GAD67+/− mice were generated as previously described (10). Exon 2 (the first coding exon) of the Gad1 gene was flanked by loxP sites using gene targeting in embryonic stem cells. Using FLP recombinase, the Sv-NeoR selectable gene was removed to produce Gad1lox/+ mice, which were interbred to generate phenotypically normal Gad1lox/lox mice. Gad1lox/lox mice were bred with Mox2-Cre mice to delete exon 2 in the germ-line (Gad1Δ/+ [GAD67+/−]) mice. Interbreeding of Gad1Δ/+ mice generated some mice that did not survive beyond the perinatal period, consistent with previously described Gad1-null mice (11). Age, sex, and litter-matched (10 males and 4 females) GAD67+/− and WT mice (n=7 per group) were euthanized at eight weeks of age and brains were removed, frozen, and stored at −80°C.
