Open field test locomotion & place preference during the OF and NOA tests. Zebrafish locomotor and place preference responses during the OF of the EC-response curve testing. (a) Average distance moved by concentration group. (b) Average time spent immobile by concentration group. (c) Average time spent in the center zone by concentration group. (d) Average time spent in the middle zone by concentration group. (e) Average time spent in the thigmotaxis zone by concentration group. Error bars represent S.E.M.

Novel object approach test locomotion & place preference during the OF and NOA tests. Zebrafish locomotor and place preference responses during the NOA of the EC-response curve testing. (a) Average distance moved by concentration group. (b) Average time spent immobile by concentration group. (c) Average time spent in the center zone by concentration group. (d) Average time spent in the middle zone by concentration group. (e) Average time spent in the thigmotaxis zone by concentration group. Error bars represent S.E.M. *p < 0.05, **p < 0.01, ***p < 0.001.

Open field test to novel object approach test post-hoc multifactor analysis. Evaluation of main and interaction effects between locomotor variables of the EC-response curve testing. (a) Average distance moved by concentration group and test. (b) Average time spent immobile by concentration group and test. (c) Average time spent mobile by concentration group and test. (d) Average time spent highly mobile by concentration group and test. Error bars represent S.E.M. Lines between points represent trendlines between concentrations.

Second open field test locomotion and place preference during the open field test to open field test procedure. Zebrafish locomotor and place preference responses during the OF-OF testing. (a) Average distance moved by concentration group. (b) Average time spent immobile by concentration group. (c) Average time spent in the thigmotaxis zone by concentration group. (d) Average time spent in the middle zone by concentration group. (e) Average time spent in the center zone by concentration group. Error bars represent S.E.M.

Open field test locomotion & place preference during with EtOH challenge. Zebrafish locomotoion and place preference responses during the comparison of FG-7142 with or without the presence of EtOH. (a) Average distance moved by concentration group. (b) Average time spent immobile by concentration group. (c) Average time spent in the center zone by concentration group. (d) Average time spent in the middle zone by concentration group. (e) Average time spent in the thigmotaxis zone by concentration group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Novel object approach test locomotion & place preference with EtOH challenge. Zebrafish locomotoion and place preference responses during the comparison of FG-7142 with or without the presence of EtOH. (a) Average distance moved by concentration group. (b) Average time spent immobile by concentration group. (c) Average time spent in the center zone by concentration group. (d) Average time spent in the middle zone by concentration group. (e) Average time spent in the thigmotaxis zone by concentration group. **p < 0.01, ***p < 0.001, ****p < 0.0001.

Opercular movement testing. Zebrafish opercular BPM while in habitat water, habitat water and FG-7142, or habitat water and ethanol. (a) Average opercular BPM by concentration group. (b) Average opercular BPM by concentration group and time in 2-min intervals. Error bars represent S.E.M. Lines between points represent trendlines over time between concentration group mean BPMs.

Visualization of experimental setup and design. The illustrated materials and procedure of the EC-response curve testing as well as arena placement. (a) Procedure and design of the OF-NOA testing. (b) Top-down demonstration of OF-NOA arena placement, stimuli, and temperature reading. (c) Top-down demonstration of opercular movement testing.