Protocol to study chloride regulation in cultured mouse cortical neurons using electrophysiology

Inhibitory synaptic transmission mediated by the neurotransmitter γ-aminobutyric acid (GABA) is dependent on the concentration of chloride ions (Cl^-) in neurons, which can be assessed by making patch-clamp recordings of the reversal potential for GABA (E_GABA). Here, we present a protocol to study the regulation of cation-chloride cotransporters and the strength of synaptic inhibition in cultured mouse cortical neurons using electrophysiology. We describe steps for culturing neurons isolated from postnatal pups and electrophysiological measurement of E_GABA. For complete details on the use and execution of this protocol, please refer to Raveendran et al.¹.