Glucocorticoids are involved in the long-term effects of a single immobilization stress on the hypothalamic-pituitary-adrenal axis.

We have previously observed that a single exposure to a severe stressor such as immobilization (IMO) induces long-lasting desensitization of the responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis to the same stressor that enhances rather than dissipates with time (days). As this desensitization of the HPA axis was not observed in response to a novel stressor, we suggested this might be a particular type of learning linked to severe stressful situations. Taking into account the evidence that glucocorticoids are involved in learning and memory, the present study addresses the role of glucocorticoids in the induction of long-term effects of an acute exposure to IMO. Three different experimental approaches were used: (i) blockade of stress-induced corticosterone release by using adrenalectomized rats supplemented with a low dose of corticosterone in the drinking saline (ADX+B); (ii) blockade of corticosterone synthesis during the first exposure to IMO with the 11-beta-hydroxylase inhibitor metyrapone (200 mg/kg); and (iii) administration of the glucocorticoid receptor antagonist RU486 (100 mg/kg). Previous exposure to IMO resulted in an enhanced post-stress recovery of the HPA response to the same stressor 1 week later. These long-term effects of IMO were blocked in ADX+B rats, were partially reduced in metyrapone-treated rats and only modestly affected by RU486 administration. These data suggest that glucocorticoids play a partial role in the induction of long-term effects of IMO on the HPA responsiveness to the same stressor, although the weak effect of RU486 suggests that non-classical corticosteroid receptors may be involved. The role of glucocorticoids in the expression of the phenomenon is suggested by the full blockade of the phenomenon in ADX+B rats, but further studies are needed. As blockade of corticosterone synthesis only partially blunted the long-term effect of IMO, it appears that full induction of the long-term effects of acute exposure to IMO on the HPA axis is only achieved by the concerted action of several endocrine (or neurochemical) factors.

Potentiation of glucocorticoid release does not modify the long-term effects of a single exposure to immobilization stress.

RATIONALE: Previous work has shown that a single exposure of rats to a severe stressor (immobilization, IMO) results, days to weeks later, in a reduced response (desensitization) of the hypothalamic-pituitary-adrenal (HPA) axis to a second exposure to the same stressor. OBJECTIVES: In the present work, we studied the influence of both length of exposure to IMO and circulating levels of corticosterone on the first day on the degree of desensitization of two sets of physiological variables: HPA hormones and food intake. METHODS: Rats were given SC saline or ACTH administration and then exposed to IMO for 0, 1 or 20 min. Seven days later, all rats were exposed to 20 min IMO. HPA response was followed on both experimental days by repeated blood sampling and food intake was measured on a 24-h basis. RESULTS: Both ACTH administration and IMO activates the HPA axis and IMO reduced food intake for several days. A single previous experience with IMO enhanced the post-IMO return of HPA hormones to basal levels on day 8 and reduced the degree of anorexia. The protective effect of previous IMO on food intake was independent of, whereas that on HPA activation was positively related to, the length of exposure on day 1. Concomitant ACTH administration on day 1 did not modify the observed effects. CONCLUSIONS: Long-term protective effects of a single exposure to IMO are observed even with a brief exposure, but they are not potentiated by increasing corticosterone levels during the first exposure.

Behavioral, neuroendocrine and neurochemical effects of the imidazoline I2 receptor selective ligand BU224 in naive rats and rats exposed to the stress of the forced swim test.

RATIONALE: There is evidence for alterations in imidazoline(2) (I(2)) receptor density in depressed patients. Selective I(2) receptor ligands modulate central monoamine levels and activate the hypothalamo-pituitary-adrenal (HPA) axis and may have potential as antidepressants. OBJECTIVES: To study the behavioral effects of the selective I(2) receptor ligand BU224 in the rat forced swim test (FST) and its effects on the HPA axis and central monoaminergic responses. METHODS: Rats received saline or BU224 (10 mg/kg IP) 24, 18 and 1 h prior to 15 min exposure to the FST. Saline- and BU224-treated non-stressed groups were included. Time spent immobile, struggling and swimming calmly was measured. Plasma adrenocorticotrophic hormone (ACTH) and corticosterone levels 90 min post-BU224 were measured in addition to tissue levels of monoamines and metabolites in the frontal cortex, hippocampus and hypothalamus. RESULTS: Administration of BU224 significantly reduced immobility and increased mild swimming without affecting struggling. Exposure to the FST significantly increased plasma ACTH and corticosterone levels. BU224 administration also increased ACTH and potentiated the ACTH response to FST with no effect on corticosterone. BU224 administration significantly increased frontal cortex 5-hydroxytryptamine (5-HT) levels and decreased 5-HT turnover in the frontal cortex and hypothalamus of rats exposed to FST. In non-stressed rats, BU224 decreased 5-HT turnover in the hippocampus and hypothalamus and decreased norepinephrine turnover in the frontal cortex. CONCLUSIONS: The selective I(2) receptor ligand BU224 reduces immobility of rats in the FST, indicative of antidepressant-like activity. This effect is accompanied by alterations in HPA axis and central monoaminergic activity.

Is repeated exposure to immobilization needed to induce adaptation of the hypothalamic-pituitary-adrenal axis? Influence of adrenal factors.

We have previously observed that a single exposure to a severe stressor such as immobilization in wooden boards (IMO) resulted in a faster return of plasma corticosterone (and to a lesser extent of ACTH) to basal activity when the rats were exposed again to the same stressor. In addition, the effect enhanced with time (days) elapsed between the two exposures. These data raised the question of to what extent adaptation of the hypothalamic-pituitary-adrenal (HPA) axis to repeated stress might be, at least partially, explained by the time elapsed between the two exposures rather than by daily repetition of the stressor. To answer this question and the role of glucocorticoids in the process, we studied the effects of single versus repeated exposure to IMO on the HPA response to the same stressor in both sham-operated and adrenalectomized rats maintained with corticosterone in their drinking saline (ADX+B). In sham rats, daily exposure to 20 min IMO for 9 days resulted in a decrease of the ACTH response to the stressor and a faster return of corticosterone to basal levels in the post-stress period. Similar effects were observed with a single session of IMO 8 days before. In ADX+B rats, a reduction of the ACTH response to the stressor was observed in repeated IMO rats but not in single IMO rats. The present results suggest that (i) in sham rats, a single exposure to IMO can induce a degree of adaptation of the HPA response to the same stressor applied days later that is very similar to that caused by repeated exposure to the situation; (ii) stress-induced release of glucocorticoids (or other adrenal factors) is not mandatory for the development of adaptation of the HPA axis to repeated stress, but may be involved in the long-term effects of a single exposure to stress.

Long-term effects of a single exposure to immobilization: a c-fos mRNA study of the response to the homotypic stressor in the rat brain.

A single exposure to a severe emotional stressor such as immobilization in wooden boards (IMO) causes long-term (days to weeks) peripheral and central desensitization of the hypothalamic-pituitary-adrenal (HPA) response to the same (homotypic) stressor. However, the brain areas putatively involved in long-term desensitization are unknown. In the present experiment, adult male rats were subjected to 2 h of IMO and, 1 or 4 weeks later, exposed again to 1 h IMO together with stress-naive rats. C-fos mRNA activation just after IMO and 1 h after the termination of IMO (post-IMO) were evaluated by in situ hybridization. Whereas in most brain areas c-fos mRNA induction caused by the last IMO session was similar in stress-naive (controls) and previously immobilized rats, a few brain areas showed a reduced c-fos mRNA response: ventral lateral septum (LSv), medial amygdala (MeA), parvocellular region of the paraventricular hypothalamic nucleus (pPVN), and locus coeruleus (LC). In contrast, an enhanced expression was observed in the medial division of the bed nucleus stria terminalis (BSTMv). The present work demonstrates that a previous experience with a stressor can induce changes in c-fos mRNA expression in different brain areas in response to the homotypic stressor and suggests that LSv, MeA, and BSTMv may be important for providing signals to lower diencephalic (pPVN) and brainstem (LC) nuclei, which results in a lower physiological response to the homotypic stressor.

Mapping the areas sensitive to long-term endotoxin tolerance in the rat brain: a c-fos mRNA study.

We have recently found that a single endotoxin administration to rats reduced the hypothalamic-pituitary-adrenal response to another endotoxin administration 4 weeks later, which may be an example of the well-known phenomenon of endotoxin tolerance. However, the time elapsed between the two doses of endotoxin was long enough to consider the above results as an example of late tolerance, whose mechanisms are poorly characterized. To know if the brain plays a role in this phenomenon and to characterize the putative areas involved, we compared the c-fos mRNA response after a final dose of endotoxin in animals given vehicle or endotoxin 4 weeks before. Endotoxin caused a widespread induction of c-fos mRNA in the brain, similar to that previously reported by other laboratories. Whereas most of the brain areas were not sensitive to the previous experience with endotoxin, a few showed a reduced response in endotoxin-pretreated rats: the parvocellular and magnocellular regions of the paraventricular hypothalamic nucleus, the central amygdala, the lateral division of the bed nucleus and the locus coeruleus. We hypothesize that late tolerance to endotoxin may involve plastic changes in the brain, likely to be located in the central amygdala. The reduced activation of the central amygdala in rats previously treated with endotoxin may, in turn, reduce the activation of other brain areas, including the hypothalamic paraventicular nucleus.

Stress-induced activation of the immediate early gene Arc (activity-regulated cytoskeleton-associated protein) is restricted to telencephalic areas in the rat brain: relationship to c-fos mRNA.

Arc is an effector immediate early gene whose expression is induced in situations of increased neuronal activity. However, there is no report on the influence of stress on Arc expression. Here, we compared the induction of both c-fos and Arc mRNAs in the brain of rats exposed to one of three different stressful situations: novel environment, forced swimming and immobilization. An absent or weak c-fos mRNA signal was observed in control rats, whereas those exposed to one of three stressors showed enhanced c-fos expression in a wide range of brain areas. Constitutive Arc expression was observed in some areas such as cortex, striatum, hippocampus, reticular thalamic nucleus and cerebellar cortex. In response to stressors, a strong induction of Arc was observed, but the pattern was different from that of c-fos. For instance, activation of Arc but not c-fos was observed in the nucleus accumbens after immobilization and in the hippocampus after novel environment. No Arc induction was observed in diencephalic and brainstem areas. The present data show that Arc has a neuroanatomically restricted pattern of induction in the brain after emotional stress. Telencephalic activation suggests that a more intense induction of synaptic plasticity is occurring in this area after exposure to emotional stressors.

Long-term effects of a single exposure to immobilization stress on the hypothalamic-pituitary-adrenal axis: transcriptional evidence for a progressive desensitization process.

In the present work we have characterized the long-term influence of a single exposure to the stress of immobilization (IMO) on the hypothalamic-pituitary-adrenal (HPA) axis of adult rats. Rats without prior stress (control) and rats exposed to IMO for 2 h on day 1 (IMO+4wk) or on day 21 (IMO+1wk) were killed on day 28, either without stress (basal), immediately after IMO for 1 h (IMO), or 1 h after termination of IMO (post-IMO). IMO caused a strong activation of c-fos mRNA and corticotropin-releasing factor (CRF) and vasopressin (AVP) heteronuclear RNA (hnRNA) in the paraventricular nucleus of the hypothalamus in control rats; this activation was essentially maintained in the post-IMO period. The overall AVP hnRNA response to day 28 stress was not affected by prior stress. Post-IMO c-fos mRNA and CRF hnRNA levels were lower in previously stressed rats, as compared with controls. Whereas the effect of prior IMO on both peripheral HPA hormones and c-fos mRNA was maximal in IMO+1wk rats, the effect of prior stress on CRF hnRNA was only observed in IMO+4wk rats. The present data indicate that prior single IMO triggers a process of desensitization of the HPA responsiveness to IMO over the course of the following weeks. Although the various components of the HPA axis were modified in the same direction, a clear temporal dissociation was found among them, revealing the fine tuning of stress-induced activation of the HPA axis.