Electrophysiological Recordings of Reprogrammed Neurons in a Mouse Brain Slice
Transcription
Take an immobilized genetically engineered mouse coronal slice in a recording chamber perfused with oxygenated buffer.
The slice contains the striatal region, which has interneurons reprogrammed from oligodendrocyte precursor cells marked by an interneuron marker and fluorescent protein expression.
Assemble a recording pipette consisting of an electrode immersed in an intracellular solution.
Microscopically identify a fluorescent interneuron.
Maintain a positive pressure in the pipette and advance it toward the target interneuron, contacting the membrane.
Apply negative pressure to form a high-resistance seal.
Apply negative pressure pulses to break the membrane, establishing a whole-cell configuration.
Maintain the interneuron's membrane potential at a constant negative value.
Apply brief incremental currents, opening the ion channels and changing the membrane potential, triggering an action potential.
Eventually, the membrane returns to the resting potential.
Observe these changes in membrane potential in response to the current injection, indicating neuronal maturation and successful reprogramming.
