Patch-Clamp Recordings from the Dendrite of a Dopaminergic Neuron in a Brain Slice
Transcription
Begin with an electrophysiology setup with an immobilized brain slice submerged in an artificial cerebrospinal fluid or aCSF containing different ions.
The setup includes a patch pipette filled with a potassium-rich solution connected to a recording electrode and an amplifier.
Apply positive air pressure to the pipette to prevent its clogging. Then, position it near a dendrite, the signal-receiving extension of a dopaminergic neuron.
The resulting membrane dimpling confirms proximity to the dendrite.
Release the positive pressure to create suction, forming a tight seal between the pipette and the dendritic membrane.
Record the initial ion movement as an action current at the baseline voltage.
Flow-in aCSF with sodium and calcium channel blockers to block the respective channels, preventing ion movement through them.
Apply a negative voltage to open a voltage-gated cation channel.
This channel allows potassium ions to enter the neuron, increasing the action current and suggesting stimulation of the dendrite in a dopaminergic neuron.
