title: Weakening of ion-channel interactions of Na+ and Li+ in acetylcholine-receptor channels of frog skeletal muscle with an increase in agonist concentration.
abstract: The possibility that increases in agonist concentration beyond threshold levels may force changes in the character of high-conductance open states of skeletal muscle nicotinic acetylcholine receptor channels (nAChR) was examined by seeing whether differences in several critical ionic properties of nAChR currents could be detected with changes in agonist level. Single- and bi-ionic whole-cell currents of Na+ and Li+ in voltage-clamped frog (Rana pipiens) muscle fibers were measured during local superfusion of endplates with carbamylcholine (carb) at concentrations of 54 microm (low-carb) and 270 microM (high-carb). Three ionic properties that would be affected by changes in the open-state configuration of channel subunits were tested. First, ion-saturation characteristics. Peak Na+ and Li+ currents in low-carb trials showed sublinear dependence on ion concentrations from 0 to 60 mM with Km values of 78 (Na+) and 49 (Li+) mM and a power function slope of 0. 75 on double-log plot. In contrast, the concentration dependence of Na+ and Li+ currents in high-carb tests was linear through the origin with a power function slope of 1.02. Second, Na+/Li+ selectivity. The ratio of peak Na+ and Li+ currents in low-carb tests varied from 1.86 to 2.28 for ion concentrations of from 20 to 60 mM [mean = 2.02 +/- 0.06 (SEM)] whereas the ratio for high-carb trials ranged from only 1.29 to 1.52 [mean = 1.42 +/- 0.40 (SEM)]. Third, competitive interactions of Na+ and Li+ currents. Equimolar mixtures of Na+ and Li+ in low-carb tests produced bi-ionic inward currents which were never larger than the single-ion Na+ current alone, but bi-ionic currents at the high-carb level were always greater than the single-ion Na+ current, approximating the sum of the single-ion Na+ and Li+ currents in most cases. The results are consistent with a decrease in ion-channel binding at the high-carb level and support the possibility of agonist-induced changes in the high-conductance open-state configuration of nAChR subunits which result in a weakening of constraints on cation movements through the channel.
authors.affiliation: Department of Physiology, University of Tennessee, Memphis, 62 South Dunlap, Memphis, TN 38163, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Manthey
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002217:Carbachol / Q000494:pharmacology; D004553:Electric Conductivity; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D008094:Lithium / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D018482:Muscle, Skeletal / Q000502:physiology*; D018722:Nicotinic Agonists / Q000008:administration & dosage* / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D011894:Rana pipiens; D011978:Receptors, Nicotinic / Q000502:physiology*; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000502:physiology*
chemical_list: D007473:Ion Channels; D018722:Nicotinic Agonists; D011978:Receptors, Nicotinic; D015222:Sodium Channels; D002217:Carbachol; D008094:Lithium; D012964:Sodium; D008274:Magnesium