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Effect of diazepam on γ‐aminobutyric acid (GABA) content of mouse brain

Diazepam (7-chloro1,3-dihydro1 -methyl-5-phenyl-2H1,4-benzodiazepin-2-one), has been reported to have anticonvulsant and antiepileptic effects (Banziger, 1965 ; Eidelberg, Harry & Miller, 1965; Galambos, 1965; Gastaut, Naquet & others, 1965). Highly significant correlation of y-aminobutyric acid (GABA) content in brain and excitability was suggested by Woodbury & Esplin (1959). The GABA content in the brains of epileptics was shown to be low (Elliott, Roberts & Baxter, 1959) and its formation decreased in brains of idiopathic epileptics and of rabbits sensitized to convulsions (Kokudo, 1959; Kuroda; 1959). For these reasons, the effect of diazepam on the normal cerebral GABA levels in the hemisphere and in hemispheres in which the GABA levels were reduced by the administration of isoniazid, was studied. GABA was quantitatively determined using the chromatographic method of Saad (1970). Both cerebral hemispheres, including white matter, from the brains of 3 decapitated animals were quickly frozen (-4 to -6") and pooled for each analysis. The elution of GABA spots was as described by Sytinskii & Priyatkina (1966) and the colour was read at 512 nm. Diazepam (Valium 10, amp., Hoffmann-La Roche) was administered intraperitoneally in doses of 5 or 10 mg/kg to adult male mice, 20-25 g, in 4 groups of 9 animals for each dose. Each group was subdivided into 3 subgroups from which the cerebral hemispheres were pooled and the GABA content determined 1, 2, 3 or 4 h after injection of diazepam. The mean GABA content for each group was compared with that from cerebral hemispheres of 9 untreated male mice in pooled samples each from 3 animals. The difference was considered significant if by Student's t-test P <0.05. The GABA content of cerebral hemispheres of 9 mice was depleted by isoniazid (Sugawara, 1958), administered (50 mg/kg, i.p.) and the GABA content determined for each subgroup of 3 animals 1 h later; this corresponds to the maximal fall in cerebral hemisphere GABA content (Saad, El Masry & Scott, 1972). Two groups of 9 mice were given isoniazid (50 mg/kg, i.p.) and diazepam (5 or 10 mg/kg, i.p.) separately, 1 h before decapitation and then the procedure was as described above. The mean GABA content for each group was compared with that of untreated animals, and that of isoniazid-treated animals. The state of activity of mice was observed before decapitation. 22 pg/g wet tissue. The effect of diazepam on the normal cerebral GABA levels in the hemispheres is summarized in Table 1. The mean control GABA content in the cerebral hemispheres was 415