High-throughput behavioral screening method for detecting auditory response defects in zebrafish.
We have developed an automated, high-throughput behavioral screening method for detecting hearing defects in zebrafish. Our assay monitors a rapid escape reflex in response to a loud sound. With this approach, 36 adult zebrafish, restrained in visually isolated compartments, can be simultaneously assessed for responsiveness to near-field 400 Hz sinusoidal tone bursts. Automated, objective determinations of responses are achieved with a computer program that obtains images at precise times relative to the acoustic stimulus. Images taken with a CCD video camera before and after stimulus presentation are subtracted to reveal a response to the sound. Up to 108 fish can be screened per hour. Over 6500 fish were tested to validate the reliability of the assay. We found that 1% of these animals displayed hearing deficits. The phenotypes of non-responders were further assessed with radiological analysis for defects in the gross morphology of the auditory system. Nearly all of those showed abnormalities in conductive elements of the auditory system: the swim bladder or Weberian ossicles.
Department of Otolaryngology, Harvard Medical School and The Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA 02114, USA.
Pascal I
PI
Bang
Pamela C
PC
Yelick
Jarema J
JJ
Malicki
William F
WF
Sewell
D000161:Acoustic Stimulation; D000400:Air Sacs / Q000503:physiopathology; D000818:Animals; D006309:Hearing / Q000502:physiology*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D009154:Mutation; D013216:Reflex, Startle / Q000502:physiology; D013057:Spectrum Analysis; D015027:Zebrafish