Data_Sheet_1_The effects of exploratory behavior on physical activity in a common animal model of human disease, zebrafish (Danio rerio).docx

Zebrafish (Danio rerio) are widely accepted as a multidisciplinary vertebrate model for neurobehavioral and clinical studies, and more recently have become established as a model for exercise physiology and behavior. Individual differences in activity level (e.g., exploration) have been characterized in zebrafish, however, how different levels of exploration correspond to differences in motivation to engage in swimming behavior has not yet been explored. We screened individual zebrafish in two tests of exploration: the open field and novel tank diving tests. The fish were then exposed to a tank in which they could choose to enter a compartment with a flow of water (as a means of testing voluntary motivation to exercise). After a 2-day habituation period, behavioral observations were conducted. We used correlative analyses to investigate the robustness of the different exploration tests. Due to the complexity of dependent behavioral variables, we used machine learning to determine the personality variables that were best at predicting swimming behavior. Our results show that contrary to our predictions, the correlation between novel tank diving test variables and open field test variables was relatively weak. Novel tank diving variables were more correlated with themselves than open field variables were to each other. Males exhibited stronger relationships between behavioral variables than did females. In terms of swimming behavior, fish that spent more time in the swimming zone spent more time actively swimming, however, swimming behavior was inconsistent across the time of the study. All relationships between swimming variables and exploration tests were relatively weak, though novel tank diving test variables had stronger correlations. Machine learning showed that three novel tank diving variables (entries top/bottom, movement rate, average top entry duration) and one open field variable (proportion of time spent frozen) were the best predictors of swimming behavior, demonstrating that the novel tank diving test is a powerful tool to investigate exploration. Increased knowledge about how individual differences in exploration may play a role in swimming behavior in zebrafish is fundamental to their utility as a model of exercise physiology and behavior.</p