Hippocampal Neuron Culture and Transfection

Standard hippocampal neuron culture and transfection were performed as described in our previous study (Zhang et al., 2012). Briefly, hippocampi were extracted from 1-day-old SD rats. After hippocampi were cut into pieces, they were digested using 0.125% trypsin. Finally, rat hippocampal neurons were plated onto a poly-D-lysine coated glass coverslip at a density of 1 × 10⁴ cells/cm². When cells were cultured for 13 days in vitro (DIV 13), different constructs were transfected into neurons using the calcium-phosphate method. All experiments were performed after transfection for 48 h.