Recording Whole-Cell Currents with GABA Puffing in a Mouse Brain Slice
Transkript
Begin with a secured prefrontal cortical brain slice in an artificial cerebrospinal fluid, or aCSF, in the recording chamber of an electrophysiology setup.
This aCSF contains a sodium ion channel blocker, which inhibits sodium channels in neurons.
Position a puff micropipette filled with gamma-aminobutyric acid, or GABA, above a postsynaptic neuron.
Place an ionic solution-filled recording micropipette with an electrode near the same neuron.
Apply weak suction to create a tight seal with a small patch of the neuronal membrane.
Then, apply strong suction to disrupt the patched membrane, enabling access to the neuron's interior.
Deliver the GABA, an inhibitory neurotransmitter, which binds with a ligand-gated chloride channel on postsynaptic neurons.
This opens the channel and allows chloride ions to enter the neurons, generating an inhibitory postsynaptic current.
At a constant voltage, record the postsynaptic current. An increase in inhibitory postsynaptic current reflects GABA's role in neuronal activity inhibition.
