Effects of lorazepam tolerance and withdrawal on GABAA receptor-operated chloride channels.

Mice were treated with 4 mg/kg of lorazepam for 7 days via implanted osmotic mini pumps. After chronic drug treatment, brains were assayed for GABA-mediated chloride flux (GABA-Cl-). Compared to control, brain membranes from lorazepam-tolerant mice were resistant to flunitrazepam stimulation of GABA-Cl-. Lorazepam tolerance did not affect [3H]diazepam binding affinity but did lower binding number slightly. Membranes from lorazepam-tolerant mice were cross-tolerant to both ethanol and phenobarbital stimulation of GABA-Cl-. Pentobarbital-stimulation of GABA-Cl- was equivalent in the two treatment groups. An increase in maximum inhibition of chloride flux produced by the benzodiazepine partial inverse agonist, n-methyl-beta-carboline-3-carboxamide (FG-7142) in membranes from lorazepam-tolerant mice was observed. FG-7142 was also found to be a more potent inhibitor of [3H]diazepam binding in membranes from lorazepam-tolerant mice. Withdrawal from chronic treatment by an acute injection with the benzodiazepine antagonist RO-15-1788 (flumazenil), restored functioning of the channel complex to control levels. There were no differences between membranes from control and lorazepam withdrawn mice in stimulation by flunitrazepam, ethanol, phenobarbital and pentobarbital or inhibition by FG-7142 of GABA-Cl-. [3H]Diazepam-saturated binding parameters and inhibition of binding by FG-7142 were similar. Chronic administration of lorazepam reduces the coupling between the benzodiazepine agonist site and the chloride channel and concomitantly increases coupling between the channel and the inverse agonist site. Furthermore, these findings offer neurochemical evidence for cross-tolerance to ethanol and phenobarbital after induction of lorazepam tolerance.