DISSOCIATED HIPPOCAMPAL CULTURES AND ACUTE CORTICAL SLICE PREPARATION

Rat hippocampal neurons were cultured from postnatal day 1–2 mixed sex rats as previously described (Bowen et al., 2017). Briefly, the hippocampus was dissected from postnatal day 1–2 rats and dissociated in papain. Neurons were seeded at a density of 150,000–200,000 cells on 18mm glass #1.5 coverslips coated with poly-D-lysine. Cells were maintained at 37°C, 5% CO2 for 14–16 days before fixation. For live imaging, dissociated hippocampal neurons were transfected with 400ng of Gephyrin-FingR DNA at 14–15 DIV, using Lipofectamine2000 according to the manufacturer’s protocol. To generate acute slices, 4–5 week old mice were perfused as described in (Hedrick and Waters, 2010) and 30 μm slices were cut from cortex.