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. 1994 Aug;112(4):1160–1166. doi: 10.1111/j.1476-5381.1994.tb13205.x

Muscarinic acetylcholine response in pyramidal neurones of rat cerebral cortex.

M Nishikawa 1, M Munakata 1, N Akaike 1
PMCID: PMC1910245  PMID: 7952877

Abstract

1. The effects of acetylcholine (ACh) on pyramidal neurons acutely dissociated from the rat cerebral cortex were studied in the whole-cell mode, by use of the nystatin-perforated patch recording configuration. 2. ACh induced a net inward current (IACh) accompanied by a membrane conductance decrease at a holding potential (VH) of -40 mV. IACh increased in a concentration-dependent manner with a half-maximum concentration (EC50) of 8.7 x 10(-7) M. 3. IACh mainly resulted from the suppression of the voltage- and time-dependent K+ current (M-current). 4. Muscarine and muscarinic agonists such as McN-A-343, oxotremorine and oxotremorine-M mimicked the ACh response. The potency was in the order of oxotremorine-M > McN-A-343 > or = muscarine > oxotremorine. 5. Pirenzepine shifted the concentration-response curve for ACh to the right and the corresponding Schild plot yielded a pA2 value of 7.81. Other muscarinic antagonists also reversibly blocked IACh in a concentration-dependent manner. The inhibitory potency was in the order of atropine > 4-DAMP > pirenzepine > AF-DX-116. 6. IACh could be induced normally even after pre-incubation of dissociated neurones in external solution with 200 ng ml-1 pertussis toxin (PTX) for 8 h, whereas the inhibitory effect of ACh on high-voltage-activated Ca2+ channels was completely abolished by the PTX treatment.

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Selected References

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  1. ARUNLAKSHANA O., SCHILD H. O. Some quantitative uses of drug antagonists. Br J Pharmacol Chemother. 1959 Mar;14(1):48–58. doi: 10.1111/j.1476-5381.1959.tb00928.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Akaike N., Kostyuk P. G., Osipchuk Y. V. Dihydropyridine-sensitive low-threshold calcium channels in isolated rat hypothalamic neurones. J Physiol. 1989 May;412:181–195. doi: 10.1113/jphysiol.1989.sp017610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Akaike N., Lee K. S., Brown A. M. The calcium current of Helix neuron. J Gen Physiol. 1978 May;71(5):509–531. doi: 10.1085/jgp.71.5.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benardo L. S., Prince D. A. Cholinergic excitation of mammalian hippocampal pyramidal cells. Brain Res. 1982 Oct 14;249(2):315–331. doi: 10.1016/0006-8993(82)90066-x. [DOI] [PubMed] [Google Scholar]
  5. Bernheim L., Mathie A., Hille B. Characterization of muscarinic receptor subtypes inhibiting Ca2+ current and M current in rat sympathetic neurons. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9544–9548. doi: 10.1073/pnas.89.20.9544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Birdsall N. J., Burgen A. S., Hulme E. C., Stockton J. M., Zigmond M. J. The effect of McN-A-343 on muscarinic receptors in the cerebral cortex and heart. Br J Pharmacol. 1983 Feb;78(2):257–259. doi: 10.1111/j.1476-5381.1983.tb09388.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bonner T. I. The molecular basis of muscarinic receptor diversity. Trends Neurosci. 1989 Apr;12(4):148–151. doi: 10.1016/0166-2236(89)90054-4. [DOI] [PubMed] [Google Scholar]
  8. Bonner T. I., Young A. C., Brann M. R., Buckley N. J. Cloning and expression of the human and rat m5 muscarinic acetylcholine receptor genes. Neuron. 1988 Jul;1(5):403–410. doi: 10.1016/0896-6273(88)90190-0. [DOI] [PubMed] [Google Scholar]
  9. Brann M. R., Buckley N. J., Jones S. V., Bonner T. I. Expression of a cloned muscarinic receptor in A9 L cells. Mol Pharmacol. 1987 Oct;32(4):450–455. [PubMed] [Google Scholar]
  10. 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]
  11. Brown D. A., Marrion N. V., Smart T. G. On the transduction mechanism for muscarine-induced inhibition of M-current in cultured rat sympathetic neurones. J Physiol. 1989 Jun;413:469–488. doi: 10.1113/jphysiol.1989.sp017664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Buckley N. J., Bonner T. I., Brann M. R. Localization of a family of muscarinic receptor mRNAs in rat brain. J Neurosci. 1988 Dec;8(12):4646–4652. doi: 10.1523/JNEUROSCI.08-12-04646.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Buckley N. J., Bonner T. I., Buckley C. M., Brann M. R. Antagonist binding properties of five cloned muscarinic receptors expressed in CHO-K1 cells. Mol Pharmacol. 1989 Apr;35(4):469–476. [PubMed] [Google Scholar]
  14. Constanti A., Brown D. A. M-Currents in voltage-clamped mammalian sympathetic neurones. Neurosci Lett. 1981 Jul 17;24(3):289–294. doi: 10.1016/0304-3940(81)90173-7. [DOI] [PubMed] [Google Scholar]
  15. Constanti A., Sim J. A. Muscarinic receptors mediating suppression of the M-current in guinea-pig olfactory cortex neurones may be of the M2-subtype. Br J Pharmacol. 1987 Jan;90(1):3–5. doi: 10.1111/j.1476-5381.1987.tb16818.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Coyle J. T., Price D. L., DeLong M. R. Alzheimer's disease: a disorder of cortical cholinergic innervation. Science. 1983 Mar 11;219(4589):1184–1190. doi: 10.1126/science.6338589. [DOI] [PubMed] [Google Scholar]
  17. Downes C. P. Receptor-stimulated inositol phospholipid metabolism in the central nervous system. Cell Calcium. 1982 Oct;3(4-5):413–428. doi: 10.1016/0143-4160(82)90027-6. [DOI] [PubMed] [Google Scholar]
  18. Dutar P., Nicoll R. A. Classification of muscarinic responses in hippocampus in terms of receptor subtypes and second-messenger systems: electrophysiological studies in vitro. J Neurosci. 1988 Nov;8(11):4214–4224. doi: 10.1523/JNEUROSCI.08-11-04214.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Freedman S. B., Beer M. S., Harley E. A. Muscarinic M1, M2 receptor binding. Relationship with functional efficacy. Eur J Pharmacol. 1988 Oct 26;156(1):133–142. doi: 10.1016/0014-2999(88)90155-0. [DOI] [PubMed] [Google Scholar]
  20. Fukuda K., Higashida H., Kubo T., Maeda A., Akiba I., Bujo H., Mishina M., Numa S. Selective coupling with K+ currents of muscarinic acetylcholine receptor subtypes in NG108-15 cells. Nature. 1988 Sep 22;335(6188):355–358. doi: 10.1038/335355a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  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. Hulme E. C., Birdsall N. J., Buckley N. J. Muscarinic receptor subtypes. Annu Rev Pharmacol Toxicol. 1990;30:633–673. doi: 10.1146/annurev.pa.30.040190.003221. [DOI] [PubMed] [Google Scholar]
  24. Ito C., Wakamori M., Akaike N. Dual effect of glycine on isolated rat suprachiasmatic neurons. Am J Physiol. 1991 Feb;260(2 Pt 1):C213–C218. doi: 10.1152/ajpcell.1991.260.2.C213. [DOI] [PubMed] [Google Scholar]
  25. Kijima H., Kijima S. Cooperative response of chemically excitable membrane. II. Two-state models and their limitations. J Theor Biol. 1980 Feb 7;82(3):425–463. doi: 10.1016/0022-5193(80)90248-9. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Kurose H., Katada T., Amano T., Ui M. Specific uncoupling by islet-activating protein, pertussis toxin, of negative signal transduction via alpha-adrenergic, cholinergic, and opiate receptors in neuroblastoma x glioma hybrid cells. J Biol Chem. 1983 Apr 25;258(8):4870–4875. [PubMed] [Google Scholar]
  28. Lambrecht G., Moser U., Grimm U., Pfaff O., Hermanni U., Hildebrandt C., Waelbroeck M., Christophe J., Mutschler E. New functionally selective muscarinic agonists. Life Sci. 1993;52(5-6):481–488. doi: 10.1016/0024-3205(93)90305-m. [DOI] [PubMed] [Google Scholar]
  29. Lazareno S., Buckley N. J., Roberts F. F. Characterization of muscarinic M4 binding sites in rabbit lung, chicken heart, and NG108-15 cells. Mol Pharmacol. 1990 Dec;38(6):805–815. [PubMed] [Google Scholar]
  30. Levey A. I., Kitt C. A., Simonds W. F., Price D. L., Brann M. R. Identification and localization of muscarinic acetylcholine receptor proteins in brain with subtype-specific antibodies. J Neurosci. 1991 Oct;11(10):3218–3226. doi: 10.1523/JNEUROSCI.11-10-03218.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Marrion N. V., Smart T. G., Marsh S. J., Brown D. A. Muscarinic suppression of the M-current in the rat sympathetic ganglion is mediated by receptors of the M1-subtype. Br J Pharmacol. 1989 Oct;98(2):557–573. doi: 10.1111/j.1476-5381.1989.tb12630.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McCormick D. A., Prince D. A. Two types of muscarinic response to acetylcholine in mammalian cortical neurons. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6344–6348. doi: 10.1073/pnas.82.18.6344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McKinney M., Richelson E. The coupling of the neuronal muscarinic receptor to responses. Annu Rev Pharmacol Toxicol. 1984;24:121–146. doi: 10.1146/annurev.pa.24.040184.001005. [DOI] [PubMed] [Google Scholar]
  34. Messer W. S., Jr, Bohnett M., Stibbe J. Evidence for a preferential involvement of M1 muscarinic receptors in representational memory. Neurosci Lett. 1990 Aug 14;116(1-2):184–189. doi: 10.1016/0304-3940(90)90407-z. [DOI] [PubMed] [Google Scholar]
  35. Mesulam M. M., Geula C. Nucleus basalis (Ch4) and cortical cholinergic innervation in the human brain: observations based on the distribution of acetylcholinesterase and choline acetyltransferase. J Comp Neurol. 1988 Sep 8;275(2):216–240. doi: 10.1002/cne.902750205. [DOI] [PubMed] [Google Scholar]
  36. Micheletti R., Montagna E., Giachetti A. AF-DX 116, a cardioselective muscarinic antagonist. J Pharmacol Exp Ther. 1987 May;241(2):628–634. [PubMed] [Google Scholar]
  37. Murase K., Randic M., Shirasaki T., Nakagawa T., Akaike N. Serotonin suppresses N-methyl-D-aspartate responses in acutely isolated spinal dorsal horn neurons of the rat. Brain Res. 1990 Aug 13;525(1):84–91. doi: 10.1016/0006-8993(90)91323-9. [DOI] [PubMed] [Google Scholar]
  38. Murayama T., Ui M. Loss of the inhibitory function of the guanine nucleotide regulatory component of adenylate cyclase due to its ADP ribosylation by islet-activating protein, pertussis toxin, in adipocyte membranes. J Biol Chem. 1983 Mar 10;258(5):3319–3326. [PubMed] [Google Scholar]
  39. Pfaffinger P. Muscarine and t-LHRH suppress M-current by activating an IAP-insensitive G-protein. J Neurosci. 1988 Sep;8(9):3343–3353. doi: 10.1523/JNEUROSCI.08-09-03343.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Robbins J., Caulfield M. P., Higashida H., Brown D. A. Genotypic m3-Muscarinic Receptors Preferentially Inhibit M-currents in DNA-transfected NG108-15 Neuroblastoma x Glioma Hybrid Cells. Eur J Neurosci. 1991;3(8):820–824. doi: 10.1111/j.1460-9568.1991.tb01678.x. [DOI] [PubMed] [Google Scholar]
  41. Shirasaki T., Harata N., Akaike N. Metabotropic glutamate response in acutely dissociated hippocampal CA1 pyramidal neurones of the rat. J Physiol. 1994 Mar 15;475(3):439–453. doi: 10.1113/jphysiol.1994.sp020084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Squire L. R., Davis H. P. The pharmacology of memory: a neurobiological perspective. Annu Rev Pharmacol Toxicol. 1981;21:323–356. doi: 10.1146/annurev.pa.21.040181.001543. [DOI] [PubMed] [Google Scholar]
  43. Uneyama H., Ueno S., Akaike N. Serotonin-operated potassium current in CA1 neurons dissociated from rat hippocampus. J Neurophysiol. 1993 Apr;69(4):1044–1052. doi: 10.1152/jn.1993.69.4.1044. [DOI] [PubMed] [Google Scholar]
  44. Wakamori M., Hidaka H., Akaike N. Hyperpolarizing muscarinic responses of freshly dissociated rat hippocampal CA1 neurones. J Physiol. 1993 Apr;463:585–604. doi: 10.1113/jphysiol.1993.sp019612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wall S. J., Yasuda R. P., Hory F., Flagg S., Martin B. M., Ginns E. I., Wolfe B. B. Production of antisera selective for m1 muscarinic receptors using fusion proteins: distribution of m1 receptors in rat brain. Mol Pharmacol. 1991 May;39(5):643–649. [PubMed] [Google Scholar]
  46. Womble M. D., Moises H. C. Muscarinic inhibition of M-current and a potassium leak conductance in neurones of the rat basolateral amygdala. J Physiol. 1992 Nov;457:93–114. doi: 10.1113/jphysiol.1992.sp019366. [DOI] [PMC free article] [PubMed] [Google Scholar]

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