Skip to main content
NCBI home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1991 Mar;434:647–660. doi: 10.1113/jphysiol.1991.sp018491

Comparison of adenosine triphosphate- and nicotine-activated inward currents in rat phaeochromocytoma cells.

K Nakazawa 1, K Fujimori 1, A Takanaka 1, K Inoue 1
PMCID: PMC1181439  PMID: 2023135

Abstract

1. The adenosine triphosphate (ATP)-activated inward current was compared to the nicotine-activated inward current in nerve growth factor (NGF)-treated rat phaeochromocytoma PC12 cells. 2. Both ATP and nicotine activated an inward current at negative holding potentials. The concentration of ATP necessary to activate the inward current was about 10-fold higher than that of nicotine; the EC50 was 20.5 microM for ATP and 2.4 microM for nicotine. The maximal responses induced by ATP and nicotine were almost identical in the same cells. The current-voltage relationship for the ATP-activated current was very similar to that for the nicotine-activated current, and both currents reversed around 0 mV in a physiological saline. 3. The ATP-activated current and the nicotine-activated current were not additive; the current activated by a combined administration of ATP (100 microM) and nicotine (10 microM) was only about 20% larger than the current activated by either ATP or nicotine alone. Nicotine (100 microM) did not increase the current activated by 1 microM-ATP. 4. ATP could activate an inward current in the cells even after desensitization to nicotine had developed. 5. Hexamethonium (100 microM) selectively blocked the nicotine-activated current whereas suramin (100 microM), a purinoceptor antagonist, selectively blocked the ATP-activated current. 6. Ionic selectivity was studied by changing compositions of extracellular solutions. When external Na+ was replaced with Cs+, both ATP and nicotine activated inward currents. However, with an extracellular solution containing Tris or glucosamine as a major cation, only ATP, not nicotine, activated an inward current. 7. ATP- and nicotine-activated currents were also recorded from cells bathed in a solution containing 1.8 mM-Ca2+ as the only external cation, suggesting that both pathways are Ca2+ permeable. 8. The results suggest that the ATP-sensitive ionic pathway is not independent of the nicotine-sensitive pathway in these cells. Our working hypothesis is that ATP and nicotine activate the same channels but the binding sites and the open-states of the channels are different between these two agonists.

Full text

PDF
647

Selected References

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

  1. Amy C. M., Bennett E. L. Increased sodium ion conductance through nicotinic acetylcholine receptor channels in PC12 cells exposed to nerve growth factors. J Neurosci. 1983 Aug;3(8):1547–1553. doi: 10.1523/JNEUROSCI.03-08-01547.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ascher P., Large W. A., Rang H. P. Studies on the mechanism of action of acetylcholine antagonists on rat parasympathetic ganglion cells. J Physiol. 1979 Oct;295:139–170. doi: 10.1113/jphysiol.1979.sp012958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bean B. P. ATP-activated channels in rat and bullfrog sensory neurons: concentration dependence and kinetics. J Neurosci. 1990 Jan;10(1):1–10. doi: 10.1523/JNEUROSCI.10-01-00001.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bean B. P., Friel D. D. ATP-activated channels in excitable cells. Ion Channels. 1990;2:169–203. doi: 10.1007/978-1-4615-7305-0_5. [DOI] [PubMed] [Google Scholar]
  5. Bean B. P., Williams C. A., Ceelen P. W. ATP-activated channels in rat and bullfrog sensory neurons: current-voltage relation and single-channel behavior. J Neurosci. 1990 Jan;10(1):11–19. doi: 10.1523/JNEUROSCI.10-01-00011.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benham C. D., Bolton T. B., Byrne N. G., Large W. A. Action of externally applied adenosine triphosphate on single smooth muscle cells dispersed from rabbit ear artery. J Physiol. 1987 Jun;387:473–488. doi: 10.1113/jphysiol.1987.sp016585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Brown D. A. M currents. Ion Channels. 1988;1:55–94. doi: 10.1007/978-1-4615-7302-9_2. [DOI] [PubMed] [Google Scholar]
  9. Burnstock G., Kennedy C. A dual function for adenosine 5'-triphosphate in the regulation of vascular tone. Excitatory cotransmitter with noradrenaline from perivascular nerves and locally released inhibitory intravascular agent. Circ Res. 1986 Mar;58(3):319–330. doi: 10.1161/01.res.58.3.319. [DOI] [PubMed] [Google Scholar]
  10. Colquhoun D., Hawkes A. G. On the stochastic properties of single ion channels. Proc R Soc Lond B Biol Sci. 1981 Mar 6;211(1183):205–235. doi: 10.1098/rspb.1981.0003. [DOI] [PubMed] [Google Scholar]
  11. Dichter M. A., Tischler A. S., Greene L. A. Nerve growth factor-induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature. 1977 Aug 11;268(5620):501–504. doi: 10.1038/268501a0. [DOI] [PubMed] [Google Scholar]
  12. Dunn P. M., Blakeley A. G. Suramin: a reversible P2-purinoceptor antagonist in the mouse vas deferens. Br J Pharmacol. 1988 Feb;93(2):243–245. doi: 10.1111/j.1476-5381.1988.tb11427.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friel D. D. An ATP-sensitive conductance in single smooth muscle cells from the rat vas deferens. J Physiol. 1988 Jul;401:361–380. doi: 10.1113/jphysiol.1988.sp017167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Honoré E., Martin C., Mironneau C., Mironneau J. An ATP-sensitive conductance in cultured smooth muscle cells from pregnant rat myometrium. Am J Physiol. 1989 Aug;257(2 Pt 1):C297–C305. doi: 10.1152/ajpcell.1989.257.2.C297. [DOI] [PubMed] [Google Scholar]
  17. Ifune C. K., Steinbach J. H. Regulation of sodium currents and acetylcholine responses in PC12 cells. Brain Res. 1990 Jan 8;506(2):243–248. doi: 10.1016/0006-8993(90)91257-h. [DOI] [PubMed] [Google Scholar]
  18. Igusa Y. Adenosine 5'-triphosphate activates acetylcholine receptor channels in cultured Xenopus myotomal muscle cells. J Physiol. 1988 Nov;405:169–185. doi: 10.1113/jphysiol.1988.sp017327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Kolb H. A., Wakelam M. J. Transmitter-like action of ATP on patched membranes of cultured myoblasts and myotubes. Nature. 1983 Jun 16;303(5918):621–623. doi: 10.1038/303621a0. [DOI] [PubMed] [Google Scholar]
  22. Krishtal O. A., Marchenko S. M., Obukhov A. G. Cationic channels activated by extracellular ATP in rat sensory neurons. Neuroscience. 1988 Dec;27(3):995–1000. doi: 10.1016/0306-4522(88)90203-5. [DOI] [PubMed] [Google Scholar]
  23. Krishtal O. A., Marchenko S. M., Obukhov A. G., Volkova T. M. Receptors for ATP in rat sensory neurones: the structure-function relationship for ligands. Br J Pharmacol. 1988 Dec;95(4):1057–1062. doi: 10.1111/j.1476-5381.1988.tb11739.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kurachi Y., Nakajima T., Sugimoto T. On the mechanism of activation of muscarinic K+ channels by adenosine in isolated atrial cells: involvement of GTP-binding proteins. Pflugers Arch. 1986 Sep;407(3):264–274. doi: 10.1007/BF00585301. [DOI] [PubMed] [Google Scholar]
  25. Lerma J., Kushner L., Zukin R. S., Bennett M. V. N-methyl-D-aspartate activates different channels than do kainate and quisqualate. Proc Natl Acad Sci U S A. 1989 Mar;86(6):2083–2087. doi: 10.1073/pnas.86.6.2083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mitsuka M., Hatanaka H. Increase of carbamylcholine-induced 22Na+ influx into pheochromocytoma PC12h cells by nerve growth factor. Brain Res. 1984 Feb;314(2):255–260. doi: 10.1016/0165-3806(84)90047-6. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Nakazawa K., Fujimori K., Takanaka A., Inoue K. Existence of muscarinic suppression of a K current in PC-12 pheochromocytoma cells. Am J Physiol. 1989 Nov;257(5 Pt 1):C1030–C1033. doi: 10.1152/ajpcell.1989.257.5.C1030. [DOI] [PubMed] [Google Scholar]
  29. Nakazawa K., Fujimori K., Takanaka A., Inoue K. Reversible and selective antagonism by suramin of ATP-activated inward current in PC12 phaeochromocytoma cells. Br J Pharmacol. 1990 Sep;101(1):224–226. doi: 10.1111/j.1476-5381.1990.tb12117.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nakazawa K., Matsuki N. Adenosine triphosphate-activated inward current in isolated smooth muscle cells from rat vas deferens. Pflugers Arch. 1987 Aug;409(6):644–646. doi: 10.1007/BF00584668. [DOI] [PubMed] [Google Scholar]
  31. O'Lague P. H., Huttner S. L., Vandenberg C. A., Morrison-Graham K., Horn R. Morphological properties and membrane channels of the growth cones induced in PC12 cells by nerve growth factor. J Neurosci Res. 1985;13(1-2):301–321. doi: 10.1002/jnr.490130120. [DOI] [PubMed] [Google Scholar]
  32. Rang H. P., Colquhoun D., Rang H. P. The action of ganglionic blocking drugs on the synaptic responses of rat submandibular ganglion cells. Br J Pharmacol. 1982 Jan;75(1):151–168. doi: 10.1111/j.1476-5381.1982.tb08768.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Streit J., Lux H. D. Voltage dependent calcium currents in PC12 growth cones and cells during NGF-induced cell growth. Pflugers Arch. 1987 May;408(6):634–641. doi: 10.1007/BF00581167. [DOI] [PubMed] [Google Scholar]
  34. den Hertog A., Nelemans A., Van den Akker J. The inhibitory action of suramin on the P2-purinoceptor response in smooth muscle cells of guinea-pig taenia caeci. Eur J Pharmacol. 1989 Aug 3;166(3):531–534. doi: 10.1016/0014-2999(89)90370-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES