Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 May;83(9):3022–3026. doi: 10.1073/pnas.83.9.3022

Acetylcholine raises excitability by inhibiting the fast transient potassium current in cultured hippocampal neurons.

Y Nakajima, S Nakajima, R J Leonard, K Yamaguchi
PMCID: PMC323439  PMID: 3010326

Abstract

The effects of acetylcholine on cultured hippocampal neurons were investigated by using the whole-cell version of the patch-clamp technique. The CA1 region of the hippocampus was excised from brain slices of young rats (12-19 day old), incubated in a papain solution, and dissociated. Neurons were plated on a glial feeder layer. The experiments were conducted mostly on neurons cultured for 2-6 days. Upon depolarization under voltage clamp, these cells exhibited a fast transient outward current (A-current), which was inhibited by 4-aminopyridine (2.5 mM). Acetylcholine (0.1 microM) also inhibited this A-current, as did the muscarinic agonists bethanechol and muscarine. As expected from their inhibition of the A-current, acetylcholine and 4-aminopyridine both increased the amplitude of the action potential and prolonged its duration. We conclude that the inhibition of the A-current constitutes a mechanism by which acetylcholine exerts its excitatory influence on hippocampal neurons.

Full text

PDF
3022

Selected References

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

  1. Aghajanian G. K. Modulation of a transient outward current in serotonergic neurones by alpha 1-adrenoceptors. Nature. 1985 Jun 6;315(6019):501–503. doi: 10.1038/315501a0. [DOI] [PubMed] [Google Scholar]
  2. Akasu T., Koketsu K. Modulation of voltage-dependent currents by muscarinic receptor in sympathetic neurones of bullfrog. Neurosci Lett. 1982 Mar 17;29(1):41–45. doi: 10.1016/0304-3940(82)90361-5. [DOI] [PubMed] [Google Scholar]
  3. Barker J. L., Dufy B., Owen D. G., Segal M. Excitable membrane properties of cultured central nervous system neurons and clonal pituitary cells. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 1):259–268. doi: 10.1101/sqb.1983.048.01.028. [DOI] [PubMed] [Google Scholar]
  4. Belluzzi O., Sacchi O., Wanke E. A fast transient outward current in the rat sympathetic neurone studied under voltage-clamp conditions. J Physiol. 1985 Jan;358:91–108. doi: 10.1113/jphysiol.1985.sp015542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown D. A., Adams P. R. Muscarinic suppression of a novel voltage-sensitive K+ current in a vertebrate neurone. Nature. 1980 Feb 14;283(5748):673–676. doi: 10.1038/283673a0. [DOI] [PubMed] [Google Scholar]
  6. Choi D. W., Fischbach G. D. GABA conductance of chick spinal cord and dorsal root ganglion neurons in cell culture. J Neurophysiol. 1981 Apr;45(4):605–620. doi: 10.1152/jn.1981.45.4.605. [DOI] [PubMed] [Google Scholar]
  7. Cole A. E., Nicoll R. A. Characterization of a slow cholinergic post-synaptic potential recorded in vitro from rat hippocampal pyramidal cells. J Physiol. 1984 Jul;352:173–188. doi: 10.1113/jphysiol.1984.sp015285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Connor J. A., Stevens C. F. Prediction of repetitive firing behaviour from voltage clamp data on an isolated neurone soma. J Physiol. 1971 Feb;213(1):31–53. doi: 10.1113/jphysiol.1971.sp009366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Connor J. A., Stevens C. F. Voltage clamp studies of a transient outward membrane current in gastropod neural somata. J Physiol. 1971 Feb;213(1):21–30. doi: 10.1113/jphysiol.1971.sp009365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Connor J. A., Walter D., McKown R. Neural repetitive firing: modifications of the Hodgkin-Huxley axon suggested by experimental results from crustacean axons. Biophys J. 1977 Apr;18(1):81–102. doi: 10.1016/S0006-3495(77)85598-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dichter M. A. Rat cortical neurons in cell culture: culture methods, cell morphology, electrophysiology, and synapse formation. Brain Res. 1978 Jun 30;149(2):279–293. doi: 10.1016/0006-8993(78)90476-6. [DOI] [PubMed] [Google Scholar]
  12. Galvan M., Sedlmeir C. Outward currents in voltage-clamped rat sympathetic neurones. J Physiol. 1984 Nov;356:115–133. doi: 10.1113/jphysiol.1984.sp015456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gustafsson B., Galvan M., Grafe P., Wigström H. A transient outward current in a mammalian central neurone blocked by 4-aminopyridine. Nature. 1982 Sep 16;299(5880):252–254. doi: 10.1038/299252a0. [DOI] [PubMed] [Google Scholar]
  14. HAGIWARA S., KUSANO K., SAITO N. Membrane changes of Onchidium nerve cell in potassium-rich media. J Physiol. 1961 Mar;155:470–489. doi: 10.1113/jphysiol.1961.sp006640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. HODGKIN A. L., HUXLEY A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol. 1952 Aug;117(4):500–544. doi: 10.1113/jphysiol.1952.sp004764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Halliwell J. V., Adams P. R. Voltage-clamp analysis of muscarinic excitation in hippocampal neurons. Brain Res. 1982 Oct 28;250(1):71–92. doi: 10.1016/0006-8993(82)90954-4. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Heuser J. E., Reese T. S., Dennis M. J., Jan Y., Jan L., Evans L. Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release. J Cell Biol. 1979 May;81(2):275–300. doi: 10.1083/jcb.81.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kaczmarek L. K., Strumwasser F. A voltage-clamp analysis of currents underlying cyclic AMP-induced membrane modulation in isolated peptidergic neurons of Aplysia. J Neurophysiol. 1984 Aug;52(2):340–349. doi: 10.1152/jn.1984.52.2.340. [DOI] [PubMed] [Google Scholar]
  20. Katz B., Miledi R. A study of synaptic transmission in the absence of nerve impulses. J Physiol. 1967 Sep;192(2):407–436. doi: 10.1113/jphysiol.1967.sp008307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krnjević K., Pumain R., Renaud L. The mechanism of excitation by acetylcholine in the cerebral cortex. J Physiol. 1971 May;215(1):247–268. doi: 10.1113/jphysiol.1971.sp009467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakajima S. Analysis of K inactivation and TEA action in the supramedullary cells of puffer. J Gen Physiol. 1966 Mar;49(4):629–640. doi: 10.1085/jgp.49.4.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nakajima S., Kusano K. Behavior of delayed current under voltage clamp in the supramedullary neurons of puffer. J Gen Physiol. 1966 Mar;49(4):613–628. doi: 10.1085/jgp.49.4.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nakajima Y., Nakajima S., Obata K., Carlson C. G., Yamaguchi K. Dissociated cell culture of cholinergic neurons from nucleus basalis of Meynert and other basal forebrain nuclei. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6325–6329. doi: 10.1073/pnas.82.18.6325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Neher E. Two fast transient current components during voltage clamp on snail neurons. J Gen Physiol. 1971 Jul;58(1):36–53. doi: 10.1085/jgp.58.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nowak L. M., Macdonald R. L. Ionic mechanism of muscarinic cholinergic depolarization of mouse spinal cord neurons in cell culture. J Neurophysiol. 1983 Mar;49(3):792–803. doi: 10.1152/jn.1983.49.3.792. [DOI] [PubMed] [Google Scholar]
  27. Raiteri M., Leardi R., Marchi M. Heterogeneity of presynaptic muscarinic receptors regulating neurotransmitter release in the rat brain. J Pharmacol Exp Ther. 1984 Jan;228(1):209–214. [PubMed] [Google Scholar]
  28. Segal M., Rogawski M. A., Barker J. L. A transient potassium conductance regulates the excitability of cultured hippocampal and spinal neurons. J Neurosci. 1984 Feb;4(2):604–609. doi: 10.1523/JNEUROSCI.04-02-00604.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shimahara T. Presynaptic modulation of transmitter release by the early outward potassium current in Aplysia. Brain Res. 1983 Mar 14;263(1):51–56. doi: 10.1016/0006-8993(83)91199-x. [DOI] [PubMed] [Google Scholar]
  30. Strong J. A. Modulation of potassium current kinetics in bag cell neurons of Aplysia by an activator of adenylate cyclase. J Neurosci. 1984 Nov;4(11):2772–2783. doi: 10.1523/JNEUROSCI.04-11-02772.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. TAKEUCHI A., TAKEUCHI N. Electrical changes in pre- and postsynaptic axons of the giant synapse of Loligo. J Gen Physiol. 1962 Jul;45:1181–1193. doi: 10.1085/jgp.45.6.1181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thompson S. H. Three pharmacologically distinct potassium channels in molluscan neurones. J Physiol. 1977 Feb;265(2):465–488. doi: 10.1113/jphysiol.1977.sp011725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zbicz K. L., Weight F. F. Transient voltage and calcium-dependent outward currents in hippocampal CA3 pyramidal neurons. J Neurophysiol. 1985 Apr;53(4):1038–1058. doi: 10.1152/jn.1985.53.4.1038. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES