Using the Auditory Brainstem Response (ABR) to Determine Sensitivity of Hearing in Mutant Mice

Neil J. Ingham1, Selina Pearson1, Karen P. Steel1

1 Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo110059
Online Posting Date:  June, 2011
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Measurements of auditory evoked potentials can be used to determine reliably an audiometric representation of hearing sensitivity in mice. In a high‐throughput phenotyping screen of mice carrying targeted mutations of single genes, the auditory brainstem response (ABR) is used to gain an estimate of hearing threshold for broadband click stimuli and pure tone frequencies ranging from 6 to 30 kHz. Comparison of thresholds obtained in mutant and wild‐type mice give a means to determine mild, moderate, and severe hearing impairment. This gives a clear advantage over using a “clickbox” test to assess hearing by observations of the Preyer reflex. The ABR screen has identified several mutant lines with mild and moderate hearing loss, which appear to demonstrate normal Preyer responses. The ABR technique also allows frequency‐selective hearing loss to be identified. Curr. Protoc. Mouse Biol. 1:279‐287 © 2011 by John Wiley & Sons, Inc.

Keywords: hearing; evoked potential; electrophysiology; high‐throughput screening

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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
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Basic Protocol 1:

  • Male or female mice: control and mutant, tested at an appropriate age (no ABRs are likely to be recordable before the onset of hearing at around postnatal day 12)
  • Ketamine/xylazine anesthetic (see recipe)
  • Viscotears (liquid eye gel containing 2 mg/g Carbomer, with cetrimide; Dr Mann‐Pharma)
  • Atipamezole mix (recovery agent; see recipe)
  • Sound‐attenuating chamber (e.g., Industrial Acoustics)
  • Heating blanket (to prevent hypothermia in the mouse)
  • Stimulus‐generation and calibration equipment (Tucker‐Davis Technologies, TDT) including:
    • RP2.1 Enhanced Real‐time processor PA5 Programmable Attenuator
    • SA1 Stereo Amplifier
    • ACO Pacific Microphone (7017) and Preamplifier (PS9200) for calibration*
    • MA3 Stereo Microphone Amplifier
    • Sound Transducer (CTS Type 341; RS Components part no. 172‐7712)*
    • BNC and other connector cables
  • 1‐ml syringes (for injection; BD Plastipak)
  • 27‐G, 13‐mm length hypodermic needles (for injection; BD Microlance)
  • Response‐processing equipment (Tucker‐Davis Technologies) including:
    • Needle electrodes (Chalgren Enterprises, cat. no. 112‐812‐48‐TP; disposable low‐profile EEG needle electrodes; Fig. A)
    • Low‐Impedance Recording Headstage/Preamplifier (RA4LI + RA4PA)*
    • RA16 Medusa Base Station
    • RP2.1 Enhanced Realtime processor
    • HB7 Headphone Buffer (Optional)
    • MS2 Monitor Speaker (Optional)
    • BNC and other connector cables
  • Personal computer, housing TDT gigabit interface, TDT driver software and bespoke averager software (available on request)
  • Digital Oscilloscope (to view stimulus and electrode signals; optional)
NOTE: Items with an asterisk (*) next to them are housed within the sound attenuating chamber.
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Literature Cited

   Martin, G.K., Stagner, B.B., and Lonsbury‐Martin, B.L. 2006. Assessment of cochlear function in mice: Distortion product otoacoustic emissions. Curr. Protoc. Neurosci. 8.21C.1‐8.21C.18.
   Pauli‐Magnus, D., Hoch, G., Strenzke, N., Anderson, S., Jentsch, T.J., and Moser, T. 2007. Detection and differentiation of sensorineural hearing loss in mice using auditory‐steady state responses and transient auditory brainstem responses. Neuroscience 149:673–684.
   Spiden, S.L., Bortolozzi, M., Di Leva, F., Hrabe de Angelis, M., Fuchs, H., Lim, D., Ortolano, S., Ingham, N.J., Brini M., Carafoli, M., Mammano, F., and Steel, K.P. 2008. The novel mouse mutation Oblivion inactivates the PMCA2 pump and causes progressive hearing loss. PLoS Genetics 4:e1000238.
Internet Resources
  Web site for phenotyping and mutant mouse resources at the Wellcome Trust Sanger Institute.
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