Skip to main content
NCBI home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1991 Sep;104(1):39–44. doi: 10.1111/j.1476-5381.1991.tb12381.x

Pharmacology of the putative M4 muscarinic receptor mediating Ca-current inhibition in neuroblastoma x glioma hybrid (NG 108-15) cells.

M P Caulfield 1, D A Brown 1
PMCID: PMC1908258  PMID: 1786516

Abstract

1. We have assessed the potency of a range of agonists and antagonists on the muscarinic receptor responsible for inhibiting the Ca-current (ICa) in NG 108-15 hybrid cells. 2. Acetylcholine (ACh), oxotremorine-M and carbachol were potent 'full' agonists (EC50 values were 0.11 microM, 0.14 microM and 2 microM, respectively). Maximum inhibition of peak high-threshold ICa by these agonists was 39.5%. (+/-)-Muscarine, methylfurmethide and arecaidine propargyl ester (APE) were 'partial' agonists, with EC50 values of 0.54 microM, 0.84 microM and 0.1 microM, respectively. 3. Atropine, pirenzepine and himbacine were potent antagonists of muscarinic inhibition of ICa, with apparent pKB values of 9.8, 7.74 and 8.83, respectively. Methoctramine was relatively weak (pKB = 7.63). Atropine and pirenzepine depressed maximum responses to agonists, probably because these antagonists have relatively slow dissociation rates. 4. The characteristic pharmacological profile found for the M4 receptors in these functional experiments (himbacine high affinity, pirenzepine moderate to high affinity, methoctramine low affinity) corresponds well with data from earlier binding experiments (Lazareno et al., 1990). Since mRNA hybridising to probes for the m4 receptor genotype can be detected in these cells, it is suggested that these pharmacological characteristics identify the equivalent expressed receptor subtype M4.

Full text

PDF

Selected References

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

  1. Anwar-ul S., Gilani H., Cobbin L. B. The cardio-selectivity of himbacine: a muscarine receptor antagonist. Naunyn Schmiedebergs Arch Pharmacol. 1986 Jan;332(1):16–20. doi: 10.1007/BF00633191. [DOI] [PubMed] [Google Scholar]
  2. Barlow R. B., McMillen L. S., Veale M. A. The use of 4-diphenylacetoxy-N-(2-chloroethyl)-piperidine (4-DAMP mustard) for estimating the apparent affinities of some agonists acting at muscarinic receptors in guinea-pig ileum. Br J Pharmacol. 1991 Mar;102(3):657–662. doi: 10.1111/j.1476-5381.1991.tb12229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baumgold J., Drobnick A. An agonist that is selective for adenylate cyclase-coupled muscarinic receptors. Mol Pharmacol. 1989 Sep;36(3):465–470. [PubMed] [Google Scholar]
  4. Bonner T. I., Buckley N. J., Young A. C., Brann M. R. Identification of a family of muscarinic acetylcholine receptor genes. Science. 1987 Jul 31;237(4814):527–532. doi: 10.1126/science.3037705. [DOI] [PubMed] [Google Scholar]
  5. Brown D. A., Forward A., Marsh S. Antagonist discrimination between ganglionic and ileal muscarinic receptors. Br J Pharmacol. 1980;71(2):362–364. doi: 10.1111/j.1476-5381.1980.tb10948.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown D. A., Higashida H. Voltage- and calcium-activated potassium currents in mouse neuroblastoma x rat glioma hybrid cells. J Physiol. 1988 Mar;397:149–165. doi: 10.1113/jphysiol.1988.sp016993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Choo L. K., Mitchelson F., Vong Y. M. Interaction of some muscarinic agonists and antagonists at the prejunctional muscarinic receptor in the rabbit ear artery preparation. Br J Pharmacol. 1986 Apr;87(4):733–740. doi: 10.1111/j.1476-5381.1986.tb14591.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Docherty R. J. Gadolinium selectively blocks a component of calcium current in rodent neuroblastoma x glioma hybrid (NG108-15) cells. J Physiol. 1988 Apr;398:33–47. doi: 10.1113/jphysiol.1988.sp017027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Docherty R. J., McFadzean I. Noradrenaline-Induced Inhibition of Voltage-Sensitive Calcium Currents in NG108-15 Hybrid Cells. Eur J Neurosci. 1989 Mar;1(2):132–140. doi: 10.1111/j.1460-9568.1989.tb00780.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Eltze M. Muscarinic M1- and M2-receptors mediating opposite effects on neuromuscular transmission in rabbit vas deferens. Eur J Pharmacol. 1988 Jul 7;151(2):205–221. doi: 10.1016/0014-2999(88)90801-1. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Gähwiler B. H., Brown D. A. Muscarine affects calcium-currents in rat hippocampal pyramidal cells in vitro. Neurosci Lett. 1987 May 19;76(3):301–306. doi: 10.1016/0304-3940(87)90419-8. [DOI] [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. Hasuo H., Gallagher J. P., Shinnick-Gallagher P. Disinhibition in the rat septum mediated by M1 muscarinic receptors. Brain Res. 1988 Jan 12;438(1-2):323–327. doi: 10.1016/0006-8993(88)91356-x. [DOI] [PubMed] [Google Scholar]
  17. Higashida H., Hashii M., Fukuda K., Caulfield M. P., Numa S., Brown D. A. Selective coupling of different muscarinic acetylcholine receptors to neuronal calcium currents in DNA-transfected cells. Proc Biol Sci. 1990 Oct 22;242(1303):68–74. doi: 10.1098/rspb.1990.0105. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Jeck D., Lindmar R., Löffelholz K., Wanke M. Subtypes of muscarinic receptor on cholinergic nerves and atrial cells of chicken and guinea-pig hearts. Br J Pharmacol. 1988 Feb;93(2):357–366. doi: 10.1111/j.1476-5381.1988.tb11442.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kubo T., Fukuda K., Mikami A., Maeda A., Takahashi H., Mishina M., Haga T., Haga K., Ichiyama A., Kangawa K. Cloning, sequencing and expression of complementary DNA encoding the muscarinic acetylcholine receptor. Nature. 1986 Oct 2;323(6087):411–416. doi: 10.1038/323411a0. [DOI] [PubMed] [Google Scholar]
  21. Kubo T., Maeda A., Sugimoto K., Akiba I., Mikami A., Takahashi H., Haga T., Haga K., Ichiyama A., Kangawa K. Primary structure of porcine cardiac muscarinic acetylcholine receptor deduced from the cDNA sequence. FEBS Lett. 1986 Dec 15;209(2):367–372. doi: 10.1016/0014-5793(86)81144-9. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lazareno S., Roberts F. F. Functional and binding studies with muscarinic M2-subtype selective antagonists. Br J Pharmacol. 1989 Sep;98(1):309–317. doi: 10.1111/j.1476-5381.1989.tb16896.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McFadzean I., Mullaney I., Brown D. A., Milligan G. Antibodies to the GTP binding protein, Go, antagonize noradrenaline-induced calcium current inhibition in NG108-15 hybrid cells. Neuron. 1989 Aug;3(2):177–182. doi: 10.1016/0896-6273(89)90030-5. [DOI] [PubMed] [Google Scholar]
  25. McGee R., Simpson P., Christian C., Mata M., Nelson P., Nirenberg M. Regulation of acetylcholine release from neuroblastoma x glioma hybrid cells. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1314–1318. doi: 10.1073/pnas.75.3.1314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Michel A. D., Delmendo R., Stefanich E., Whiting R. L. Binding characteristics of the muscarinic receptor subtype of the NG108-15 cell line. Naunyn Schmiedebergs Arch Pharmacol. 1989 Jul;340(1):62–67. doi: 10.1007/BF00169208. [DOI] [PubMed] [Google Scholar]
  27. Miller R. J. Receptor-mediated regulation of calcium channels and neurotransmitter release. FASEB J. 1990 Dec;4(15):3291–3299. [PubMed] [Google Scholar]
  28. Mochida S., Kobayashi H. Activation of M2 muscarinic receptors causes an alteration of action potentials by modulation of Ca entry in isolated sympathetic neurons of rabbits. Neurosci Lett. 1986 Dec 12;72(2):199–204. doi: 10.1016/0304-3940(86)90080-7. [DOI] [PubMed] [Google Scholar]
  29. Peralta E. G., Ashkenazi A., Winslow J. W., Smith D. H., Ramachandran J., Capon D. J. Distinct primary structures, ligand-binding properties and tissue-specific expression of four human muscarinic acetylcholine receptors. EMBO J. 1987 Dec 20;6(13):3923–3929. doi: 10.1002/j.1460-2075.1987.tb02733.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Robbins J., Sim J. A. A transient outward current in NG108-15 neuroblastoma x glioma hybrid cells. Pflugers Arch. 1990 Apr;416(1-2):130–137. doi: 10.1007/BF00370234. [DOI] [PubMed] [Google Scholar]
  31. Sheridan R. D., Sutor B. Presynaptic M1 muscarinic cholinoceptors mediate inhibition of excitatory synaptic transmission in the hippocampus in vitro. Neurosci Lett. 1990 Jan 22;108(3):273–278. doi: 10.1016/0304-3940(90)90653-q. [DOI] [PubMed] [Google Scholar]
  32. Song S. Y., Saito K., Noguchi K., Konishi S. Different GTP-binding proteins mediate regulation of calcium channels by acetylcholine and noradrenaline in rat sympathetic neurons. Brain Res. 1989 Aug 14;494(2):383–386. doi: 10.1016/0006-8993(89)90609-4. [DOI] [PubMed] [Google Scholar]
  33. Tse A., Clark R. B., Giles W. R. Muscarinic modulation of calcium current in neurones from the interatrial septum of bull-frog heart. J Physiol. 1990 Aug;427:127–149. doi: 10.1113/jphysiol.1990.sp018164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wanke E., Ferroni A., Malgaroli A., Ambrosini A., Pozzan T., Meldolesi J. Activation of a muscarinic receptor selectively inhibits a rapidly inactivated Ca2+ current in rat sympathetic neurons. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4313–4317. doi: 10.1073/pnas.84.12.4313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Williams S. H., Constanti A. A quantitative study of the effects of some muscarinic antagonists on the guinea-pig olfactory cortex slice. Br J Pharmacol. 1988 Apr;93(4):855–862. doi: 10.1111/j.1476-5381.1988.tb11472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES