This led us to hypothesize that metabolic control by BAD may also influence seizure sensitivity in vivo, similar to the KD ( Stafstrom and Rho, 2004). To test this hypothesis, we used the chemical proconvulsant kainic acid (KA), which induces acute seizures by stimulating excitatory glutamate receptors ( Ben-Ari et al., 1980). Published studies in mice indicate that KD can delay the onset of KA-induced seizures ( Jeong et al., 2011 and Noh et al., 2003). The seizure responses in BAD mutant and control mice treated with KA were scored using a modified Racine scale as previously described ( Ferraro et al., 1997). Wild-type mice experienced

an acute, yet transient, series of seizures that peaked on average between 50–120 min after KA administration and then slowly decayed Autophagy Compound Library ic50 (Figures 3A and 3B). The seizure diaries for individual mice are shown in Figure S2. The majority of wild-type mice (>80%) underwent status epilepticus CP 868596 with very severe tonic-clonic seizures. In striking contrast, Bad−/− mice did not progress in severity to the extent of wild-type animals and were significantly protected from status epilepticus ( Figure 3A). BadS155A mice were similarly resistant to behavioral seizures in this experimental model ( Figure 3B). In addition to raw seizure scores based on the modified Racine scale, we integrated individual

scores per mouse over the duration of the experiment to better account for seizure severity in mice that died during the experiment ( Figure 3C). Seizure severity was significantly diminished in both Bad null and S155A knockin mice ( Figure 3C). Moreover, seizure resistance in the absence of BAD is not limited to the kainate model. Bad null mice were also protected against status epilepticus isothipendyl triggered by pentylenetetrazole (PTZ), which antagonizes GABAergic inhibitory receptors ( Bough and Eagles, 1999 and Ferraro et al., 1999; Figure 3D). To determine whether seizure resistance in Bad−/− and BadS155A mice is accompanied

by neurobehavioral abnormalities, we conducted a detailed battery of cognitive and motor function tests. These studies did not reveal any significant differences in Bad genetic models compared with control mice ( Figure S3). It therefore appears unlikely that these models produce major circuit-level effects that impair normal brain function and might also disrupt seizure mechanisms. As the Bad null and S155A alleles have opposite effects on BAD’s apoptotic function ( Danial, 2008), the comparable resistance to seizures observed in Bad null and S155A mice cannot be explained by changes in apoptosis. To further test for the specificity and selectivity of BAD in this paradigm, BID-deficient mice were examined. BID (BH3-Interacting domain Death agonist) is another BH3-only proapoptotic member of the BCL-2 family similar to BAD but does not affect glucose metabolism ( Danial, 2008).