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Analytical Chem JoVE (Open Access) Citable · DOI

Patch Clamp Recordings in Inner Ear Hair Cells Isolated from Zebrafish

DOI: 10.3791/4281-v
What you'll learn
  • ✓ Isolate intact hair cells from zebrafish inner ear organs
  • ✓ Perform patch clamp electrophysiology on hair cell membranes
  • ✓ Characterize voltage-gated channel biophysical properties in sensory cells
Protocol

The purpose of this video is to demonstrate procedures for obtaining healthy, intact hair cells from the inner ear organs of adult zebrafish and then using them for patch clamp studies aimed at characterizing the biophysical properties of their voltage-gated channels.

Difficulty
advanced
Total time
~4–6 hours per zebrafish (isolation + recordings)
Model organism
Zebrafish (Danio rerio), adult
Biosafety
BSL-1

Steps

1
Prepare solutions and equipment for dissection

Set up recording chambers, pipettes, and physiological solutions required for hair cell isolation and patch clamp recording.

▶ 01:51
2
Extract auditory and vestibular labyrinth organs

Dissect the zebrafish inner ear to isolate the auditory and vestibular labyrinths under a microscope using fine surgical tools.

▶ 02:47
3
Enzymatically dissociate hair cells from tissue

Treat isolated labyrinth tissue with enzymatic and mechanical procedures to obtain individual, viable hair cells for electrophysiology.

▶ 05:28
4
Perform patch clamp recordings on hair cells

Use a patch clamp amplifier and micromanipulators to seal micropipettes onto hair cell membranes and record voltage-gated channel currents.

▶ 07:18
5
Analyze isolation success and electrophysiological data

Review microscopy images and electrophysiology traces to confirm healthy cell isolation and validate biophysical channel characterization.

▶ 10:12
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