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Neuroscience JoVE (Open Access) Citable · DOI

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

DOI: 10.3791/50551-v
What you'll learn
  • ✓ Perform intact brain-spinal cord dissection from zebrafish embryos
  • ✓ Execute whole-cell patch clamp recordings from Mauthner neurons
  • ✓ Measure excitatory, inhibitory synaptic currents and action potentials
  • ✓ Identify voltage-gated channel activity in developing reticulospinal cells
Protocol

Biopharma Insights We have developed an intact brain-spinal cord preparation to record and monitor electrical activity via patch clamp recording from the Mauthner neurons and other reticulospinal cells in zebrafish embryos. Thus, we are able to record excitatory and inhibitory synaptic currents, voltage-gated channel activity and action potentials from key neurons in a developing embryo.

Difficulty
advanced
Total time
~4–6 hours per embryo (dissection, setup, recording session)
Model organism
Zebrafish (Danio rerio) embryo
Biosafety
BSL-1

Steps

1
Prepare and dissect embryonic zebrafish brain-spinal cord

Extract intact brain-spinal cord preparation from zebrafish embryo. Mount tissue in recording chamber under microscope for visualization of Mauthner neurons and reticulospinal cells.

▶ 01:18
2
Perform whole-cell patch clamp recording setup

Position micropipette electrode on target neuron (Mauthner cell), achieve gigaseal, and rupture cell membrane to establish whole-cell configuration. Configure amplifier and data acquisition for voltage and current measurements.

▶ 03:35
3
Record synaptic currents and action potentials

Acquire electrophysiological data from Mauthner neurons in voltage-clamp (synaptic currents) and current-clamp (action potentials) modes. Monitor voltage-gated channel activity and synaptic responses.

▶ 05:43
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