Electrophysiological Recordings of Reprogrammed Neurons in a Mouse Brain Slice

JoVE Encyclopedia of Experiments

Neurowetenschap

0 weergaven • 2:14 min • July 8th, 2025

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

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.

After transferring one tissue section to a recording chamber for electrophysiology, mount the glass pipette on the recording electrode, and lower it into the solution. Double-check the resistance of the electrode. Then, slowly approach the reprogrammed cell with the pipette, keeping a slight positive pressure in the electrode to avoid plugging the tip, and check that the cell is GFP-positive before patching.

When the cell is patched, maintain the cell in current clamp from minus 60 to minus 70 millivolts, and inject 500-millisecond currents from minus 20 to plus 90 picoamperes with 10-picoampere increments to induce action potentials. This is indicative of a neuronal maturation and successful reprogramming.

07:23

Whole-cell patch-clamp opnamen in Brain Slices

Gerelateerde video's

0 Bekeken

11:52

Hele-cel patch-klem opnames van geïsoleerde primaire epitheliale cellen uit de epididymis

Gerelateerde video's

0 Bekeken

09:23

Auditieve Brainstem Response en buitenste Haarcel geheel-cel Patch klem opname bij postnatale ratten

Gerelateerde video's

0 Bekeken

03:52

An Electrophysiological Study of Retinogeniculate and Corticogeniculate Synapses in Mouse Brain Slices

Gerelateerde video's

0 Bekeken

03:05

Photostimulation and Whole-Cell Patch Clamp Recordings of Neurons in Mouse Hippocampal Slices

Gerelateerde video's

0 Bekeken

03:18

Optogenetic Stimulation and Electrophysiological Recording of Synaptic Events in Rat Brain Slices

Gerelateerde video's

0 Bekeken

10:04

Lokale toepassing van geneesmiddelen aan nicotine acetylcholine receptor functie in de hersenen van muizen Slices Studie

Gerelateerde video's

0 Bekeken

09:39

Direct-current stimulatie en multi-elektrode Array Registratie van Inbeslagneming-achtige activiteit in de muizen Brain Slice Voorbereiding

Gerelateerde video's

0 Bekeken

10:53

Voorbereiding van Acute hersenen segmenten met behulp van een geoptimaliseerde N-Methyl-D-glucamine beschermende herstelmethode

Gerelateerde video's

0 Bekeken

10:35

Twee-foton Calcium Imaging in neuronale Dendrites in hersenen plakjes

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 15 augustus 2026