Establish low-density, healthy dissociated primary hippocampal neuron cultures from E18 rat embryos that mature in vitro to form functional synapses (DIV14-21), providing a reproducible substrate for transfection, immunostaining, calcium imaging, and electrophysiology.
Quantify excitatory synapse density and structural integrity in dissociated hippocampal neurons by measuring colocalized synapsin-1 (presynaptic) and PSD-95 (postsynaptic) puncta along MAP2-defined dendrites.
Quantify the ratio of AMPA receptor-mediated to NMDA receptor-mediated excitatory postsynaptic currents (AMPA/NMDA ratio) at Schaffer collateral-CA1 synapses as a readout of postsynaptic synaptic strength and plasticity state in acute hippocampal slices.
Measure intracellular calcium dynamics in primary cortical neurons using the ratiometric indicator Fura-2 AM to quantify spontaneous network activity and depolarization-evoked calcium influx as readouts of neuronal excitability and network function.
Visualize complete dendritic arbors of cortical pyramidal neurons via Golgi-Cox impregnation and quantify dendritic complexity using Sholl analysis to detect treatment- or genotype-dependent changes in branching and arbor extent.