Prenatal restraint stress (PS) has been suggested as an attractive chronic model of anxiety. Here, we characterized the behavioral and autonomic responsivity to acute stress exposure in adult PS subjects. In Wistar rats, locomotor activity, as well as spontaneous behavior in an established animal model of anxiety, the elevated plus-maze, was unaffected by PS. However, the anxiogenic-like response normally seen on the plus-maze following a restraint stress was markedly potentiated in adult PS subjects, despite indistinguishable corticosterone responses. In addition, we assessed the tail skin temperature response to a mild social stressor, transient social mixing. The diazepam-sensitive, late phase of the temperature response was markedly potentiated in adult PS subjects. In summary, PS induces a persistent phenotype of increased behavioral and autonomic sensitivity to stress. This paradigm might serve as an attractive screening model for anti-anxiety compounds.