Skip to main content
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1995 Aug;115(8):1502–1508. doi: 10.1111/j.1476-5381.1995.tb16643.x

Inhibition by Zn2+ of uridine 5'-triphosphate-induced Ca(2+)-influx but not Ca(2+)-mobilization in rat phaeochromocytoma cells.

S Koizumi 1, K Nakazawa 1, K Inoue 1
PMCID: PMC1908867  PMID: 8564211

Abstract

1. Uridine 5'-triphosphate (UTP)-evoked increase in intracellular Ca2+ concentration ([Ca]i) and release of dopamine were investigated in rat phaeochromocytoma PC12 cells. UTP (1-100 microM) evoked an increase in [Ca]i in a concentration-dependent manner. This response was decreased to about 30% by extracellular Ca(2+)-depletion, but not abolished. This [Ca]i rise was mimicked by 100 microM ATP but not by 100 microM 2-methyl-thio-ATP or alpha,beta-methylene-ATP in the absence of external Ca2+, suggesting that the response was mediated by P2U purinoceptors, a subclass of P2-purinoceptors. 2. The UTP-evoked [Ca]i rise consisted of two components; a transient and a sustained one. When external Ca2+ was removed, the sustained component was abolished while the transient component was decreased by about 70% but did not disappear. These results suggest that UTP induces Ca(2+)-mobilization and, subsequently, Ca(2+)-influx. 3. The UTP-evoked increase in [Ca]i was not affected by Cd2+ (100 and 300 microM) or nicardipine (30 microM), inhibitors of voltage-gated calcium channels, but was significantly inhibited by Zn2+ (10-300 microM) in the presence of external Ca2+. Zn2+, however, did not affect the Ca2+ response to UTP in the absence of external Ca2+. 4. UTP (30 microM-1 mM) evoked the release of dopamine from the cells in a concentration-dependent manner. This dopamine release was abolished by Ca(2+)-depletion or Zn2+ but not by Cd2+ or nicardipine. 5. Taken together, the data demonstrate that UTP stimulates P2U-purinoceptors and induces a rise in [Ca]i both by Ca(2+)-mobilization and Ca(2+)-influx in PC12 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Full text

PDF
1504

Selected References

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

  1. Abbracchio M. P., Burnstock G. Purinoceptors: are there families of P2X and P2Y purinoceptors? Pharmacol Ther. 1994;64(3):445–475. doi: 10.1016/0163-7258(94)00048-4. [DOI] [PubMed] [Google Scholar]
  2. Barry V. A., Cheek T. R. Extracellular ATP triggers two functionally distinct calcium signalling pathways in PC12 cells. J Cell Sci. 1994 Feb;107(Pt 2):451–462. doi: 10.1242/jcs.107.2.451. [DOI] [PubMed] [Google Scholar]
  3. Bean B. P. Pharmacology and electrophysiology of ATP-activated ion channels. Trends Pharmacol Sci. 1992 Mar;13(3):87–90. doi: 10.1016/0165-6147(92)90032-2. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Clementi E., Scheer H., Zacchetti D., Fasolato C., Pozzan T., Meldolesi J. Receptor-activated Ca2+ influx. Two independently regulated mechanisms of influx stimulation coexist in neurosecretory PC12 cells. J Biol Chem. 1992 Feb 5;267(4):2164–2172. [PubMed] [Google Scholar]
  7. Cloues R., Jones S., Brown D. A. Zn2+ potentiates ATP-activated currents in rat sympathetic neurons. Pflugers Arch. 1993 Jul;424(2):152–158. doi: 10.1007/BF00374606. [DOI] [PubMed] [Google Scholar]
  8. Edwards F. A., Gibb A. J., Colquhoun D. ATP receptor-mediated synaptic currents in the central nervous system. Nature. 1992 Sep 10;359(6391):144–147. doi: 10.1038/359144a0. [DOI] [PubMed] [Google Scholar]
  9. Evans R. J., Derkach V., Surprenant A. ATP mediates fast synaptic transmission in mammalian neurons. Nature. 1992 Jun 11;357(6378):503–505. doi: 10.1038/357503a0. [DOI] [PubMed] [Google Scholar]
  10. Frederickson C. J. Neurobiology of zinc and zinc-containing neurons. Int Rev Neurobiol. 1989;31:145–238. doi: 10.1016/s0074-7742(08)60279-2. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Greene L. A., Tischler A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. doi: 10.1073/pnas.73.7.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Harrison N. L., Gibbons S. J. Zn2+: an endogenous modulator of ligand- and voltage-gated ion channels. Neuropharmacology. 1994 Aug;33(8):935–952. doi: 10.1016/0028-3908(94)90152-x. [DOI] [PubMed] [Google Scholar]
  15. Inoue K., Kenimer J. G. Muscarinic stimulation of calcium influx and norepinephrine release in PC12 cells. J Biol Chem. 1988 Jun 15;263(17):8157–8161. [PubMed] [Google Scholar]
  16. Inoue K., Koizumi S., Nakazawa K. Glutamate-evoked release of adenosine 5'-triphosphate causing an increase in intracellular calcium in hippocampal neurons. Neuroreport. 1995 Feb 15;6(3):437–440. doi: 10.1097/00001756-199502000-00008. [DOI] [PubMed] [Google Scholar]
  17. Inoue K., Nakazawa K., Fujimori K., Takanaka A. Extracellular adenosine 5'-triphosphate-evoked norepinephrine secretion not relating to voltage-gated Ca channels in pheochromocytoma PC12 cells. Neurosci Lett. 1989 Dec 4;106(3):294–299. doi: 10.1016/0304-3940(89)90179-1. [DOI] [PubMed] [Google Scholar]
  18. Inoue K., Nakazawa K., Fujimori K., Watano T., Takanaka A. Extracellular adenosine 5'-triphosphate-evoked glutamate release in cultured hippocampal neurons. Neurosci Lett. 1992 Jan 6;134(2):215–218. doi: 10.1016/0304-3940(92)90520-h. [DOI] [PubMed] [Google Scholar]
  19. Koizumi S., Ikeda M., Inoue K., Nakazawa K., Inoue K. Enhancement by zinc of ATP-evoked dopamine release from rat pheochromocytoma PC12 cells. Brain Res. 1995 Feb 27;673(1):75–82. doi: 10.1016/0006-8993(94)01404-6. [DOI] [PubMed] [Google Scholar]
  20. Koizumi S., Watano T., Nakazawa K., Inoue K. Potentiation by adenosine of ATP-evoked dopamine release via a pertussis toxin-sensitive mechanism in rat phaeochromocytoma PC12 cells. Br J Pharmacol. 1994 Jul;112(3):992–997. doi: 10.1111/j.1476-5381.1994.tb13179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Li C., Peoples R. W., Li Z., Weight F. F. Zn2+ potentiates excitatory action of ATP on mammalian neurons. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8264–8267. doi: 10.1073/pnas.90.17.8264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin T. A., Lustig K. D., Sportiello M. G., Weisman G. A., Sun G. Y. Signal transduction pathways coupled to a P2U receptor in neuroblastoma x glioma (NG108-15) cells. J Neurochem. 1993 Mar;60(3):1115–1125. doi: 10.1111/j.1471-4159.1993.tb03262.x. [DOI] [PubMed] [Google Scholar]
  23. Lustig K. D., Shiau A. K., Brake A. J., Julius D. Expression cloning of an ATP receptor from mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5113–5117. doi: 10.1073/pnas.90.11.5113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Majid M. A., Okajima F., Kondo Y. UTP activates phospholipase C-Ca2+ system through a receptor different from the 53-kDa ATP receptor in PC12 cells. Biochem Biophys Res Commun. 1993 Aug 31;195(1):415–421. doi: 10.1006/bbrc.1993.2059. [DOI] [PubMed] [Google Scholar]
  25. Munshi R., DeBernardi M. A., Brooker G. P2U-purinergic receptors on C6-2B rat glioma cells: modulation of cytosolic Ca2+ and cAMP levels by protein kinase C. Mol Pharmacol. 1993 Dec;44(6):1185–1191. [PubMed] [Google Scholar]
  26. 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]
  27. Nakazawa K., Inoue K. Roles of Ca2+ influx through ATP-activated channels in catecholamine release from pheochromocytoma PC12 cells. J Neurophysiol. 1992 Dec;68(6):2026–2032. doi: 10.1152/jn.1992.68.6.2026. [DOI] [PubMed] [Google Scholar]
  28. Nikodijevic B., Sei Y., Shin Y., Daly J. W. Effects of ATP and UTP in pheochromocytoma PC12 cells: evidence for the presence of three P2 receptors, only one of which subserves stimulation of norepinephrine release. Cell Mol Neurobiol. 1994 Feb;14(1):27–47. doi: 10.1007/BF02088587. [DOI] [PubMed] [Google Scholar]
  29. Ohara-Imaizumi M., Nakazawa K., Obama T., Fujimori K., Takanaka A., Inoue K. Inhibitory action of peripheral-type benzodiazepines on dopamine release from PC12 pheochromocytoma cells. J Pharmacol Exp Ther. 1991 Nov;259(2):484–489. [PubMed] [Google Scholar]
  30. 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]
  31. Putney J. W., Jr A model for receptor-regulated calcium entry. Cell Calcium. 1986 Feb;7(1):1–12. doi: 10.1016/0143-4160(86)90026-6. [DOI] [PubMed] [Google Scholar]
  32. Raha S., de Souza L. R., Reed J. K. Intracellular signalling by nucleotide receptors in PC12 pheochromocytoma cells. J Cell Physiol. 1993 Mar;154(3):623–630. doi: 10.1002/jcp.1041540322. [DOI] [PubMed] [Google Scholar]
  33. Randriamampita C., Tsien R. Y. Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature. 1993 Aug 26;364(6440):809–814. doi: 10.1038/364809a0. [DOI] [PubMed] [Google Scholar]
  34. Rassendren F. A., Lory P., Pin J. P., Nargeot J. Zinc has opposite effects on NMDA and non-NMDA receptors expressed in Xenopus oocytes. Neuron. 1990 May;4(5):733–740. doi: 10.1016/0896-6273(90)90199-p. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. 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]

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

RESOURCES