Dissociated Hippocampal Neuron Culture

All animal procedures were approved by the University of Maryland Animal Use and Care committee. Dissociated hippocampal neuron cultures were prepared from both male and female E18 Sprague Dawley rat embryos (Charles River) as described previously (Dharmasri et al., 2023; Divakaruni et al., 2018 (link)). Neurons were plated on glass coverslips (18 mm #1.5, Warner Instruments) coated with poly-l-lysine at 30,000 cells per coverslip. Cells were grown in Neurobasal A + GlutaMax, gentamycin, and B27 supplement, with FUDR added at 1 week to suppress glial growth. Cells were fixed between 13–15 DIV for most experiments, between 6–8 or 20–22 DIV for developmental comparisons, and at 21 DIV for experiments with bassoon labeling.