PO 0.75 mg/day in 3 divided doses.

An aliquot (1 ml) of synaptosomal membrane suspension (1 mg protein) was added to the assay tubes containing FLUDIAZEPAM at varying concentrations (from 0.1 nM to 104 nM) and [3H]-diazepam (0.6 nM) in 2 ml of 50 mM Tris-HCl buffer (pH 7.4). The assay mixtures were incubated at 4 ° C for 15 min. The incubation was terminated by adding ice-cold 50 mM Tris-HCl buffer (5 ml) followed by rapid filtration through a Whatman GF/B filter. Each filter was immediately washed twice with ice-cold 50 mM Tris-HCl buffer (5 ml) The bound [3H]-diazepam retained on the filter was extracted with ACS II (Amersham)(10 ml) and counted. All incubations were done in triplicate. Nonspecific binding was determined in tubes containing diazepam at a final concentration of 103 nM.