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 alpha 4/n alpha 1 and alpha 3/n alpha 1 were readily distinguishable from one another by ACh sensitivity and desensitization. alpha 3/n alpha 1 receptors showed lower ACh sensitivity and stronger desensitization than alpha 4/n alpha 1 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 n alpha 1 subunit was unchanged, the alpha subunits must be responsible for these functional differences. Symmetric hybrid alpha cDNAs, alpha 4:alpha 3 and alpha 3:alpha 4, were constructed and functional receptors were obtained by co-injection with n alpha 1. These hybrid receptors displayed an ACh sensitivity that was mainly defined by the extracellular sequence of the alpha subunit. In contrast, no part of the alpha subunit was found fully to determine desensitization.