Standardizing the auditory evoked potential technique: Ground-truthing against behavioral conditioning in the goldfish carassius auratus

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 and tank composition has on AEPs. In the first experiment, the effect of testing tank composition and electrode placement on AEPs was determined by recording AEPs in the same individual fish in a steel and PVC cylindrical testing tank, and simultaneously recording AEPs from four locations and two different depths on each of 12 goldfish, Carassius auratus. Results from these studies show that tank composition has an effect AEP strength and hearing thresholds, with steel producing lower thresholds for all frequencies. Electrode placement and depth showed no significant effect on hearing thresholds.In the second experiment, the hearing sensitivity of 12 goldfish was measured using both classical conditioning and AEPs in the same setup. For behavioral conditioning, the fish were trained to reduce their respiration rate in response to a 5s sound paired with a brief shock. Once the behavioral audiogram was completed, the AEP measurements were made without moving the fish. The same sound stimuli were presented and the resultant evoked potentials were recorded for 1,000-6,000 averages. AEP input-output functions were then compared to the behavioral audiogram to compare techniques for estimating behavioral thresholds from AEP data. Results show a large range in variability between behavioral and evoked potential thresholds between fish, with the linear regression evoked potential analysis method producing closer thresholds to behavioral methods. In the third study, the effects of masking were examined on the behavioral and evoked potential audiograms. Behavioral thresholds were first determined with a constant masking noise for two frequencies, followed by threshold measurements with no masking noise