Larval zebrafish behavioral assays for wild-type (WT) and otofB mutants at 4 d postefertilization suggest altered behavior and GABAA receptor function in otofB mutants. (A, B) The larval photomotor response (LPR) assay evaluated locomotor activity in response to repeated photoperiod transitions from light (L) to dark (D), which typically elicits hyperactive responses in the dark phase. Five trials were conducted with 3 min per photoperiod. For the LPR assay, data are shown and statistically analyzed using a time-course plot of average locomotor for (A) each minute, as well as average activity within (B) each 3 min photoperiod. (C, D) The larval chemomotor response (LCR) assay evaluated locomotor activity in response to 100 µM picrotoxin (PTX), a known GABAA receptor antagonist that elicits a robust hyperactive locomotor response in larval zebrafish. Locomotor activity was measured for 3 h immediately following the application of PTX, relative to control (0.1% DMSO). For the LCR assay, average locomotor activity for each minute is plotted across the 3 h timecourse (C), and data were statistically analyzed using average activity within 10 min time bins (D). All data are reported as mean ± SEM. Significance was determined using two-way repeated measures ANOVA with Tukey’s post-hoc, p ≤ 0.05, N = 48 animals per group (4 d postfertilization larval zebrafish).
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