Ground-truthing evoked potential measurements against behavioral conditioning in the goldfish, Carassius auratus

Abstract

Auditory evoked potentials (AEPs) have become commonly used to measure hearing thresholds in fish. However, it is uncertain how well AEP thresholds match behavioral hearing thresholds and what effect variability in electrode placement has on AEPs. In the first experiment, the effect of electrode placement on AEPs was determined by simultaneously recording AEPs from four locations on each of 12 goldfish, Carassius auratus. In the second experiment, the hearing sensitivity of 12 goldfish was measured using both classical conditioning and AEP’s in the same setup. For behavioral conditioning, the fish were trained to reduce their respiration rate in response to a 5 s sound presentation paired with a brief shock. A modified staircase method was used in which 20 reversals were completed for each frequency, and threshold levels were determined by averaging the last 12 reversals. Once the behavioral audiogram was completed, the AEP measurements were made without moving the fish. The recording electrode was located subdermally over the medulla, and was inserted prior to classical conditioning to minimize handling of animal. The same sound stimuli (pulsed tones) were presented and the resultant evoked potentials were recorded for 1000–6000 averages. AEP input–output functions were then compared to the behavioral audiogram to compare techniques for estimating behavioral thresholds from AEP data.