Skip to main content
PMC home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1993 Oct;470:55–72. doi: 10.1113/jphysiol.1993.sp019847

ATP modulation of calcium channels in chromaffin cells.

L Gandía 1, A G García 1, M Morad 1
PMCID: PMC1143906  PMID: 8308743

Abstract

1. The effects of externally applied micromolar concentrations of adenosine 5'-triphosphate (ATP) on Ca2+ currents (ICa) were studied in whole-cell clamped adrenaline-secreting chromaffin cells. 2. Ca2+ currents in chromaffin cells activated at about -40 mV, reached a maximum at 0 mV and had an apparent reversal potential at +50 to +60 mV, indicating the existence of only high voltage-activated Ca2+ channels. 3. ATP blocked Ca2+ current rapidly, reversibly and in a concentration-dependent manner (10(-9)-10(-4) M). 4. ATP did not completely block Ca2+ current even at the highest concentrations used (100 microM). The remaining component of Ca2+ current was characterized by slower activation and inactivation kinetics. 5. ATP blocked ICa even in the presence of nisoldipine and/or omega-conotoxin GVIA, suggesting that its modulatory role is not specific for L- and/or N-type Ca2+ channels. 6. Other adenine nucleotides also blocked the Ca2+ current partially. The order of potencies was ATP > or = ADP > AMP >> adenosine, indicating that the ATP effects are most probably mediated by a P2-type purinergic receptor. 7. Dialysis of the cells with an intracellular solution containing 1 mM guanosine 5'-O-thiodiphosphate (GDP-beta-S) or pre-incubation of the cells with pertussis toxin (PTX) blocked the inhibitory effects of ATP. 8. Intracellular application of the non-hydrolysable GTP analogue guanosine 5'-O-(3'-thiotriphosphate) (GTP-gamma-S; 50 microM) also decreased ICa in a manner similar to that seen for ATP and significantly reduced the ATP inhibitory effect. 9. Conditioning pulses to potentials positive to +80 mV partly reversed the inhibitory effects of ATP on the Ca2+ current. The prepulse-induced enhancement of ICa depended on [GTP]i-related G protein activity such that concentrations larger than 200 microM GTP, or GTP-gamma-S (50 microM) were required for significant prepulse potentiation of the Ca2+ current, while dialysis with GDP-beta-S prevented it. 10. We conclude that the ATP, co-released with catecholamines in the intact adrenal gland, may inhibit the secretory process by down-regulating the Ca2+ channel via a P2-type purinergic receptor coupled to a PTX-sensitive G protein.

Full text

PDF
56

Selected References

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

  1. Aicardi G., Pollo A., Sher E., Carbone E. Noradrenergic inhibition and voltage-dependent facilitation of omega-conotoxin-sensitive Ca channels in insulin-secreting RINm5F cells. FEBS Lett. 1991 Apr 9;281(1-2):201–204. doi: 10.1016/0014-5793(91)80393-h. [DOI] [PubMed] [Google Scholar]
  2. Artalejo C. R., Dahmer M. K., Perlman R. L., Fox A. P. Two types of Ca2+ currents are found in bovine chromaffin cells: facilitation is due to the recruitment of one type. J Physiol. 1991 Jan;432:681–707. doi: 10.1113/jphysiol.1991.sp018406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Artalejo C. R., Mogul D. J., Perlman R. L., Fox A. P. Three types of bovine chromaffin cell Ca2+ channels: facilitation increases the opening probability of a 27 pS channel. J Physiol. 1991 Dec;444:213–240. doi: 10.1113/jphysiol.1991.sp018874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Artalejo C. R., Perlman R. L., Fox A. P. Omega-conotoxin GVIA blocks a Ca2+ current in bovine chromaffin cells that is not of the "classic" N type. Neuron. 1992 Jan;8(1):85–95. doi: 10.1016/0896-6273(92)90110-y. [DOI] [PubMed] [Google Scholar]
  5. Artalejo C. R., Rossie S., Perlman R. L., Fox A. P. Voltage-dependent phosphorylation may recruit Ca2+ current facilitation in chromaffin cells. Nature. 1992 Jul 2;358(6381):63–66. doi: 10.1038/358063a0. [DOI] [PubMed] [Google Scholar]
  6. Bean B. P. Neurotransmitter inhibition of neuronal calcium currents by changes in channel voltage dependence. Nature. 1989 Jul 13;340(6229):153–156. doi: 10.1038/340153a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Bossu J. L., De Waard M., Feltz A. Inactivation characteristics reveal two calcium currents in adult bovine chromaffin cells. J Physiol. 1991 Jun;437:603–620. doi: 10.1113/jphysiol.1991.sp018614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burnstock G. Overview. Purinergic mechanisms. Ann N Y Acad Sci. 1990;603:1–18. doi: 10.1111/j.1749-6632.1990.tb37657.x. [DOI] [PubMed] [Google Scholar]
  10. Callewaert G., Johnson R. G., Morad M. Regulation of the secretory response in bovine chromaffin cells. Am J Physiol. 1991 Apr;260(4 Pt 1):C851–C860. doi: 10.1152/ajpcell.1991.260.4.C851. [DOI] [PubMed] [Google Scholar]
  11. Castillo C. J., Moro M. A., Del Valle M., Sillero A., García A. G., Sillero M. A. Diadenosine tetraphosphate is co-released with ATP and catecholamines from bovine adrenal medulla. J Neurochem. 1992 Aug;59(2):723–732. doi: 10.1111/j.1471-4159.1992.tb09428.x. [DOI] [PubMed] [Google Scholar]
  12. Ceña V., Brocklehurst K. W., Pollard H. B., Rojas E. Pertussis toxin stimulation of catecholamine release from adrenal medullary chromaffin cells: mechanism may be by direct activation of L-type and G-type calcium channels. J Membr Biol. 1991 May;122(1):23–31. doi: 10.1007/BF01872736. [DOI] [PubMed] [Google Scholar]
  13. Chern Y. J., Herrera M., Kao L. S., Westhead E. W. Inhibition of catecholamine secretion from bovine chromaffin cells by adenine nucleotides and adenosine. J Neurochem. 1987 May;48(5):1573–1576. doi: 10.1111/j.1471-4159.1987.tb05703.x. [DOI] [PubMed] [Google Scholar]
  14. Chern Y. J., Kim K. T., Slakey L. L., Westhead E. W. Adenosine receptors activate adenylate cyclase and enhance secretion from bovine adrenal chromaffin cells in the presence of forskolin. J Neurochem. 1988 May;50(5):1484–1493. doi: 10.1111/j.1471-4159.1988.tb03034.x. [DOI] [PubMed] [Google Scholar]
  15. Diverse-Pierluissi M., Dunlap K., Westhead E. W. Multiple actions of extracellular ATP on calcium currents in cultured bovine chromaffin cells. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1261–1265. doi: 10.1073/pnas.88.4.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Douglas W. W., Poisner A. M., Rubin R. P. Efflux of adenine nucleotides from perfused adrenal glands exposed to nicotine and other chromaffin cell stimulants. J Physiol. 1965 Jul;179(1):130–137. doi: 10.1113/jphysiol.1965.sp007652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dubyak G. R. Signal transduction by P2-purinergic receptors for extracellular ATP. Am J Respir Cell Mol Biol. 1991 Apr;4(4):295–300. doi: 10.1165/ajrcmb/4.4.295. [DOI] [PubMed] [Google Scholar]
  18. Eckert R., Chad J. E. Inactivation of Ca channels. Prog Biophys Mol Biol. 1984;44(3):215–267. doi: 10.1016/0079-6107(84)90009-9. [DOI] [PubMed] [Google Scholar]
  19. Fenwick E. M., Marty A., Neher E. Sodium and calcium channels in bovine chromaffin cells. J Physiol. 1982 Oct;331:599–635. doi: 10.1113/jphysiol.1982.sp014394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gandía L., Casado L. F., López M. G., García A. G. Separation of two pathways for calcium entry into chromaffin cells. Br J Pharmacol. 1991 May;103(1):1073–1078. doi: 10.1111/j.1476-5381.1991.tb12302.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grassi F., Lux H. D. Voltage-dependent GABA-induced modulation of calcium currents in chick sensory neurons. Neurosci Lett. 1989 Oct 23;105(1-2):113–119. doi: 10.1016/0304-3940(89)90021-9. [DOI] [PubMed] [Google Scholar]
  22. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  23. Hans M., Illes P., Takeda K. The blocking effects of omega-conotoxin on Ca current in bovine chromaffin cells. Neurosci Lett. 1990 Jun 22;114(1):63–68. doi: 10.1016/0304-3940(90)90429-d. [DOI] [PubMed] [Google Scholar]
  24. Hescheler J., Rosenthal W., Trautwein W., Schultz G. The GTP-binding protein, Go, regulates neuronal calcium channels. 1987 Jan 29-Feb 4Nature. 325(6103):445–447. doi: 10.1038/325445a0. [DOI] [PubMed] [Google Scholar]
  25. Holz G. G., 4th, Rane S. G., Dunlap K. GTP-binding proteins mediate transmitter inhibition of voltage-dependent calcium channels. Nature. 1986 Feb 20;319(6055):670–672. doi: 10.1038/319670a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hoshi T., Rothlein J., Smith S. J. Facilitation of Ca2+-channel currents in bovine adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5871–5875. doi: 10.1073/pnas.81.18.5871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hoshi T., Smith S. J. Large depolarization induces long openings of voltage-dependent calcium channels in adrenal chromaffin cells. J Neurosci. 1987 Feb;7(2):571–580. doi: 10.1523/JNEUROSCI.07-02-00571.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Johnson R. G., Jr Proton pumps and chemiosmotic coupling as a generalized mechanism for neurotransmitter and hormone transport. Ann N Y Acad Sci. 1987;493:162–177. doi: 10.1111/j.1749-6632.1987.tb27198.x. [DOI] [PubMed] [Google Scholar]
  29. Kasai H., Aosaki T. Modulation of Ca-channel current by an adenosine analog mediated by a GTP-binding protein in chick sensory neurons. Pflugers Arch. 1989 Jun;414(2):145–149. doi: 10.1007/BF00580956. [DOI] [PubMed] [Google Scholar]
  30. Kim K. T., Westhead E. W. Cellular responses to Ca2+ from extracellular and intracellular sources are different as shown by simultaneous measurements of cytosolic Ca2+ and secretion from bovine chromaffin cells. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9881–9885. doi: 10.1073/pnas.86.24.9881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kleuss C., Hescheler J., Ewel C., Rosenthal W., Schultz G., Wittig B. Assignment of G-protein subtypes to specific receptors inducing inhibition of calcium currents. Nature. 1991 Sep 5;353(6339):43–48. doi: 10.1038/353043a0. [DOI] [PubMed] [Google Scholar]
  32. Lipscombe D., Kongsamut S., Tsien R. W. Alpha-adrenergic inhibition of sympathetic neurotransmitter release mediated by modulation of N-type calcium-channel gating. Nature. 1989 Aug 24;340(6235):639–642. doi: 10.1038/340639a0. [DOI] [PubMed] [Google Scholar]
  33. Lopez H. S., Brown A. M. Correlation between G protein activation and reblocking kinetics of Ca2+ channel currents in rat sensory neurons. Neuron. 1991 Dec;7(6):1061–1068. doi: 10.1016/0896-6273(91)90350-9. [DOI] [PubMed] [Google Scholar]
  34. Moro M. A., López M. G., Gandía L., Michelena P., García A. G. Separation and culture of living adrenaline- and noradrenaline-containing cells from bovine adrenal medullae. Anal Biochem. 1990 Mar;185(2):243–248. doi: 10.1016/0003-2697(90)90287-j. [DOI] [PubMed] [Google Scholar]
  35. Nakazawa K., Fujimori K., Takanaka A., Inoue K. An ATP-activated conductance in pheochromocytoma cells and its suppression by extracellular calcium. J Physiol. 1990 Sep;428:257–272. doi: 10.1113/jphysiol.1990.sp018211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nakazawa K., Inoue K., Fujimori K., Takanaka A. ATP-activated single-channel currents recorded from cell-free patches of pheochromocytoma PC12 cells. Neurosci Lett. 1990 Oct 30;119(1):5–8. doi: 10.1016/0304-3940(90)90741-q. [DOI] [PubMed] [Google Scholar]
  37. Pintor J., Torres M., Castro E., Miras-Portugal M. T. Characterization of diadenosine tetraphosphate (Ap4A) binding sites in cultured chromaffin cells: evidence for a P2y site. Br J Pharmacol. 1991 Aug;103(4):1980–1984. doi: 10.1111/j.1476-5381.1991.tb12363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rubin R. P., Jaanus S. D. The release of nucleotide from the adrenal medulla by indirectly acting sympathomimetic amines. Biochem Pharmacol. 1967 Jun;16(6):1007–1012. doi: 10.1016/0006-2952(67)90273-0. [DOI] [PubMed] [Google Scholar]
  39. Scott R. H., Dolphin A. C. Regulation of calcium currents by a GTP analogue: potentiation of (-)-baclofen-mediated inhibition. Neurosci Lett. 1986 Aug 15;69(1):59–64. doi: 10.1016/0304-3940(86)90414-3. [DOI] [PubMed] [Google Scholar]
  40. Sorbera L. A., Morad M. Modulation of cardiac sodium channels by cAMP receptors on the myocyte surface. Science. 1991 Sep 13;253(5025):1286–1289. doi: 10.1126/science.1653970. [DOI] [PubMed] [Google Scholar]
  41. Stevens P., Robinson R. L., Van Dyke K., Stitxel R. Synthesis, storage and drug-induced release of atp-8-3h in the perfused bovine adrenal gland. Pharmacology. 1975;13(1):40–55. doi: 10.1159/000136883. [DOI] [PubMed] [Google Scholar]
  42. Swandulla D., Carbone E., Lux H. D. Do calcium channel classifications account for neuronal calcium channel diversity? Trends Neurosci. 1991 Feb;14(2):46–51. doi: 10.1016/0166-2236(91)90018-p. [DOI] [PubMed] [Google Scholar]
  43. Winkler H., Westhead E. The molecular organization of adrenal chromaffin granules. Neuroscience. 1980;5(11):1803–1823. doi: 10.1016/0306-4522(80)90031-7. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES