2.5. Two-electrode voltage clamp electrophysiology

Approximately 48 h after injection, electrophysiology was performed using the Axoclamp 900A voltage clamp. Responses were recorded using varying concentrations of GABA dissolved in non-supplemented ND96. Microelectrodes were made using Ag|AgCl wires and filled with 3M KCl. Injected X. laevis oocytes were clamped and held at −60 mV and left for 1–2 min to stabilize before GABA was washed over the oocytes at a flow rate of 8 mL/min. For dose-response analysis, oocytes were washed with ND96 for 1–2 min between agonist applications. Data was recorded and analyzed using Clampex 10.3 software and graphs were produced using Graphpad Prism Software 5.0. Data was collected from at least five individual oocytes. To assure consistency, two different batches of oocytes were used.