. F., Kendall D. A., Hill S. J. Histamine-induced changes in intracellular calcium in undifferentiated N1E-115 cells. Biochem Soc Trans. 1991 Feb;19(1):6S–6S. doi: 10.1042/bst019006s. [DOI] [PubMed] [Google Scholar]
  20. Kastritsis C. H., Salm A. K., McCarthy K. Stimulation of the P2Y purinergic receptor on type 1 astroglia results in inositol phosphate formation and calcium mobilization. J Neurochem. 1992 Apr;58(4):1277–1284. doi: 10.1111/j.1471-4159.1992.tb11339.x. [DOI] [PubMed] [Google Scholar]
  21. Koike M., Kashiwagura T., Takeguchi N. Gluconeogenesis stimulated by extracellular ATP is triggered by the initial increase in the intracellular Ca2+ concentration of the periphery of hepatocytes. Biochem J. 1992 Apr 1;283(Pt 1):265–272. doi: 10.1042/bj2830265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Monck J. R., Williamson R. E., Rogulja I., Fluharty S. J., Williamson J. R. Angiotensin II effects on the cytosolic free Ca2+ concentration in N1E-115 neuroblastoma cells: kinetic properties of the Ca2+ transient measured in single fura-2-loaded cells. J Neurochem. 1990 Jan;54(1):278–287. doi: 10.1111/j.1471-4159.1990.tb13312.x. [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. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  25. Pfeilschifter J. Comparison of extracellular ATP and UTP signalling in rat renal mesangial cells. No indications for the involvement of separate purino- and pyrimidino-ceptors. Biochem J. 1990 Dec 1;272(2):469–472. doi: 10.1042/bj2720469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Phaneuf S., Berta P., Casanova J., Cavadore J. C. ATP stimulates inositol phosphates accumulation and calcium mobilization in a primary culture of rat aortic myocytes. Biochem Biophys Res Commun. 1987 Mar 13;143(2):454–460. doi: 10.1016/0006-291x(87)91375-1. [DOI] [PubMed] [Google Scholar]
  27. Rice W. R., Singleton F. M. P2Y-purinoceptor regulation of surfactant secretion from rat isolated alveolar type II cells is associated with mobilization of intracellular calcium. Br J Pharmacol. 1987 Aug;91(4):833–838. doi: 10.1111/j.1476-5381.1987.tb11282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Richardson P. J., Brown S. J. ATP release from affinity-purified rat cholinergic nerve terminals. J Neurochem. 1987 Feb;48(2):622–630. doi: 10.1111/j.1471-4159.1987.tb04138.x. [DOI] [PubMed] [Google Scholar]
  29. Sato K., Okajima F., Kondo Y. Extracellular ATP stimulates three different receptor-signal transduction systems in FRTL-5 thyroid cells. Activation of phospholipase C, and inhibition and activation of adenylate cyclase. Biochem J. 1992 Apr 1;283(Pt 1):281–287. doi: 10.1042/bj2830281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stutchfield J., Cockcroft S. Undifferentiated HL60 cells respond to extracellular ATP and UTP by stimulating phospholipase C activation and exocytosis. FEBS Lett. 1990 Mar 26;262(2):256–258. doi: 10.1016/0014-5793(90)80204-v. [DOI] [PubMed] [Google Scholar]
  31. 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]

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

RESOURCES