Both the magnitude and the duration of the hormonal stress response change dramatically during neonatal development and aging as well as with prior experience with a stressor. However, surprisingly little is known with regard to how pubertal maturation and experience with stress interact to affect hypothalamic-pituitary-adrenal axis responsiveness. Because adolescence is a period of neurodevelopmental vulnerabilities and opportunities that may be especially sensitive to stress, it is imperative to more fully understand these interactions. Thus, we examined hormonal and neural responses in prepubertal (28 d of age) and adult (77 d of age) male rats after exposure to acute (30 min) or more chronic (30 min/d for 7 d) restraint stress. We report here that after acute stress, prepubertal males exhibited a significantly prolonged hormonal stress response (e.g. ACTH and total and free corticosterone) compared with adults. In contrast, after chronic stress, prepubertal males exhibited a higher response immediately after the stressor, but a faster return to baseline, compared with adults. Additionally, we demonstrate that this differential stress reactivity is associated with differential neuronal activation in the paraventricular nucleus of the hypothalamus, as measured by FOS immunohistochemistry. Using triple-label immunofluorescence histochemistry, we found that a larger proportion of CRH, but not arginine vasopressin, cells are activated in the arginine vasopressin in response to both acute and chronic stress in prepubertal animals compared with adults. These data indicate that experience-dependent plasticity of the hypothalamic-pituitary-adrenal neuroendocrine axis is significantly influenced by pubertal maturation. (Endocrinology 147: 1664 -1674, 2006)

The pubertal maturation of the hypothalamic-pituitary-adrenal (HPA) axis has received relatively little experimental attention. The present set of experiments sought to extend our understanding of the pubertal stress response by measuring corticotropin (ACTH), corticosterone, and testosterone levels in prepubertal and adult male rats exposed to a single 30-min session of restraint stress. We show that ACTH and corticosterone levels in prepubertal males take significantly longer to return to baseline after termination of the stressor compared to adults. These data indicate that prepubertal males demonstrate a more prolonged stress response compared to adults after a single acute stressor with both psychogenic and neurogenic components. As testosterone has been shown to reduce the stress response in adulthood, we next investigated whether the relatively low levels of circulating testosterone in prepubertal males mediated their protracted stress response. Data collected from additional experiments revealed that prepubertal males treated with adult-like physiological levels of testosterone still exhibited an extended stress response compared to similarly treated adults. These results indicate that the stress response demonstrated by adult males cannot be mimicked or activated in prepubertal males by mere supplementation of testosterone. Thus, we conclude that the HPA neuroendocrine axis is further shaped during pubertal development to allow for the emergence of a more tightly regulated stress response in adulthood.

Numerous studies have established that adolescence is marked by substantial changes in stress reactivity and hippocampal function. Glucocorticoid receptors (GRs) in the hippocampus are imperative in corticosterone-dependent gene transcription when glucocorticoid levels are relatively high, such as during periods of stress. As reported previously, in reaction to acute stress, prepubertal animals show a significantly more protracted corticosterone response compared to adults. Chronic stress, however, results in a higher peak response, but a faster return to baseline in prepubertal compared to adult animals. Thus, depending on the developmental stage and experience of the animal, the hippocampus is exposed to different concentrations and durations of corticosterone. The present set of experiments assessed the effects of acute or repeated stress on GR mRNA expression in the dorsal and ventral hippocampal formation either before or after pubertal maturation in male rats. We found that acute stress results in a significant decrease in GR mRNA in the CA1 pyramidal cell layer and dentate gyrus in the dorsal and ventral hippocampal formation of both prepubertal and adult males. In response to repeated stress, we found no differences in GR expression in either the dorsal or ventral hippocampus. Thus, despite the dramatic differences in corticosterone concentration following stress at these two developmental stages, the stress-induced changes in GR expression in the hippocampus before and after pubertal maturation were more similar than different. These data point to a dissociation between differential stress-induced corticosterone responses and regulation of hippocampal GR levels in prepubertal and adult animals.