title: Neuronal nicotinic acetylcholine receptors are blocked by intracellular spermine in a voltage-dependent manner.
abstract: A common feature of neuronal nicotinic acetylcholine receptors (nAChRs) is that they conduct inward current at negative membrane potentials but little outward current at positive membrane potentials, a property referred to as inward rectification. Physiologically, inward rectification serves important functions, and the main goal of our study was to investigate the mechanisms underlying the rectification of these receptors. We examined recombinant alpha3beta4 and alpha4beta2 neuronal nAChR subtypes expressed in Xenopus oocytes and native nAChRs expressed on superior cervical ganglion (SCG) neurons. Whole-cell ACh-evoked currents recorded from these receptors exhibited strong inward rectification. In contrast, we showed that single-channel currents from these neuronal nAChRs measured in outside-out patches outwardly rectify. On the basis of recent findings that spermine, a ubiquitous intracellular polyamine, confers rectification to glutamate receptors and inwardly rectifying potassium channels, we investigated whether spermine causes neuronal nAChRs to inwardly rectify. When spermine was added to the patch electrode in outside-out recordings, it caused a concentration- and voltage-dependent block of ACh-evoked single-channel currents. Using these single-channel data and physiological concentrations of intracellular spermine, we could account for the inward rectification of macroscopic whole-cell ACh-evoked conductance-voltage relationships. Therefore, we conclude that the voltage-dependent block by intracellular spermine underlies inward rectification of neuronal nAChRs. We also found that extracellular spermine blocks both alpha3beta4 and alpha4beta2 receptors; this finding points to a mechanism whereby increases in extracellular spermine, perhaps during pathological conditions, could selectively block these receptors.
authors.affiliation: Department of Physiology, McGill University, Montréal, Québec, Canada H3G 1Y6.
authors.forename: A P
authors.initials: AP
authors.lastname: Haghighi
authors.forename: E
authors.initials: E
authors.lastname: Cooper
mesh_terms: D000109:Acetylcholine / Q000493:pharmacokinetics; D000320:Adrenergic Fibers / Q000737:chemistry / Q000187:drug effects; D000818:Animals; D002478:Cells, Cultured; D004558:Electric Stimulation; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology*; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000737:chemistry / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000502:physiology*; D013096:Spermine / Q000494:pharmacology*; D017783:Superior Cervical Ganglion / Q000166:cytology; D014981:Xenopus
chemical_list: D011978:Receptors, Nicotinic; D013096:Spermine; D000109:Acetylcholine