Skip to main content
PMC home page
The Journal of General Physiology logoLink to The Journal of General Physiology
. 1995 Jun 1;105(6):701–723. doi: 10.1085/jgp.105.6.701

Monovalent and divalent cation permeability and block of neuronal nicotinic receptor channels in rat parasympathetic ganglia

PMCID: PMC2216957  PMID: 7561740

Abstract

Acetylcholine-evoked currents mediated by activation of nicotinic receptors in rat parasympathetic neurons were examined using whole-cell voltage clamp. The relative permeability of the neuronal nicotinic acetylcholine (nACh) receptor channel to monovalent and divalent inorganic and organic cations was determined from reversal potential measurements. The channel exhibited weak selectivity among the alkali metals with a selectivity sequence of Cs+ > K+ > Rb+ > Na+ > Li+, and permeability ratios relative to Na+ (Px/PNa) ranging from 1.27 to 0.75. The selectivity of the alkaline earths was also weak, with the sequence of Mg2+ > Sr2+ > Ba2+ > Ca2+, and relative permeabilities of 1.10 to 0.65. The relative Ca2+ permeability (PCa/PNa) of the neuronal nACh receptor channel is approximately fivefold higher than that of the motor endplate channel (Adams, D. J., T. M. Dwyer, and B. Hille. 1980. Journal of General Physiology. 75:493-510). The transition metal cation, Mn2+ was permeant (Px/PNa = 0.67), whereas Ni2+, Zn2+, and Cd2+ blocked ACh-evoked currents with half-maximal inhibition (IC50) occurring at approximately 500 microM, 5 microM and 1 mM, respectively. In contrast to the muscle endplate AChR channel, that at least 56 organic cations which are permeable to (Dwyer et al., 1980), the majority of organic cations tested were found to completely inhibit ACh- evoked currents in rat parasympathetic neurons. Concentration-response curves for guanidinium, ethylammonium, diethanolammonium and arginine inhibition of ACh-evoked currents yielded IC50's of approximately 2.5- 6.0 mM. The organic cations, hydrazinium, methylammonium, ethanolammonium and Tris, were measureably permeant, and permeability ratios varied inversely with the molecular size of the cation. Modeling suggests that the pore has a minimum diameter of 7.6 A. Thus, there are substantial differences in ion permeation and block between the nACh receptor channels of mammalian parasympathetic neurons and amphibian skeletal muscle which represent functional consequences of differences in the primary structure of the subunits of the ACh receptor channel.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Adams D. J., Dwyer T. M., Hille B. The permeability of endplate channels to monovalent and divalent metal cations. J Gen Physiol. 1980 May;75(5):493–510. doi: 10.1085/jgp.75.5.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adams D. J., Nonner W., Dwyer T. M., Hille B. Block of endplate channels by permeant cations in frog skeletal muscle. J Gen Physiol. 1981 Dec;78(6):593–615. doi: 10.1085/jgp.78.6.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Adams D. J., Nutter T. J. Calcium permeability and modulation of nicotinic acetylcholine receptor-channels in rat parasympathetic neurons. J Physiol Paris. 1992;86(1-3):67–76. doi: 10.1016/s0928-4257(05)80009-9. [DOI] [PubMed] [Google Scholar]
  4. Aibara K., Akaike N. Acetylcholine-activated ionic currents in isolated paratracheal ganglion cells of the rat. Brain Res. 1991 Aug 30;558(1):20–26. doi: 10.1016/0006-8993(91)90709-5. [DOI] [PubMed] [Google Scholar]
  5. Anand R., Conroy W. G., Schoepfer R., Whiting P., Lindstrom J. Neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes have a pentameric quaternary structure. J Biol Chem. 1991 Jun 15;266(17):11192–11198. [PubMed] [Google Scholar]
  6. Butler J. N. The thermodynamic activity of calcium ion in sodium chloride-calcium chloride electrolytes. Biophys J. 1968 Dec;8(12):1426–1433. doi: 10.1016/S0006-3495(68)86564-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen B. N., Labarca C., Czyzyk L., Davidson N., Lester H. A. Tris+/Na+ permeability ratios of nicotinic acetylcholine receptors are reduced by mutations near the intracellular end of the M2 region. J Gen Physiol. 1992 Apr;99(4):545–572. doi: 10.1085/jgp.99.4.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen B. N., Labarca C., Davidson N., Lester H. A. Mutations in M2 alter the selectivity of the mouse nicotinic acetylcholine receptor for organic and alkali metal cations. J Gen Physiol. 1992 Sep;100(3):373–400. doi: 10.1085/jgp.100.3.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cooper E., Couturier S., Ballivet M. Pentameric structure and subunit stoichiometry of a neuronal nicotinic acetylcholine receptor. Nature. 1991 Mar 21;350(6315):235–238. doi: 10.1038/350235a0. [DOI] [PubMed] [Google Scholar]
  10. Dani J. A., Eisenman G. Monovalent and divalent cation permeation in acetylcholine receptor channels. Ion transport related to structure. J Gen Physiol. 1987 Jun;89(6):959–983. doi: 10.1085/jgp.89.6.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Decker E. R., Dani J. A. Calcium permeability of the nicotinic acetylcholine receptor: the single-channel calcium influx is significant. J Neurosci. 1990 Oct;10(10):3413–3420. doi: 10.1523/JNEUROSCI.10-10-03413.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dwyer T. M., Adams D. J., Hille B. The permeability of the endplate channel to organic cations in frog muscle. J Gen Physiol. 1980 May;75(5):469–492. doi: 10.1085/jgp.75.5.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. EISENMAN G. Cation selective glass electrodes and their mode of operation. Biophys J. 1962 Mar;2(2 Pt 2):259–323. doi: 10.1016/s0006-3495(62)86959-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fieber L. A., Adams D. J. Acetylcholine-evoked currents in cultured neurones dissociated from rat parasympathetic cardiac ganglia. J Physiol. 1991 Mar;434:215–237. doi: 10.1113/jphysiol.1991.sp018466. [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. Hille B. Ionic selectivity of Na and K channels of nerve membranes. Membranes. 1975;3:255–323. [PubMed] [Google Scholar]
  17. Imoto K., Busch C., Sakmann B., Mishina M., Konno T., Nakai J., Bujo H., Mori Y., Fukuda K., Numa S. Rings of negatively charged amino acids determine the acetylcholine receptor channel conductance. Nature. 1988 Oct 13;335(6191):645–648. doi: 10.1038/335645a0. [DOI] [PubMed] [Google Scholar]
  18. Imoto K., Methfessel C., Sakmann B., Mishina M., Mori Y., Konno T., Fukuda K., Kurasaki M., Bujo H., Fujita Y. Location of a delta-subunit region determining ion transport through the acetylcholine receptor channel. Nature. 1986 Dec 18;324(6098):670–674. doi: 10.1038/324670a0. [DOI] [PubMed] [Google Scholar]
  19. Konno T., Busch C., Von Kitzing E., Imoto K., Wang F., Nakai J., Mishina M., Numa S., Sakmann B. Rings of anionic amino acids as structural determinants of ion selectivity in the acetylcholine receptor channel. Proc Biol Sci. 1991 May 22;244(1310):69–79. doi: 10.1098/rspb.1991.0053. [DOI] [PubMed] [Google Scholar]
  20. Landau E. M., Gavish B., Nachshen D. A., Lotan I. pH dependence of the acetylcholine receptor channel: a species variation. J Gen Physiol. 1981 Jun;77(6):647–666. doi: 10.1085/jgp.77.6.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lewis C. A. Divalent cation effects on acetylcholine-activated channels at the frog neuromuscular junction. Cell Mol Neurobiol. 1984 Sep;4(3):273–284. doi: 10.1007/BF00733590. [DOI] [PubMed] [Google Scholar]
  22. Lewis C. A. Ion-concentration dependence of the reversal potential and the single channel conductance of ion channels at the frog neuromuscular junction. J Physiol. 1979 Jan;286:417–445. doi: 10.1113/jphysiol.1979.sp012629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lipton S. A., Aizenman E., Loring R. H. Neural nicotinic acetylcholine responses in solitary mammalian retinal ganglion cells. Pflugers Arch. 1987 Sep;410(1-2):37–43. doi: 10.1007/BF00581893. [DOI] [PubMed] [Google Scholar]
  24. Luetje C. W., Patrick J. Both alpha- and beta-subunits contribute to the agonist sensitivity of neuronal nicotinic acetylcholine receptors. J Neurosci. 1991 Mar;11(3):837–845. doi: 10.1523/JNEUROSCI.11-03-00837.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Luetje C. W., Wada K., Rogers S., Abramson S. N., Tsuji K., Heinemann S., Patrick J. Neurotoxins distinguish between different neuronal nicotinic acetylcholine receptor subunit combinations. J Neurochem. 1990 Aug;55(2):632–640. doi: 10.1111/j.1471-4159.1990.tb04180.x. [DOI] [PubMed] [Google Scholar]
  26. Lukas R. J., Bencherif M. Heterogeneity and regulation of nicotinic acetylcholine receptors. Int Rev Neurobiol. 1992;34:25–131. doi: 10.1016/s0074-7742(08)60097-5. [DOI] [PubMed] [Google Scholar]
  27. Mathie A., Cull-Candy S. G., Colquhoun D. Conductance and kinetic properties of single nicotinic acetylcholine receptor channels in rat sympathetic neurones. J Physiol. 1991 Aug;439:717–750. doi: 10.1113/jphysiol.1991.sp018690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mayer M. L., Westbrook G. L. Permeation and block of N-methyl-D-aspartic acid receptor channels by divalent cations in mouse cultured central neurones. J Physiol. 1987 Dec;394:501–527. doi: 10.1113/jphysiol.1987.sp016883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mayer M. L., Westbrook G. L. The action of N-methyl-D-aspartic acid on mouse spinal neurones in culture. J Physiol. 1985 Apr;361:65–90. doi: 10.1113/jphysiol.1985.sp015633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mulle C., Changeux J. P. A novel type of nicotinic receptor in the rat central nervous system characterized by patch-clamp techniques. J Neurosci. 1990 Jan;10(1):169–175. doi: 10.1523/JNEUROSCI.10-01-00169.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mulle C., Choquet D., Korn H., Changeux J. P. Calcium influx through nicotinic receptor in rat central neurons: its relevance to cellular regulation. Neuron. 1992 Jan;8(1):135–143. doi: 10.1016/0896-6273(92)90115-t. [DOI] [PubMed] [Google Scholar]
  32. Nooney J. M., Peters J. A., Lambert J. J. A patch clamp study of the nicotinic acetylcholine receptor of bovine adrenomedullary chromaffin cells in culture. J Physiol. 1992 Sep;455:503–527. doi: 10.1113/jphysiol.1992.sp019314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Papke R. L. The kinetic properties of neuronal nicotinic receptors: genetic basis of functional diversity. Prog Neurobiol. 1993 Oct;41(4):509–531. doi: 10.1016/0301-0082(93)90028-q. [DOI] [PubMed] [Google Scholar]
  34. Sanchez J. A., Dani J. A., Siemen D., Hille B. Slow permeation of organic cations in acetylcholine receptor channels. J Gen Physiol. 1986 Jun;87(6):985–1001. doi: 10.1085/jgp.87.6.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sands S. B., Barish M. E. Calcium permeability of neuronal nicotinic acetylcholine receptor channels in PC12 cells. Brain Res. 1991 Sep 27;560(1-2):38–42. doi: 10.1016/0006-8993(91)91211-i. [DOI] [PubMed] [Google Scholar]
  36. Sargent P. B. The diversity of neuronal nicotinic acetylcholine receptors. Annu Rev Neurosci. 1993;16:403–443. doi: 10.1146/annurev.ne.16.030193.002155. [DOI] [PubMed] [Google Scholar]
  37. Trouslard J., Marsh S. J., Brown D. A. Calcium entry through nicotinic receptor channels and calcium channels in cultured rat superior cervical ganglion cells. J Physiol. 1993 Aug;468:53–71. doi: 10.1113/jphysiol.1993.sp019759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vernino S., Amador M., Luetje C. W., Patrick J., Dani J. A. Calcium modulation and high calcium permeability of neuronal nicotinic acetylcholine receptors. Neuron. 1992 Jan;8(1):127–134. doi: 10.1016/0896-6273(92)90114-s. [DOI] [PubMed] [Google Scholar]
  39. Vernino S., Rogers M., Radcliffe K. A., Dani J. A. Quantitative measurement of calcium flux through muscle and neuronal nicotinic acetylcholine receptors. J Neurosci. 1994 Sep;14(9):5514–5524. doi: 10.1523/JNEUROSCI.14-09-05514.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Villarroel A., Herlitze S., Koenen M., Sakmann B. Location of a threonine residue in the alpha-subunit M2 transmembrane segment that determines the ion flow through the acetylcholine receptor channel. Proc Biol Sci. 1991 Jan 22;243(1306):69–74. doi: 10.1098/rspb.1991.0012. [DOI] [PubMed] [Google Scholar]
  41. Vyklicky L., Jr, Krusek J., Edwards C. Differences in the pore sizes of the N-methyl-D-aspartate and kainate cation channels. Neurosci Lett. 1988 Jul 8;89(3):313–318. doi: 10.1016/0304-3940(88)90545-9. [DOI] [PubMed] [Google Scholar]
  42. Westbrook G. L., Mayer M. L. Micromolar concentrations of Zn2+ antagonize NMDA and GABA responses of hippocampal neurons. Nature. 1987 Aug 13;328(6131):640–643. doi: 10.1038/328640a0. [DOI] [PubMed] [Google Scholar]
  43. Woodhull A. M. Ionic blockage of sodium channels in nerve. J Gen Physiol. 1973 Jun;61(6):687–708. doi: 10.1085/jgp.61.6.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yang J. Ion permeation through 5-hydroxytryptamine-gated channels in neuroblastoma N18 cells. J Gen Physiol. 1990 Dec;96(6):1177–1198. doi: 10.1085/jgp.96.6.1177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zhorov B. S., Brovtsyna N. B., Gmiro V. E., Lukomskaya NYa, Serdyuk S. E., Potapyeva N. N., Magazanik L. G., Kurenniy D. E., Skok V. I. Dimensions of the ion channel in neuronal nicotinic acetylcholine receptor as estimated from analysis of conformation-activity relationships of open-channel blocking drugs. J Membr Biol. 1991 Apr;121(2):119–132. doi: 10.1007/BF01870527. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of General Physiology are provided here courtesy of The Rockefeller University Press

RESOURCES