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Fig. 1 A battery of visual and acoustic assays elicits stereotyped behaviors in larval zebrafish. (a) Motor activity (y-axis) of zebrafish larvae exposed to 0.4 % DMSO for 40-min across various stimulus modalities (x-axis). Data are median ± 95 % CI for n = 5159 larvae obtained from 326 × 96-well plates each containing 24 control individuals. (b) A rapid transition from light to dark triggered a visual startle response during the baseline phase (VSRB). (c) Motor activity peaked approximately 5-min after the light-to-dark transition (BSL1), then gradually declined to baseline levels (BSL2, BSL3, BSL4). (d) A rapid dark-to-light transition caused a visual startle response (VSR1). (e) Following exposure to visual light, zebrafish larvae exhibit inactivity (VMR1). (f) Light-to-dark transition caused an additional visual startle response (VSR2). (g) Motor activity during the transition from light to dark reaches its peak around 5-min (VMR2), followed by an asymptotic decline to baseline levels (VMR3, VMR4, and VMR5). (h) Averaged spike from a series of five low- (ASR1) and (j,l) five high-intensity acoustic stimuli (1 s), provoking acoustic startle responses with low (ASR1) and high (ASR2, ASR3) amplitude, respectively. (i, k, m) Vehicle control were inactive during ISIs (ISIs) were calculated (ISI1, ISI2 and ISI3). (n) Responsiveness to non-habituating high-intensity acoustic stimuli significantly diminishes when comparing pre- (ASR2) and post-habituation (ASR3) assays, indicating the retention of memory. (o,p,q) Inter-acoustic endpoint intervals (IEI1, IEI2, IEI3). (r) A series of 30 high-intensity acoustic stimuli (interstimulus interval, ISI = 1 s) elicits an exponential decrement in response, indicative of habituation. (s) Representative plot of data used for r, excluding ISIs. (t) Aggregate sum of activity during acoustic stimulations for five habituation bouts (ASHsum). (u) Inter-habituation bout intervals (IBI). (v) Diminished responsiveness from the first (ASH1) relative to the fifth habituation bout (ASH5) indicates potentiation of habituation. Numbers above the split rainclouds in (n) and (v) represent p-values from a one-sided Wilcoxon signed-rank exact test. ASH, acoustic startle habituation; ASR, acoustic startle response; BSL, baseline motor activity; IBI, inter-bout interval; IEI, inter-endpoints interval; ISI, inter-stimulus interval; VMR, visual motor response; VSR, visual startle response; VSRB, visual startle response baseline.