Epc 10 dual patch clamp amplifier

Recording micropipettes were pulled from borosilicate capillary glass (type 7052, 1.65 mm outer diameter; 1.12 mm inner diameter; World Precision Instruments) using a Flaming/Brown P-97 puller (Sutter Instruments) to obtain pipettes with a resistance of 3-5 MΩ for whole-cell recordings, and 2-3 MΩ for targeted extracellular recordings when filled with the appropriate pipette solution. Recording pipettes were wrapped with Parafilm to reduce capacitive transients. Recordings were made with an EPC-10 dual-patch clamp amplifier and Patchmaster software (HEKA Elektronik) running on a Macintosh computer.

Slices were transferred to a recording chamber and perfused with oxygenated ACSF (3 mL/min) and heated by an in-line heater (Warner Instruments) to 30 ± 1°C. GnRH-GFP neurons were identified by brief illumination at 470 nm using an upright fluorescence microscope Olympus BX51W1. Recording pipettes were pulled from borosilicate glass (type 7052, 1.65 mm outer diameter and 1.12 mm inner diameter; World Precision Instruments, Inc) using a P-97 puller (Sutter Instruments) to obtain pipettes with a resistance of 2–3.5 MΩ. Recordings were performed with an EPC-10 dual-patch clamp amplifier and Patchmaster acquisition software (HEKA Elektronik). Recorded cells were mapped to a brain atlas (Paxinos and Franklin, 2001 ) to determine if cell location was related to response to treatment. No such correlation was observed in this study.