Proliferating cells after NMDA treatment.

<p><b>A:</b> Representative confocal fluorescence images of EdU labeling (red) and cell-specific markers (green) in the pyramidal cell layer at 4 day after NMDA treatment. EdU (10 µM) was contained in the culture medium for 48 h during 2–4 days after NMDA treatment. After EdU detection, slice cultures were immunostained using the primary antibodies as follows: anti-Iba1 for microglia, anti-NeuN for neurons, anti-GFAP for astrocytes and anti-NG2 for NG2-positive cells. Scale bars = 50 µm. <b>B, C:</b> Iba1/EdU-double positive cells in the pyramidal cell layer (P) and stratum radiatum (R) of vehicle- or NMDA-treated slice cultures were counted. EdU was contained in the culture medium for 48 h during 0–2 days (<b>B</b>) or 2–4 days (<b>C</b>) after vehicle or NMDA treatment. Cell counting was carried out in the areas of 200 µm×200 µm at two different z-positions (6.9-µm intervals). C, vehicle; N, NMDA. Data are expressed as the mean ± SEM from twelve areas of six slices.