Modulation of benzodiazepine agonist and inverse-agonist receptor binding by GABA during ethanol withdrawal.

1. The present study examined the capacity of GABA to modulate flunitrazepam and Ro15-4513 binding to putative GABAA receptors. Binding was measured in distinct brain regions both before and during selected periods of withdrawal from ethanol. 2. Rats were fed a nutritionally complete liquid ethanol (4.5% w/v) diet for 4 days and at various times after the last dose of ethanol (0, 12, 24, & 72 hr), rats were sacrificed and extensively washed brain membrane fractions were prepared. 3. Competitive inhibition of 3H-flunitrazepam binding by either flunitrazepam or Ro15-4513 (10(-10)M to 10(-7)M) was performed in the absence and presence of GABA (10(-5)M). In the presence of GABA, the apparent affinity for flunitrazepam was increased approximately 1.7 fold and the apparent affinity for Ro15-4513 was decreased by 1.7 fold. 4. No alteration in the capacity of GABA to modulate flunitrazepam or Ro15-4513 affinity (e.g. GABA-shift) was observed in cortical membrane preparations either 12 or 72 hr following ethanol cessation. 5. Further, no changes in GABA-modulation of flunitrazepam binding was evident 0, 12, 24, or 72 hr after the last ethanol dose in membranes prepared from cortex, hippocampus or cerebellum. 6. Therefore, results from the present study indicate that the capacity of GABA to modulate receptor affinity for benzodiazepine agonists and inverse-agonists in rat cortex, hippocampus or cerebellum is not altered during withdrawal from chronic ethanol.