Home › Analytical Chem › Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog (Rana catesbeiana)
Analytical Chem JoVE (Open Access) Citable · DOI

Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog (Rana catesbeiana)

DOI: 10.3791/55380-v
What you'll learn
  • ✓ Dissect and isolate the sacculus from bullfrog inner ear tissue
  • ✓ Prepare hair cells for biophysical measurement using enzymatic digestion
  • ✓ Measure hair bundle mechanical properties and spontaneous oscillation
Protocol

Biopharma Insights The American bullfrog's (Rana catesbeiana) sacculus permits direct examination of hair-cell physiology. Here the dissection and preparation of the bullfrog's sacculus for biophysical studies is described. We show representative experiments from these hair cells, including the calculation of a bundle's force-displacement relation and measurement of its unforced motion.

Difficulty
advanced
Total time
~3–4 hours per sacculus preparation
Model organism
American bullfrog (Rana catesbeiana)
Biosafety
BSL-1

Steps

1
Extract inner ear organs from bullfrog

Perform surgical dissection of the American bullfrog to remove the inner ear structures containing the sacculus. This establishes the tissue source for subsequent hair cell isolation.

▶ 00:58
2
Isolate sacculus from inner ear tissue

Carefully dissect and separate the sacculus from surrounding inner ear organs. This yields the sensory epithelium containing hair cells for physiological study.

▶ 03:52
3
Digest and mount sacculus in one-chamber preparation

Enzymatically treat the sacculus to isolate hair cells and mount the tissue in a single experimental chamber for biophysical measurement.

▶ 06:03
4
Digest and mount sacculus in two-chamber preparation

Prepare an isolated sacculus using enzymatic digestion and mount it across two separate chambers to enable independent manipulation and measurement of hair bundle mechanics.

▶ 06:51
5
Measure bundle force-displacement and oscillation

Record the mechanical response of hair bundles to applied displacement and quantify their spontaneous oscillatory motion using biophysical instrumentation.

▶ 10:08
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