Abstract
Neuronal nicotinic acetylcholine receptors (nAChRs) were expressed in Xenopus laevis oocytes after nuclear injection of complementary deoxyribonucleic acid (cDNA) expression vectors. The two receptor subtypes α4/nα1 and α3/nα1 were readily distinguishable from one another by ACh sensitivity and desensitization. α3/nα1 receptors showed lower ACh sensitivity and stronger desensitization than α4/nα1 receptors. Furthermore, although the current/voltage relationship was very similar in both receptor subtypes, the voltage dependence of desensitization was found to be strikingly different. As the nα1 subunit was unchanged, the α subunits must be responsible for these functional differences. Symmetric hybrid α cDNAs, α4∶α3 and α3∶α4, were constructed and functional receptors were obtained by co-injection with nα1. These hybrid receptors displayed an ACh sensitivity that was mainly defined by the extracellular sequence of the α subunit. In contrast, no part of the α subunit was found fully to determine desensitization.
Similar content being viewed by others
References
Abramson SN, Taylor P (1990) Identification of nicotinic receptor ACh-binding subunits. Trends Neurosci 13:92
Ballivet M, Nef P, Couturier S, Rungger D, Bader CR, Bertrand D, Cooper E (1988) Electrophysiology of a chick neuronal nicotinic acetylcholine receptor expressed in Xenopus oocytes after cDNA injection. Neuron 1:847–852
Bertrand D, Bader CR (1986) DATAC: a multipurpose biological data analysis program based on a mathematical interpreter. Int J Bio-Medical Comput 18:193–202
Bertrand D, Ballivet M, Rungger D (1990) Activation and blocking of neuronal nicotinic acetylcholine receptor reconstituted in Xenopus oocytes. Proc Natl Acad Sci USA 87:1993–1997
Bertrand D, Cooper E, Valera S, Rungger D, Ballivet M (1991) Electrophysiology of neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes, following nuclear injection of genes or cDNAs. In: Conn PM (ed) Electrophysiology and microinjection, methods in neurosciences. Academic Press, New York, pp 174–193
Boulter J, Connolly J, Deneris E, Goldman D, Heinemann S, 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
Boyd RT, Jacob MH, Couturier S, Ballivet M, Berg DK (1988) Expression and regulation of neuronal acetylcholine receptor mRNA in chick ciliary ganglia. Neuron 1: 495–502
Cachelin AB, Colquhoun D (1989) Desensitization of the acetylcholine receptor of frog end-plates measured in a vaseline-gap voltage clamp. J Physiol (Lond) 415:159–188
Changeux JP, Devillers-Thiéry A, Chemouilli P (1984) Acetylcholine receptor: an allosteric protein. Science 225:1335–1345
Couturier S, Bertrand D, Matter JM, Hernandez MC, Bertrand S, Millar N, Valera S, Barkas T, Ballivet M (1990) A neuronal nicotinic acetylcholine receptor subunit (α7) is developmentally regulated and forms a homo-oligomeric channel blocked by α-Btx. Neuron 5:847–856
Couturier S, Erkman L, Valera S, Rungger D, Bertrand S, Boulter J, Ballivet M, Bertrand D (1990) α5, α3 and non-α3: three clustered avian genes encoding neuronal nicotinic acetylcholine receptor-related subunits. J Biol Chem 265:17560–17567
Deneris ES, Connolly J, Rogers SW, Duvoisin R (1991) Pharmacological and functional diversity of neuronal nicotinic acetylcholine receptors. Trends Pharmacol Sci 12:34–40
Dunn SMJ, Raftery MA (1982) Activation and desensitization of Torpedo acetylcholine receptor:evidence for separate binding sites. Proc Natl Acad Sci USA 79:6756–6761
Gross A, Bertrand D, Ballivet M, Rungger D (1990) Desensitization of a chicken neuronal nicotinic acetylcholine receptor expressed in Xenopus oocytes (abstract). Biophys J 57:119a
Hopfield JF, Tank DW, Greengard P, Huganir RL (1988) Functional modulation of the nicotinic acetylcholine receptor by tyrosine phosphorylation. Nature 336:677–680
Huganir RL, Delcour AH, Greengard P, Hess GP (1984) Phosphorylation of the nicotinic acetylcholine receptor regulates its rate of desensitization. Nature 321:774–776
Karlin A, Kao PN, DiPaola M (1986) Molecular pharmacology of the nicotinic acetylcholine receptor. Trends Pharmacol Sci 7:304–308
Kao PN, Karlin A (1986) Acetylcholine receptor binding site contains a disulfide crosslink between adjacent half-cystinyl residues. J Biol Chem 261:8085–8088
Kao PN, Dwork AJ, Kaldany RRJ, Silver ML, Wideman J, Stein S, Karlin A (1984) Identification of an α subunit half-cystine specifically labeled by affinity reagent for the acetylcholine receptor binding site. J Biol Chem 259:11662–11665
Mathie A, Colquhoun D, Cull-Candy SG (1990) Rectification of currents activated by nicotinic acetylcholine receptors in rat sympathetic ganglion neurons. J Physiol (Lond) 427:625–655
Nef P, Oneyser C, Alliod C, Couturier S, Ballivet M (1988) Genes expressed in the brain define three distinct neuronal nicotinic acetylcholine receptors. EMBO J 7:595–601
Ochoa ELM, Chattopadhyay A, McNamee MG (1989) Desensitization of the nicotinic acetylcholine receptor: molecular mechanisms and effect of modulators. Cell Mol Neurobiol 9:141–178
Sumikawa K, Miledi R (1989) Change in desensitization of cat muscle acetylcholine receptor caused by coexpression of Torpedo acetylcholine receptor subunits in Xenopus oocytes. Proc Natl Acad Sci USA 86:367–371
Unwin N, Toyoshima C, Kubalek E (1988) Arrangement of the acetylcholine receptor subunits in the resting and desensitized states, determined by cryoelectron microscopy of crystallized Torpedo postsynaptic membranes. J Cell Biol 107:1123–1138
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gross, A., Ballivet, M., Rungger, D. et al. Neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes: role of the α subunit in agonist sensitivity and desensitization. Pflügers Archiv 419, 545–551 (1991). https://doi.org/10.1007/BF00370805
Issue Date:
DOI: https://doi.org/10.1007/BF00370805