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.
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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
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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
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DOI: https://doi.org/10.1007/BF00370805