title: The acetylcholinesterase inhibitor BW284c51 is a potent blocker of Torpedo nicotinic AchRs incorporated into the Xenopus oocyte membrane.
abstract: This work was aimed to determine if 1,5-bis(4-allyldimethylammoniumphenyl)pentan-3-one dibromide (BW284c51), the most selective acetylcholinesterase inhibitor (AchEI), affects the nicotinic acetylcholine (Ach) receptor (AchR) function. Purified Torpedo nicotinic AchRs were injected into Xenopus laevis oocytes and BW284c51 effects on Ach- and carbamylcholine (Cch)-elicited currents were assessed using the voltage-clamp technique.BW284c51 (up to 1 mM) did not evoke any change in the oocyte membrane conductance. When BW284c51 (10 pM-100 microM) and Ach were co-applied, Ach-evoked currents (I(Ach)) were reversibly inhibited in a concentration-dependent manner (Hill coefficient, 1; IC(50), 0.2-0.5 muM for 0.1-1000 microM Ach). Cch-elicited currents showed a similar inhibition by BW284c51.I(Ach) blockade by BW284c51 showed a strong voltage dependence, being only apparent at hyperpolarising potentials. BW284c51 also enhanced I(Ach) desensitisation.BW284c51 changed the Ach concentration-dependence curve of Torpedo AchR response from two-site to single-site kinetics, without noticeably affecting the EC(50) value. The BW284c51 blocking effect was highly selective for nicotinic over muscarinic receptors. BW284c51 inhibition potency was stronger than that of tacrine, and similar to that of d-tubocurarine (d-TC). Coapplication of BW284c51 with either tacrine or d-TC revealed synergistic inhibitory effects. Our results indicate that BW284c51 antagonises nicotinic AchRs in a noncompetitive way by blocking the receptor channel, and possibly by other, yet unknown, mechanisms. Therefore, besides acting as a selective AchEI, BW284c51 constitutes a powerful and reversible blocker of nicotinic AchRs that might be used as a valuable tool for understanding their function.
authors.affiliation: Departamento de Fisiología, Genética y Microbiología, División de Fisiología, Universidad de Alicante, Campus San Vicente, Aptdo. 99, Alicante E-03080, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Olivera-Bravo
authors.forename: Isabel
authors.initials: I
authors.lastname: Ivorra
authors.forename: Andrés
authors.initials: A
authors.lastname: Morales
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000737:chemistry / Q000494:pharmacology*; D002217:Carbachol / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry; D018678:Cholinergic Agents / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004553:Electric Conductivity; D005260:Female; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D008845:Microinjections; D015394:Molecular Structure; D018377:Neurotransmitter Agents / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology*; D009865:Oocytes / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013619:Tacrine / Q000494:pharmacology; D014101:Torpedo; D014981:Xenopus
chemical_list: D018678:Cholinergic Agents; D002800:Cholinesterase Inhibitors; D008565:Membrane Proteins; D018377:Neurotransmitter Agents; D018733:Nicotinic Antagonists; D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; D013619:Tacrine; D002217:Carbachol; D000109:Acetylcholine