title: Loreclezole inhibition of recombinant alpha1beta1gamma2L GABA(A) receptor single channel currents.
abstract: Loreclezole had two different effects on GABA(A) receptor (GABAR) currents. When applied to GABARs that contained a beta2 or beta3 subunit subtype, but not a beta1 subtype, loreclezole potentiated the peak current evoked by sub-maximal concentrations of GABA. Loreclezole also increased the rate and degree of apparent desensitization of GABAR whole-cell currents, an effect that was independent of the beta subunit subtype, suggesting that potentiation and inhibition of GABAR current by loreclezole occurred through separate sites. We used patch-clamp recording from outside-out and inside-out patches from L929 fibroblasts transiently transfected with rat GABAR subunits to examine the properties of inhibition of alpha1beta1gamma2L single channel currents by loreclezole. Loreclezole decreased the mean open time of the channel by decreasing the average durations of the open states. Loreclezole also increased the occurrence of a closed component with an average duration near 20 ms. Inhibition by loreclezole was not voltage-dependent. Loreclezole was equally effective when applied to the intracellular side of the receptor, suggesting that its binding site was readily accessible from both sides of the membrane. Pre-application of loreclezole effectively inhibited the GABAR current in macropatches, indicating that binding did not require an open channel. These findings were consistent with a mechanism of allosteric modulation at a site formed by the membrane spanning regions of the receptor.
authors.affiliation: Department of Neurology, University of Michigan Medical Center, Ann Arbor 48104-1687, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Fisher
authors.forename: D J
authors.initials: DJ
authors.lastname: Hinkle
authors.forename: R L
authors.initials: RL
authors.lastname: Macdonald
mesh_terms: D000818:Animals; D000927:Anticonvulsants / Q000494:pharmacology; D001921:Brain / Q000502:physiology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000187:drug effects / Q000502:physiology; D004594:Electrophysiology; D018757:GABA Modulators / Q000494:pharmacology; D058787:GABA-A Receptor Antagonists; D006801:Humans; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology; D051381:Rats; D011963:Receptors, GABA-A / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins; D014230:Triazoles / Q000494:pharmacology*
chemical_list: D000927:Anticonvulsants; D018757:GABA Modulators; D058787:GABA-A Receptor Antagonists; D020033:Protein Isoforms; D011963:Receptors, GABA-A; D011994:Recombinant Proteins; D014230:Triazoles; C066440:loreclezole