Home › Neuroscience › Application of a NMDA Receptor Conductance in Rat Midbrain Dopaminergic Neurons Using the Dynamic Clamp Technique
Neuroscience JoVE (Open Access) Citable · DOI

Application of a NMDA Receptor Conductance in Rat Midbrain Dopaminergic Neurons Using the Dynamic Clamp Technique

DOI: 10.3791/2275-v
What you'll learn
  • ✓ Prepare rat midbrain slices and establish whole-cell patch-clamp recordings from dopaminergic neurons
  • ✓ Apply NMDA receptor conductance to neurons using dynamic clamp technique in real-time
  • ✓ Interpret and analyze representative electrophysiological recordings from dynamic clamp experiments
Protocol

In this video, we demonstrate how to apply a conductance into a dopaminergic neuron recorded in the whole cell configuration in rat brain slices. This technique is called the dynamic clamp.

Difficulty
advanced
Total time
~3–4 hours per experiment (slice preparation ~1 hr, recordings ~2–3 hrs)
Model organism
Rat (Sprague Dawley, midbrain dopaminergic neurons)
Biosafety
BSL-1

Steps

1
Prepare rat midbrain tissue slices

Dissect and slice rat midbrain tissue to obtain thin sections suitable for electrophysiological recording. Maintain tissue viability in oxygenated artificial cerebrospinal fluid (ACSF).

▶ 01:17
2
Establish whole-cell patch-clamp recording configuration

Position dopaminergic neurons under microscope and form whole-cell patch-clamp recordings using microelectrodes. Optimize seal resistance and establish stable baseline membrane potential.

▶ 01:44
3
Apply NMDA receptor conductance via dynamic clamp

Inject simulated NMDA receptor conductance into the recorded neuron using real-time dynamic clamp software and hardware. Monitor voltage and current responses to virtual synaptic inputs.

▶ 04:09
4
Acquire and analyze representative recordings

Collect electrophysiological data showing neuron responses to applied conductance. Review and interpret voltage traces and current traces for evidence of NMDA receptor-like effects.

▶ 05:37
💬 Comments coming soon