We investigated prolactin secretion and metabolic changes in stress response in adult male rats submitted to periodic maternal separation (MS; 180 min/day) at 2 weeks of life. Restraint and ether exposure were randomly performed when the animals were 10–12 weeks of age. Restraint exposure: the animals were placed into plastic tubes (21 cm long, 4.5 cm diameter) for 20 min. Ether exposure: the rats were exposed to ether for 10 min. Atrial cannulation for blood sampling was performed through the jugular vein 5 days before the experiments. In both protocols, blood samples were taken immediately before (0), and 5, 15 and 20 min after the beginning of stress exposure. Ours results showed attenuated endocrine and metabolic responses to ether exposure in the maternal separation (MS) group compared to the control group. The measured metabolic parameters, plasma glucose, prolactin, lactate, and insulin secretion, were 32%, 55%, 41%, 73% lower (P < 0.01), respectively, in MS than in control animals. On the other hand, the endocrine and metabolic stress responses to restraint exposure were not affected by maternal separation. There was no difference between the MS and the control groups in any of the parameters studied. Our data demonstrated that early life experiences affect the hormonal systems beyond the hypothalamic–pituitary–adrenal axis, such as the central neuronal pathways, and their activities related to hormonal and metabolic responses to stress in adulthood. More importantly, these modifications were specific, but dependent on stress situation affecting mainly the circuitry related to the stress response to ether exposure.