Acoustic Startle Response in Zebrafish Larvae

Acoustic startle was performed as described previously (40 (link)) with minor modifications. Experiments were performed between 1 and 6 p.m. Behavior was assessed at 5 dpf in the presence of the drug, or at 6 dpf, in the absence of the drug, 24 h after the end of the exposure. Larvae were individually placed in 96-well plates, pseudo-randomized by drug treatment, and allowed to acclimate for 30 min. After this period, the plate was moved to a DanioVision Observational Chamber (Noldus Information Technology, Wageningen, The Netherlands) where, following a 2-min acclimatization period, larval movement was recorded. Following an initial 1 min baseline period, larvae were subjected to 25 sound/vibration stimuli with 2 s inter-stimulus intervals. The distance traveled was recorded using Ethovision XT software (Noldus Information Technology, Wageningen, The Netherlands), and data were output in 1 s time bins.