Determination of GAB A Levels by a [3H]Muscimol Radioreceptor Assay

Abstract: A [³H]muscimol radioreceptor assay was used to measure the levels of GAB A in mouse brain. The method is based on the competitive inhibition of [³H]muscimol binding to the GABA receptor by GABA extracted from tissue. The specificity and accuracy of the method was established by comparative measurements of GABA levels by gas chromatography. GABA levels obtained by radioreceptor assay (R) and gas chromatography (GC) in different areas of mouse brain were (in μmol/g tissue ± S.E.M.): cerebral cortex 1.41 ± 0.06 (R), 1.50 ± 0.03 (GC); corpus striatum 1.70 ± 0.05 (R), 1.66 ± 0.01 (GC); cerebellum 1.15 ± 0.04 (R), 1.11 ± 0.07 (GC); hippocampus 1.35 ± 0.04 (R), 1.43 ± 0.04 (GC). The sensitivity of the assay was 5 pmol of GABA, which is sufficient to measure GABA levels in brain. The technique described is simple and rapid and it can be used for unpurified tissue extracts.