Summary
1. Chronic administration of nicotine up-regulates mammalian neuronal nicotinic acetylcholine receptors (nAChRs). A key hypothesis that explains up-regulation assumes that nicotine induces desensitization of receptor function. This is correlated with behaviorally expressed tolerance to the drug.
2. The present experiments were conducted to: (a) obtain information on the nicotine-induced desensitization of neuronal nAChR function, a less understood phenomenon as compared to that of the muscle and electric fish receptor counterparts; (b) test the hypothesis that different receptor subunit combinations exhibit distinct desensitization patterns.
3.Xenopus laevis oocytes were injected with mRNAs encoding rat receptor subunitsα2,α3, orα4 in pairwise combination with theβ2 subunit. The responses to various concentrations of acetylcholine (ACh) or nicotine were analyzed by the two electrode voltage clamp technique.
4. Concentration-effect curves showed that nicotine was more potent than ACh for all the receptor subunit combinations tested. Only theα4β2 combination exhibited a depression of the maximum effect at concentrations higher than 20µM nicotine.
5. After a single nicotine pulse, receptor desensitization (calculated as a single exponential decay) was significantly slower forα4β2 than for eitherα3β2 orα2β2.
6. Concentrations of nicotine that attained a near maximum effect were applied, washed, and re-applied in four minute cycles. The responses were calculated as percentages of the current evoked by the initial application. Following 16 minutes of this protocol, theα4β2 combination showed a greater reduction of the original response as compared to theα2β2 andα3β2 subunit combinations. Taking points 5 and 6 together, these experiments suggest that theα4β2 receptor subtype desensitizes at a slower rate and remains longer in the desensitized state.
7. Becauseα4β2 is the main receptor subunit combination within the brain and is up-regulated by nicotine, our data may be important for understanding the molecular basis of tolerance to this drug.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alkondon, M., and Albuquerque, E. X. (1993). Diversity of nicotinic acetylcholine receptors in rat hippocampal neurons. 1. Pharmacological and functional evidence for distinct structural subtypes.J. Pharm. Exp. Ther. 265:1455–1473.
Ballivet, M., Nef, P., Couturier, S., Rungger, D., Bader, C. R., Bertrand, D., and Cooper, E. (1988). Electrophysiology of a chick neuronal nicotinic acetylcholine receptor.Neuron 1:847–852.
Bertrand, D., Ballivet, M., and Rungger, D. (1990). Activation and blocking of neuronal nicotinic acetylcholine receptor reconstituted inXenopus oocytes.Proc. Natl. Acad. Sci. USA 87:1993–1997.
Bertrand, D., Devillers, T. A., Revah, F., Galzi, J.-L., Hussy, N., Mulle, C., Bertrand, S., Ballivet, M., and Changeux, J.-P. (1992). Unconventional pharmacology of a neuronal nicotinic receptor mutated in the channel domain.Proc. Natl. Acad. Sci. USA 89:1261–1265.
Boulter, J., Connolly, J., Deneris, E., Goldman, D., Heinemann, S., and Patrick, J. (1987). Functional expression of two neuronal nicotinic acetylcholine receptors from cDNA clones identifies a gene family.Proc. Natl. Acad. Sci. USA 84:7763–7767.
Boulter, J., Holloman, M., O'Shea-Greenfield, A., Duvoisin, R. M., Connolly, J. G., Wada, E., Jensen, A., and Gardner, P. D. (1990).α3,α5, andβ4: Three members of the rat neuronal nicotinic acetyltholine receptor-related gene family form a gene cluster.J. Biol. Chem. 265:4472–4482.
Boyd, N. D. (1987). Two distinct kinetic phases of desensitization of acetylcholine receptors of clonal rat PC12 cells.J. Physiol. (Lond.) 389:45–67.
Cachelin, A. B., and Jaggi, R. (1991).β-subunits determine the time course of desensitization in ratα-3 neuronal nicotinic acetylcholine receptors.Pflugers Arch. 419:579–582.
Carter, A. A., and Oswald, R. E. (1993). Channel blocking properties of a series of nicotinic cholinergic agonists.Biophys. J. 65:840–851.
Changeux, J.-P. (1990). Functional architecture and dynamics of the nicotinic acetylcholine receptor: An allosteric ligand-gated ion channel. In Changeux, J.-P., Llinas, R. R., Purves, D., and Bloom, F. E. (eds.),Fidia Res. Found. Neurosci. Award Lect., Raven Press, New York, pp. 21–168.
Charnet, P., Labarca, C., Cohen, B. N., Davidson, N., Lester, H. A., and Pilar, G. (1992). Pharmacological and kinetic properties ofα4β2 neuronal nicotinic acetylcholine receptors expressed inXenopus oocytes.J. Physiol. (Lon). 450:375–394.
Corriveau, R. A., and Berg, D. K. (1993). Coexpression of multiple acetylcholine receptor genes in neurons: Quantification of transcripts during development.J. Neurosci. 13:2662–2671.
Couturier, S., Bertrand, D., Matter, J.-M., Hernandez, M.-C., Bertrand, S., Millar, N., Valera, S., Barkas, T., and Ballivet, M. (1990a). A neuronal nicotinic acetylcholine receptor subunit (α7) is developmentally regulated and forms a homo-oligomeric channel blocked byα-BTX.Neuron 5:847–856.
Couturier, S., Erkman, L., Valera, S., Rungger, D., Bertrand, S., Boulter, J., Ballivet, M., and Bertrand, D. (1990b).α5,α3, and non-α3. Three clustered avian genes encoding neuronal nicotinic acetylcholine receptor-related subunits.J. BIol. Chem. 265:17560–17567.
Duvoisin, R. M., Deneris, E. S., Patrick, J., and Heinemann, S. (1989). The functional diversity of the neuronal nicotinic acetylcholine receptors is increased by a novel subunit:β4.Neuron 3:487–496.
Elgoyhen, A. B., Johnson, D. S., Boutler, J., Vetter, D. E., and Heinemann, S. (1994).α9—An acetylcholine receptor with novel pharmacological properties expressed in rat cochlear hair cells.Cell 79:705–715.
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 alpha 4 and beta 2 subunits and is up-regulated by chronic nicotine treatment.Mol. Pharmacol. 41:31–37.
Grady, S. R., Marks, M. J., and Collins, A. C. (1994). Desensitization of nicotine-stimulated [3H]dopamine release from mouse striatal synaptosomes.J. Neurochem. 62:1390–1398.
Gross, A., Ballivet, M., Rungger, D., and Bertrand, D. (1991). Neuronal nicotinic acetylcholine receptors expressed inXenopus oocytes: Role of theα subunit in agonist sensitivity and desensitization.Pflugers Arch. 419:545–551.
Higgins, L. S., and Berg, D. K. (1988). A desensitized form of neuronal acetylcholine receptor detected by3H-nicotine binding on bovine adrenal chromaffin cells.J. Neurosci. 8:1436–1446.
Hill, J. A., Zoli, M., Bourgeois, J.-P., and Changeux, J.-P. (1993). Immunocytochemical localization of a neuronal nicotinic receptor: theβ2 subunit.J. Neurosci. 13:1551–1568.
Leonard, J. P., and Snutch, T. (1991). The expression of neurotransmitter receptors and ion channels inXenopus oocytes. In Chad, J., and Wheal, H. (eds.),Molecular Neurobiology: A Practical Approach, Oxford University Press, New York, pp. 2–25.
Lester, R. A., and Dani, J. A. (1994). Time-dependent changes in central nicotinic acetylcholine channel kinetics in excised patches.Neuropharmacology 33:27–34.
Leutje, C. W., and Patrick, J. (1991). Bothα- andβ-subunits contribute to the agonist sensitivity of neuronal nicotinic acetylcholine receptors.J. Neurosci. 11:837–845.
Lukas, R. J. (1993). Expression of ganglia-type nicotinic acetylcholine receptors and nicotinic ligand binding sites by cells of the IMR-32 human neuroblastoma clonal line.J. Pharmacol. Exp. Ther. 265:294–302.
Marks, M. J., Burchs, J. B., and Collins, A. C. (1983). Effects of chronic nicotine infusion on tolerance development and cholinergic receptors.J. Pharmacol. Exp. Ther. 226:806–816.
Marks, M. J., Grady, S. R., Yang, J. M., Lippiello, P. M., and Collins, A. C. (1994). Desensitization of nicotine-stimulated86Rb+ efflux from mouse brain synaptosomes.J. Neurochem. 63:2125–2135.
Morales, A., and Sumikawa, K. (1992). Desensitization of junctional and extrajunctional nicotinic ACh receptors expressed inXenopus oocytes.Mol. Brain Res. 16:323–329.
Naranjo, D., and Brehm, P. (1993). Modal shifts in acetycholine receptor channel gating confer subunit-dependent desensitization.Science 260:1811–1814.
Nelson, M. E., and Albuquerque, E. X. (1994). 9-Aminoacridines act at a site different from that for Mg2+ in blockade of the N-methyl-D-aspartate receptor channel.Mol. Pharmacol. 46:151–60.
Ochoa, E. L., Li, L., and McNamee, M. G. (1990). Desensitization of central cholinergic mechanisms and neuroadaptation to nicotine.Mol. Neurobiol. 4:251–287.
Ochoa, E. L. M. (1994). Nicotine-related brain disorders: The neurological basis of nicotine dependence.Cell. Mol. Neurobiol. 14:195–225.
Ochoa, E. L. M., and O'Shea, S. M. (1994). Concomitant protein phosphorylation and endogenous acetylcholine release induced by nicotine: Dependency on neuronal nicotinic receptors and desensitization.Cell. Mol. Neurobiol. 14:315–340.
Papke, R. L., Boutler, J., Patrick, J., and Heinemann, S. (1989). Single-channel currents of rat neuronal nicotinic acetylcholine receptors expressed inXenopus oocytes.Neuron 3:589–596.
Patrick, J., Sequela, P., Vernino, S., Amador, M., Luetje, C., and Dani, J. A. (1993). Functional diversity of neuronal nicotinic acetylcholine receptors.Prog. Brain Res. 98:113–120.
Robinson, D., and McGee, R. (1985). Agonist-induced regulation of the neuronal nicotinic acetylcholine receptor of PC12 cells.Mol. Pharmacol. 27:409–417.
Rowell, P. P., and Hillebrand, J. A. (1994). Characterization of nicotine-induced desensitization of evoked dopamine release from rat striatal synaptosomes.J. Neurochem. 63:561–569.
Sands, S. B., and Barish, M. E. (1992). Neuronal nicotinic acetylcholine receptor currents in phaeochromocytoma (PC12) cells: Dual mechanisms of rectification.J. Physiol. (Lond.) 447:467–487.
Sargent, P. B. (1993). The diversity of neuronal nicotinic acetylcholine receptors.Annu. Rev. Neurosci. 16:403–443.
Schiavone, M. T., and Kirpekar, S. M. (1982). Inactivation of secretory responses to potassium and nicotine in the cat adrenal medulla.J. Pharmacol. Exp. Ther. 223:743–749.
Schwartz, R. D., and Kellar, K. J. (1985).In vivo regulation of [3H]acetylcholine recognition sites in brain by nicotinic cholinergic drugs.J. Neurochem. 45:427–433.
Seguela, P., Wadiche, J., Dineley, M. K., Dani, J. A., and Patrick, J. W. (1993). Molecular cloning, functional properties, and distribution of rat brainα7: A nicotinic cation channel highly permeable to calcium.J. Neurosci. 13:596–604.
Tandon, T., and Ochoa, E. L. M. (1992). Calcium and nicotine-induced desensitization of endogenous acetylcholine release form mammalian brain cholinergic nerve endings.Soc. Neurosci. Abstr. 18:634.
Vibat, C. R. T., Lasalde, J. A., McNamee, M. G., and Ochoa, E. L. M. (1994). Nicotine-induced desensitization of rat neuronal nicotinic acetylcholine receptor subunit combinations expressed inXenopus laevis oocytes.Soc. Neurosci. Abstr. 20:1128.
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 hybridization histochemical study in the rat.J. Comp. Neurol. 284:314–335.
Wada, K., Ballivet, M., Boulter, J., Connolly, J., Wada, E., Deneris, E. S., Swanson, L. W., Heinemann, S., and Patrick, J. (1988). Functional expressiom of a new pharmacological subtype of brain nicotinic acetylcholine receptor.Science 240.
Wallace, R. A., Jared, D. W., Dumont, J. N., and Sega, M. W. (1973). Protein incorporation by isolated amphibian oocytes: 3. Optimum incubation conditions.J. Exp. Zool. 184:321–333.
Wilkie, G. I., Hutson, P. H., Stephens, M. W., Whiting, P., and Wonnacott, S. (1993). Hippocampal nicotinic autoreceptors modulate acetylcholine release.Biochem. Soc. Trans. 21:429–431.
Wonnacott, S. (1991). Neuronal nicotinic receptors—functional correlates of ligand binding sites.Biochem. Soc. Trans. 19:121–124.
Wonnacott, S., Irons, J., Rapier, C., Thorne, B., and Lunt, G. G. (1989). Presynaptic modulation of transmitter release by nicotinic receptors.Prog. Brain Res. 79:157–163.
Yu, Z. J., and Wecker, L. (1994). Chronic nicotine administration differentially affects neurotransmitter release from rat striatal slices.J. Neurochem. 63:186–194.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vibat, C.R.T., Lasalde, J.A., McNamee, M.G. et al. Differential desensitization properties of rat neuronal nicotinic acetylcholine receptor subunit combinations expressed inXenopus laevis oocytes. Cell Mol Neurobiol 15, 411–425 (1995). https://doi.org/10.1007/BF02071877
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02071877