Figure 5
Juvenile TSC2+/− mice exhibit a reduced seizure threshold that is restored to WT-like latency with SAHA. (A) A schematic of the injection paradigm used in this study. Juvenile WT and TSC2+/− mice were intraperitoneally injected with either 50 mg/kg SAHA or vehicle (100 mM HPβCD) at the time of injection. Immediately following flurothyl induced seizures, a subset of mice was sacrificed, and whole hippocampal and cortical tissue was collected for post hoc western blot analysis. (B) Juvenile TSC2+/− mice exhibit a reduced seizure threshold in response to flurothyl compared to age matched WT mice (Student t-test: p = 0.0346). Compared to those treated with vehicle (HPβCD), SAHA increased latency to GTCS in juvenile TSC2+/− mice (Student t-test: p = 0.01) to levels that are indistinguishable from untreated WT mice. Compared to vehicle treated WT mice, WT mice treated with SAHA do not exhibit changes in latency to GTCS. (C) Following flurothyl induction, the hippocampi of vehicle or SAHA treated mice were rapidly harvested and processed for western blot analysis. Representative cropped western blot of whole hippocampal lysate extracted from vehicle or SAHA treated mice post flurothyl induction shows a global increase in acetylated histone H3 protein in SAHA treated mice, suggesting SAHA crossed the blood brain barrier and had a physiological effect. Each lane represents lysate from a single animal. An n = 3 per condition is represented in the blot. (D) Whole hippocampal extracts acquired from juvenile WT and TSC2+/− mice injected with either SAHA or vehicle were subject to western blot analysis. Quantification of protein levels shows that compared to vehicle treated mice, SAHA treatment increases pan acetyl histone H3 protein levels in the hippocampus of both WT mice (Student t-test:p = 0.03) and TSC2+/− mice (Student t-test: p = 0.027).
