Figure 3

Inhibition of TEA-induced GluR1 trafficking by surgery. Four-month-old Fischer 344 rats were subjected to right carotid artery exposure under isoflurane anesthesia with or without intravenous lidocaine. Hippocampus was harvested two weeks after surgery. Freshly prepared 300 μm coronal hippocampal slices were incubated with 25 mM TEA in the presence of 1 mg/ml sulfo-NHS-SS-biotin, a biotinylation reagent, for 10 minutes at 37°C. The homogenates of the hippocampal slices were used for western blotting of phospho-GluR1, phospho-PKA, and total GluR1 and the biotinylated fraction was used for western blotting of GluR1. A: biotinylated GluR1, B: phospho-GluR1 at ser845, C: total GluR1, and D: phospho-PKA. Representative western blot images are presented on the top of each panel and a graphic presentation of the results is shown in the bottom of each panel. Results are means ± SD (n = 8-11). *P <0.05 compared with the corresponding control. GAPDH, glyceraldehydes 3-phosphate dehydrogenase; P-GluR1, phospho-GluR1, PKA, protein kinase A; P-PKA, phospho-PKA; S-GluR1, cell surface GluR1; TEA, tetraethylammonium; T-GluR1: total GluR1.