MOLECULAR MECHANISMS OF THE PARTIAL ALLOSTERIC MODULATORY EFFECTS OF BRETAZENIL AT GAMMA-AMINOBUTYRIC-ACID TYPE-A RECEPTOR

In central nervous system gamma-aminobutyric acid (GABA) inhibits neuronal activity by acting on GABA type A (GABA(A)) receptors. These heterooligomeric integral membrane proteins include a GABA-gated Cl- channel and various allosteric modulatory sites where endogenous modulators and anxiolytic drugs act to regulate GABA action. In vivo, various anxiolytic drugs exhibit a wide range of variability in their modulatory efficacy and potency of GABA action. For instance, bretazenil modulatory efficacy is much lower than that of diazepam. Such low efficacy could be due either to a preferential modulation of specific GABA(A) receptor subtypes or to a low modulatory efficacy at every GABA(A) receptor subtype. To address these questions we studied drug-induced modifications of GABA-activated Cl- currents in native GABA(A) receptors of cortical neurons in primary cultures and in recombinant GABA(A) receptors transiently expressed in transformed human embryonic kidney cells (293) after transfection with cDNAs encoding different molecular forms of alpha, beta, and gamma-subunits of GABA(A) receptors. In cortical neurons the efficacy of bretazenil was lower than that of diazepam, whereas the potency of the two drugs was similar. In cells transfected with gamma-2 subunits and various molecular forms of alpha and beta-subunits bretazenil efficacy was always lower than that of diazepam. However, in cells transfected with gamma-1 or gamma-3 subunits and various forms of alpha and beta-subunits the efficacy of both diazepam and bretazenil was lower and always of similar magnitude. When bretazenil and diazepam were applied together to GABA(A) receptors including a gamma-2 subunit, the action of diazepam was curtailed in a manner related to the dose of bretazenil.