Animal experiments were approved specifically for this study by the Center for Laboratory Animal Research, Tohoku University (approval number: 2014BeA-005-1). Timed-pregnant Sprague-Dawley rats were obtained from Charles River Laboratories, Japan, and were used immediately after receipt. Primary neurons were obtained from the cortices of embryonic day 18 pups, plated on the microfabricated coverslip, and cocultured with astrocyte feeder cells in N2 medium [MEM + N2 supplement + ovalbumin (0.5 mg ml−1) + 10 mM Hepes] (13, 14). Half the medium was changed at 4 and 8 DIV with Neurobasal medium [Neurobasal (21103-049, Gibco) + 2% B-27 supplement (17504-044, Gibco) + 1% GlutaMAX-I (35050-061, Gibco)].

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