We use the patch-clamp technique to measure GABA-activated single-channel currents (GABAA channels, GABAA receptors) and the synaptic and tonic currents they generate in neurons. Activation of the channels decreases neuronal excitability in health and disease 1,2,3,4.
Total time
~4–6 hours per experiment (slice preparation, pipette fabrication, recordings)
Model organism
Rat (brain tissue)
Steps
1
Prepare acute brain slices from rat brain
Dissect rat brain and slice using a vibratome to obtain thin sections containing target neurons. Maintain slices in oxygenated artificial cerebrospinal fluid (ACSF) at physiological temperature.
▶ 01:42
2
Fabricate micropipettes for patch-clamp recording
Pull glass capillaries to create fine-tipped electrodes with appropriate resistance (2–10 MΩ). Polish pipette tips to achieve optimal cell sealing.
▶ 03:17
3
Perform patch-clamp recording from neurons
Apply GABA to neurons and use voltage-clamp electrophysiology to record single-channel and whole-cell currents. Position micropipette, form gigaseal, and measure GABAA receptor-mediated inward currents.
▶ 04:00
4
Analyze representative single-channel and tonic currents
Display representative current traces showing GABA-activated channel openings, synaptic events, and tonic baseline currents. Quantify current amplitudes and kinetics to characterize neuronal inhibition.
▶ 08:07