Effects of maternal separation in rats have been extensively investigated, but no studies have examined its effects in rat adolescence. We examined the effects of neonatal infant-mother separation (MS) for 6h/day and early handling (EH) for 10 days during the first 2 weeks of life by comparing MS and EH groups to standard facility reared (SFR) controls. At adolescence, the animals were evaluated in a novel and familiar open-field, the light-dark box, and the sucrose consumption test. Behavioral indices included orienting behavior (rearing frequency and duration), impulsive behavior (movement velocity and risk taking by entering the center of the open field or the light compartment of the light-dark box), hyperactivity (ambulatory distance and stereotypic movement), and reward-seeking behavior (sucrose drinking time). The prolonged MS during the first 2 weeks of life resulted in decreased orienting behavior and increased impulsive behavior in adolescence. Measures of ambulatory and stereotypic movements showed that MS rats were hyperactive in the novel environment whereas EH rats were less active overall. The impulsive/hyperactive phenotype produced by this MS protocol may provide a useful animal model to investigate the neurological basis for the similar behavioral phenotype found in attention deficit/hyperactivity disorder.