Hippocampal neurons were dissected from newborn C57/BL6 wild-type mice and incubated in 0.25% trypsin-EDTA (Life Technologies) for 15 min at 37°C. After washing with Hank’s balanced salt solution plus 5 mM Hepes (Life Technologies), 20 mM d-glucose, and 2% fetal bovine serum (Gibco), the neurons were mechanically dissociated in culture medium and plated on poly-d-lysine–coated glass coverslips at a density of 50,000 to 100,000 cells/cm2. Cells were grown in Neurobasal-A medium (Life Technologies) supplemented with 2% B-27 (Life Technologies) and 2 mM GlutaMAX (Life Technologies). Cultures were maintained at 37°C in a 5% CO2-humidified incubator.

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