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Pharmacological characterization of a nicotinic autoreceptor in rat hippocampal synaptosomes

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Abstract

The modulation of [3H]ACh release by nicotinic compounds was studied in superfused rat hippocampal synaptosomes loaded with [3H]choline. (−)-Nicotine (0.1–10 μM) evoked a dose-dependent increase in [3H]ACh release; higher concentrations were less effective. Nicotine-evoked release was Ca2+-dependent, and blocked by the nicotinic antagonists dihydro-β-erythroidine, mecamylamine, and pempidine. The α7-selective antagonist methyllycaconitine did not inhibit nicotine-evoked release when tested at 1 μM, although at 10 μM some attenuation of the response was observed. Six agonists tested were equally efficacious in stimulating [3H]ACh release, as judged by the maximum responses, and gave the following EC50 values: (±)-epibatidine 0.12 μM; (+)-anatoxin-a 0.14 μM; (−)-nicotine 0.99 μM; (−)-cytisine 1.06 μM; ABT-418 2.6 μM; isoarecolone 43 μM. Each agonist generated a “bell-shaped” dose response curve, suggesting desensitisation at higher concentrations. This is supported by analysis of repetitive stimulation with (−)-nicotine and (−)-cytisine: S2/S1 ratios declined sharply with increasing concentration, whereas subsequent KCl-evoked release remained constant. These results are discussed in terms of possible nicotinic receptor subtypes that might be present on hippocampal nerve terminals.

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Abbreviations

DHβE:

dihydro-β-erythroidine

MLA:

methyllycaconitine

nAChR:

nicotinic acetylcholine receptor

References

  1. Wonnacott, S., Drasdo, A., Sanderson, E., and Rowell, P. 1990. Presynaptic nicotinic receptors and the modulation of transmitter release. Pages 87–101,in Marsh, J. (ed.), Ciba Foundation Symposium 152: The Biology of Nicotine Dependence, John Wiley and Sons, Chichester.

    Google Scholar 

  2. McGehee, D. S., Heath, M. J. S., Gelber, S., Devay, P., and Role, L. W. 1995. Nicotine enhancement of fast excitatory synaptic transmission in CNS by presynaptic receptors. Science 269:1692–1696.

    Article  PubMed  CAS  Google Scholar 

  3. Rapier, C., Lunt, G. G., and Wonnacott, S. 1988. Stereoselective nicotine-induced release of dopamine from striatal synaptosomes: concentration dependence and repetitive stimulation. J. Neurochem. 50:1123–1130.

    Article  PubMed  CAS  Google Scholar 

  4. Rapier, C., Lunt, G. G., and Wonnacott, S. 1990. Nicotinic modulation of [3H]dopamine release from striatal synaptosomes: pharmacological characterisation. J. neurochem. 54:937–945.

    Article  PubMed  CAS  Google Scholar 

  5. Grady, S., Marks, M., Wonnacott, S., and Collins, A. C. 1992. Characterisation of nicotinic receptor mediated [3H]dopamine release from synaptosomes prepared from mouse striatum. J. Neurochem. 59:848–856.

    Article  PubMed  CAS  Google Scholar 

  6. El-Bizri, H. and Clarke, P. B. S. 1994. Blockade of nicotinic receptor-mediated release of dopamine from striatal synaptosomes by chlorisondamine and other nicotinic antagonists administered in vitro. Brit. J. Pharmacol. 111:406–413.

    CAS  Google Scholar 

  7. Soliakov, L., Gallagher, T., and Wonnacott, S. 1995. Anatoxin-a-evoked [3H]dopamine release from rat striatal synaptosomes. Neuropharmacol. 34:1535–1541.

    Article  CAS  Google Scholar 

  8. Soliakov, L. and Wonnacott, S. 1996. Voltage-sensitive Ca2+ channels involved in nicotinic receptor-mediated [3H]dopamine release from rat striatal synaptosomes. J. Neurochem. 67:163–170.

    Article  PubMed  CAS  Google Scholar 

  9. Rapier, C., Wonnacott, S., Lunt, G. G., and Albuquerque, E. X. 1987. The neurotoxin histrionicotoxin interacts with the putative ion channel of the nicotinic acetylcholine receptors in the central nervous system. Febs Lett. 212:292–296.

    Article  PubMed  CAS  Google Scholar 

  10. Drasdo, A., Caulfield, M. P., Bertrand, D., Bertrand, S., and Wonnacott, S. 1992. Methyllycaconitine: a novel nicotinic antagonist. Mol. Cell. Neurosci. 3:237–243.

    Article  CAS  PubMed  Google Scholar 

  11. Sacaan, A. I., Dunlop, J. L., and Lloyd, G. K. 1995. Pharmacological characterisation of neuronal acetylcholine gated ion channel receptor-mediated hippocampal norepinephrine and striatal dopamine release from rat brain slices. J. Pharmacol. Exp. Ther. 274:224–230.

    PubMed  CAS  Google Scholar 

  12. Clarke, P. B. S., and Reuben, M. 1996. Release of [3H]noradrenaline from rat hippocampal synaptosomes by nicotine: mediation by different receptor subtypes from striatal [3H]dopamine release. Brit. J. Pharmacol. 117:595–606.

    CAS  Google Scholar 

  13. Sargent, P. B. 1993. The diversity of neuronal nicotinic acetylcholine receptors. Ann. Rev. Neurosci. 16:403–443.

    Article  PubMed  CAS  Google Scholar 

  14. Wada, E., Wada, K., Boulter, J., Deneris, E., Heinemann, S., Patrick, J., and Swanson, L. W. 1989. Distribution of alpha2, alpha3, alpha4, and beta2 neuronal nicotinic receptor subunit mRNAs in the central nervous system: a hybridisation histochemical study in the rat. J. Comp. Neurol. 284:314–335.

    Article  PubMed  CAS  Google Scholar 

  15. Dineley-Miller, K., and Patrick, J. 1992. Gene transcripts for the nicotinic acetylcholine receptor subunit, beta4, are distributed in multiple areas of the rat central nervous system. Mol. Brain Res. 16:339–344.

    Article  PubMed  CAS  Google Scholar 

  16. Wada, E., McKinnon, D., Heinemann, S., Patrick, J., and Swanson, L. 1990. The distribution of mRNA encoded by a new member of the neuronal nicotinic acetylcholine receptor gene family (α5) in the rat central nervous system. Brain Res. 526:45–53.

    Article  PubMed  CAS  Google Scholar 

  17. Bowman, W. C., Marshall, I. G., Gibb, A. J., and Harborne, A. J. 1988. Feedback control of transmitter release at the neuromuscular junction. Trends Pharmacol. Sci. 9:16–20.

    Article  PubMed  CAS  Google Scholar 

  18. Rowell, P. P. and Winkler, D. L. 1984. Nicotinic stimulation of [3H]acetylcholine release from mouse cerebral cortical synaptosomes. J. Neurochem. 43:1593–1598.

    Article  PubMed  CAS  Google Scholar 

  19. Araujo, D. M., Lapchak, P. A., Collier, B., and Quirion, R. 1988. Characterisation of N-[3H]methylcarbamylcholine binding sites and effect of N-methylcarbamylcholine on acetylcholine release in rat brain. J. Neurochem. 51:292–299.

    Article  PubMed  CAS  Google Scholar 

  20. Whiting, P., Esch, F., Shimasaki, S., and Lindstrom, J. 1987. Neuronal nicotinic acetylcholine receptor β-subunit is coded for by the cDNA clone α4. FEBS Lett. 219:459–463.

    Article  PubMed  CAS  Google Scholar 

  21. Flores, C. M., Rogers, S. W., Pabreza, L. A., Wolfe, B. B., and Kellar, K. J. 1992. A subtype of nicotinic cholinergic receptor in rat brain is composed of α4 and β2 subunits and is upregulated by chronic nicotine treatment. Mol. Pharmacol. 41:31–37.

    PubMed  CAS  Google Scholar 

  22. Araujo, D. M., Lanchak, P. A., Robitaille, Y., Gauthier, S., and Quirion, R. 1988. Differential alteration of various cholinergic markers in cortical and subcortical regions of human brain in Alzheimer's disease. J. Neurochem. 50:1914–1923.

    Article  PubMed  CAS  Google Scholar 

  23. Williams, M., Sullivan, J. P., and Arneric, S. P. 1994. Neuronal nicotinic acetylcholine receptors. Drug News And Perspectives 7: 205–223.

    Google Scholar 

  24. Holladay, M. W., Lebold, S.A., and Nan-Horng, L. 1995. Structure-activity relationships of nicotinic acetylcholine receptor agonists as potential treatments for dementia. Drug Dev. Res. 353:191–213.

    Article  Google Scholar 

  25. Clarke, P. B. S. and Pert, A. 1985. Autoradiographic evidence for nicotine receptors on nigrostriatal and mesolimbic dopaminergic neurons. Brain Res. 348:355–358.

    Article  PubMed  CAS  Google Scholar 

  26. Wilkie, G. I., Stephens, M. W., Hutson, P. J., Whiting, P., and Wonnacott, S. 1993. Hippocampal nicotinic autoreceptors modulate acetylcholine release. Biochem. Soc. Trans. 21:429–431.

    PubMed  CAS  Google Scholar 

  27. Thorne, B., Wonnacott, S., and Dunkley, P. R. 1991. Isolation of hippocampal synaptosomes on Percoll gradients: cholinergic markers and ligand binding sites. J. Neurochem. 56:479–484.

    Article  PubMed  CAS  Google Scholar 

  28. Thorne, B. 1990. Nicotinic regulation of acetylcholine release from rat brain hippocampus. PhD Thesis, University of Bath.

  29. Thorne, B., Irons, J., Lunt, G. G., Wonnacott, S., and Dunkley, P. R. 1988. Comparison of methods for rapid isolation of synaptosomes from brain regions, for uptake and release studies. Biochem. Soc. Trans. 16:309–310.

    CAS  Google Scholar 

  30. Garvey, D. S., Wasicak, J., Decker, M. W., Brioni, J. D., Buckley, M. J., Sullivan, J. P., Carrera, G. M., Holladay, M. H., Arneric, S. P., and Williams, M. 1994. Novel isoxazoles which interact with brain cholinergic channel receptors that have intrinsic cognitive enhancing and anxiolytic properties. J. Med. Chem. 37:1055–1059.

    Article  PubMed  CAS  Google Scholar 

  31. Rowell, P. R., & Hillebrand, J. A. 1994. Characterisation of nicotine-induced desensitisation of evoked dopamine release from rat striatal synaptosomes, J. Neurochem. 63:561–569.

    Article  PubMed  CAS  Google Scholar 

  32. Badio, B., and Daly, J. W. 1994. Epibatidine, a potent analgetic and nicotinic agonist. Mol. Pharmacol. 45:563–569.

    PubMed  CAS  Google Scholar 

  33. Sullivan, J. P., Decker, M. W., Brioni, J. D., Donnelly-Robert, D., Anderson, D. J., Bannon, A. W., Kang, C.-H., Adams, P., Piattoni-Kaplan, M., Buckley, M. J., Gopalakrishnan, Williams, M., and Arneric, S. 1994. (±)-Epibatidine elicits a diversity of in vitro and in vivo effects mediated by nicotinic acetylcholine receptors. J. Pharmacol. Exp. Ther. 271:624–631.

    PubMed  CAS  Google Scholar 

  34. Arneric, S. P., Sullivan, J. P., Briggs, C. A., Donnelly-Roberts, D., Anderson, D. J., Raszkiewicz, J. L., Hughes, M. L., Cadman, E. D., Adams, P., Garvey, D. S., Wasicak, J. T., and Williams, M. 1994. (S)-3-Methyl-5-(1-methyl-2-pyrrolidinyl)isoxazole (ABT-418): A novel cholinergic ligand with cognition-enhancing and anxiolytic activities: I. In vitro characterisation. J. Pharmacol. Exp. Ther. 270: 310–318.

    PubMed  CAS  Google Scholar 

  35. Arneric, S. P., Anderson, D. A., Bannon, A. D., Brioni, J. D., Briggs, C. A., Buccafusco, J., Decker, M. W., Donnelly-Roberts, D., Godalakrishnan, M., Holladay, M. W., and Sullivan, J. P. 1995. Preclinical pharmacology of ABT-418: a prototypical cholinergic channel activator for the treatment of Alzheimer's disease. CNS Drug Rev. 1:1–26.

    Article  CAS  Google Scholar 

  36. Whiteaker, P., Garcha, H. S., Wonnacott, S., and Stolerman, I. P. 1995. Locomotor activation and dopamine release produced by nicotine and isoarecolone in rats. Brit. J. Pharmacol. 116:2097–2105.

    CAS  Google Scholar 

  37. Wong, E. T., Holstad, S. G., Mennerick, S. J., Hong, S. E., Zorumski, C. F., and Isenberg, K. E. 1995. Pharmacological and physiological properties of a putative ganglionic nicotinic receptor, alpha3beta4, expressed in transfected eukaryotic cells. Mol. Brain Res. 28:101–109.

    Article  PubMed  CAS  Google Scholar 

  38. Thomas, P., Stephens, M., Wilkie, G., Amar, M., Lunt, G. G., Whiting, P., Gallagher, T., Pereira, E., Alkondon, M., Albuquerque, E. X., and Wonnacott, S. 1993. (+)-anatoxin-a is a potent agonist at neuronal nicotinic acetylcholine receptors. J. Neurochem. 60:2308–2311.

    Article  PubMed  CAS  Google Scholar 

  39. Vibat, C. R. T., Lasalde, J. A., McNamee, M. G., and Ochoa, E. L. M. 1995. Differential desensitization properties of rat neuronal nicotinic acetylcholine receptor subunit combinations expressed inXenopus oocytes. Cell. Mol. Neurobiol. 15:411–425.

    Article  PubMed  CAS  Google Scholar 

  40. Luetje, C. W., and Patrick, J. 1991. Both α- and β-subunits contribute to the agonist sensitivity of neuronal nicotinic acetylcholine receptors. J. Neurosci. 11:837–845.

    PubMed  CAS  Google Scholar 

  41. Papke, R. and Heinemann, S. 1991. The role of the β4 subunit in determining the kinetic properties of rat neuronal nicotinic acetylcholine α3 receptors. J. Physiol. (London) 440:95–112.

    CAS  Google Scholar 

  42. Marks, M. J., Farnham, D. A., Grady, S. R., and Collins, A. C. 1993. Nicotinic receptor function determined by stimulation of rubidium efflux from mouse brain synaptosomes. J. Pharmacol. Exp. Ther. 264:542–552.

    PubMed  CAS  Google Scholar 

  43. Peng, X., Gerzanich, V., Anand, R., Whiting, P. J., and Lindstrom, J. 1994. Nicotine-induced increase in neuronal nicotinic receptors results from a decrease in the rate of receptor turnover. Mol. Pharmacol. 46:523–530.

    PubMed  CAS  Google Scholar 

  44. Court, J. A., Perry, E. K., Spurden, D., Lloyd, S., Gillespie, J. I., Whiting, P., and Barlow, R. 1994. Comparison of the binding of nicotinic agonists to receptors from human and rat cerebral cortex and from chick brain (α4β2) transfected into mouse fibroblasts with ion channel activity. Brain Res. 667:118–122.

    Article  PubMed  CAS  Google Scholar 

  45. Conroy, W. G., Vernallis, A. B., and Berg, D. K. 1992. The α5 gene product assembles with multiple acetylcholine receptor subunits to form distinctive receptor subtypes in brain. Neuron 9:679–691.

    Article  PubMed  CAS  Google Scholar 

  46. Wonnacott, S., Wilkie, G., Soliakov, L., and Whiteaker, P. 1995. Presynaptic nicotinic autoreceptors and heteroreceptors in the CNS, in Effects Of Nicotine On Biological Systems II Ed. P. B. S. Clarke, M. Quik, F. Adlkofer & K. Thurau, Birkhauser Verlag, pp 87–94.

  47. Marley, P. D. 1988. Desensitisation of the nicotinic secretory response of adrenal chromaffin cells. Trends in Pharmacol. Sci. 9: 102–107.

    Article  CAS  Google Scholar 

  48. Grady, S., Marks, M., and Collins, A. C. 1994. Desensitisation of nicotine-stimulated [3H]dopamine release from mouse striatal synaptosomes. J. Neurochem. 62:1390–1398.

    Article  PubMed  CAS  Google Scholar 

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Special issue dedicated to Dr. Herman Bachelard.

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Wilkie, G.I., Hutson, P., Sullivan, J.P. et al. Pharmacological characterization of a nicotinic autoreceptor in rat hippocampal synaptosomes. Neurochem Res 21, 1141–1148 (1996). https://doi.org/10.1007/BF02532425

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