Whole-Cell Patch Clamp Recording in a Substantia Gelatinosa Neuron of a Spinal Cord Slice
Transcript
Place a rat spinal cord slice in a recording chamber and secure it with a tissue anchor.
Perfuse the slice with artificial cerebrospinal fluid.
Under a microscope, identify the translucent substantia gelatinosa or SG region and select a healthy SG neuron.
Position a patch pipette filled with an ionic solution near the neuron.
Apply mild positive pressure to remove any debris from the pipette tip.
Move the pipette towards the neuron.
Then, release the positive pressure to form a dimple on the neuronal membrane, creating a seal.
Alter the holding potential to bring it close to the resting membrane potential, or RMP.
Use brief suction to rupture the membrane, establishing a whole-cell configuration.
At RMP, apply depolarizing electrical pulses.
These pulses open voltage-gated sodium channels, allowing a rapid influx of sodium ions.
This influx of ions triggers an action potential, resulting in neuron firing.
Record the neuron firing patterns.
