In order to investigate the effects of early protein malnutrition and environmental stimulation upon the response to the anxiolytic properties of diazepam, two animal models of anxiety (elevated plus-maze and light-dark transition tests) were used. Rats were malnourished by feeding their dams a 6% protein diet during the lactation period (0-21 days of age) while well-nourished controls received a 16% protein diet. From 21 to 70 days of age all rats received a balanced lab chow diet. Environmental stimulation consisted of 3-min daily handling from birth to 70 days of age. Additional stimulation was provided from 21 to 70 days of age by rearing the rats in an enriched living cage. Eight groups of rats were studied in a 2 (malnourished or well-nourished) x 2 (stimulated or nonstimulated) x 2 (diazepam or vehicle) design. At 70 days of age, independent groups of rats treated with diazepam (2.5 mg/kg, IP) or vehicle were submitted to testing in the elevated plus-maze or light-dark transition procedures. The results showed that both diazepam and environmental stimulation reduced anxiety in the elevated plus-maze; stimulation changed the anxiolytic response to diazepam and the two diet conditions altered differentially the response to both pharmacological and stimulation procedures. These results suggest that environmental stimulation can affect differentially the behavioral response of malnourished and well-nourished rats treated with diazepam.