Route choices during exploration of a complex maze shape were studied in humans and rats. For that purpose we designed a maze, the tree-maze, where subjects had to make two successive route choices. Different configurations of the maze were used in order to study the impact of the configuration in both species. A paper draw-through version of the task was also used with humans. Results of humans in the real version and in the paper draw-through version were very consistent. Humans and rats share a preference for following a straight path direction. Both species express a similar turning bias toward the right. The configuration had an impact on the choices of both species, but the general travelling directions, which was straight for humans and more to the right for rats, were different. Comparison between humans and rats remains difficult. Indeed, the stress induced by the situation was certainly different for both species. Nevertheless, the tree-maze seems to be a useful tool for studying route choices in different species.