Dissociating the effects of habituation, black walls, buspirone and ethanol on anxiety-like behavioral responses in shoaling zebrafish. A 3D approach to social behavior.

Understanding the different patterns of anxiety-like behavioral responses is of great interest for pharmacological and genetic research. Here we report the effects of 3.5-hr habituation, buspirone and ethanol on those responses in shoaling zebrafish (Danio rerio). Since in these experiments we used a container with white walls, the effects of black-vs.-white walls were tested in a separate experiment. An important objective was to determine whether factors unrelated to anxiety played a role in modulating the responses. The anxiety-like behavioral responses studied here are social cohesion, distance from bottom and bottom-dwell time, radial distribution (to study thigmotaxis), transparent-wall preference (to study escape responses), locomotion and freezing. The experimental conditions yielded distinctly different response patterns. Thigmotaxis was the most obvious response to white walls and it was significantly reduced after 3.5-hr habituation. It was not affected by any of the drugs. The reduction of social cohesion after 3.5-hr habituation and in the 0.5% ethanol group was probably the most interesting effect seen in this study. A role of anxiety herein was suggested but could not be established with certainty. Other hypotheses were also discussed. The large increase of distance-from-bottom resulting in swimming close to the water surface, which occurred in both buspirone groups and in the 0.5%-ethanol group, is most likely not an anxiolytic response, because of the discrepancy with the in the literature well-established time-course and the absence of any effect of 3.5-hr habituation or black walls on vertical measures. Finally, locomotion and duration freezing could not be specifically taken as indicators for the state of anxiety and the results concerning transparent-wall preference were not sufficient clear. We conclude that the neuronal and ethological mechanisms underlying the effects of habituation, white-aversion, buspirone and ethanol on anxiety-like behavioral responses are complex and need further exploration.