Rapid seizure-induced alterations in mTORC1 upstream regulators BDNF, phospho-Akt (Thr308) and phospho-ERK1/2 (Thr202/Tyr204).

<p>A. Quantification of BDNF protein levels at 1, 3, 12, 24, 48 h post-HS shows a bi-phasic upregulation in the hippocampus at 1 h (1.31±0.12 fold, n = 7, p<0.05) and 12 h (1.25±0.08 fold, n = 7, p<0.01), and a single increase at 1 h (1.27±0.06 fold, n = 10, p<0.01) in the neocortex. Of note, only the mature 14 kD BDNF band is quantified because of the relevance of this isoform for synaptic plasticity and epileptogenesis. B. Phospho-Akt (Thr308) is significantly increased at 1 h post-HS in the hippocampus (1.24±0.09 fold, n = 8, p<0.05), while in the cortex levels are upregulated at 3 h (1.3±0.11 fold, n = 7, p<0.05) and downregulated at 24 h after HS (0.7±0.04 fold, n = 7, p<0.01). C. Phospho-ERK1/2 expression is significantly higher at 3 h post-HS in the hippocampus (1.46±0.18 fold, n = 7), but not in the neocortex (1.37±0.22 fold, n = 11, p = 0.11). Averaged optical density normalized to actin are expressed relative to the mean control values (dotted line). No significant difference is observed in the total levels of phospho-Akt (Thr308) or phospho-ERK1/2 (Thr202/Tyr204) (p>0.05). Insets are the representative western blots for individual proteins. C: control; HS: hypoxic seizures. *: p<0.05, **: p<0.01. Error bars indicate S.E.M.