Marta Volosin

Chronic stress-induced changes in locus coeruleus neuronal activity

Locus coeruleus (LC) activity was assessed in rats exposed to either acute or chronic stress. After one immobilization session, the number of spontaneously active LC neurons dramatically decreased. On the other hand, repeated restraint sessions enhanced noradrenergic (NA) transmission and the inhibitory effect of clonidine (CLON) was greater on these cells than in those of controls. These results bear on the adaptive changes in the NA system following acute or chronic stress.

Metyrapone pretreatment prevents the behavioral and neurochemical sequelae induced by stress

In the present study, we examined the effect of metyrapone, an inhibitor of corticosterone (CS) synthesis, on the behavioral and neurochemical sequelae induced by a brief restraint session. A 15-min stress period induced an anxiogenic-like behavior on the elevated plus-maze (EPM), which was reversed with metyrapone (75 mg/kg i.p.) injected 3 h prior to the stress event. It was further demonstrated that metyrapone pretreatment normalized the decrease in maximal chloride uptake following GABA stimulation observed in brain cortex tissue obtained from animals exposed to both restraint and the EPM. In addition, plasma CS levels were assessed both after restraint and following EPM exposure. Furthermore, the administration of both CS (2.5 mg/kg s.c. at a dose that mimics CS levels induced by restraint) or dexamethasone (DEXA, 1.25 microg/kg s.c) resulted in an anxiogenic response in the EPM comparable to that induced by restraint. Taken together, all these evidence suggest that CS released in response to stress seems to be associated with functional changes at the GABAergic supramolecular complex which could underlie the enhanced anxiety observed following the exposure to an aversive experience.

Characterization and functional significance of glucocorticoid receptors in patients with major depression: modulation by antidepressant treatment

Hyperactivity of the hypothalamic pituitary adrenal (HPA) axis in patients with major depression is one of the most consistent findings in biological psychiatry. Experimental data support the idea that glucocorticoid-mediated feedback via glucocorticoid receptors (GR) is impaired in major depression. The aim of the present work was to assess the putative changes in GR density of peripheral blood mononuclear cells (PBMCs) in a group of patients with major depression and to determine modulation of these GR sites by antidepressant treatment. In addition, susceptibility of PBMCs to glucocorticoid effects was also studied using a functional end-point analysis in vitro, such as cortisol inhibition of mitogen-induced lymphocyte proliferation. Cortisol levels were also measured before and after dexamethasone suppression test (DST). The results showed a decrease in GR density in depressed patients compared with healthy subjects, mainly in those patients that showed basal cortisol levels in the upper normal range and were refractory to DST. Regarding the functional significance of this variation, two representative groups emerged from our study: a) free-medication patients with GR function comparable to healthy controls, and b) patients showing diminished GR activity. These results suggest a lack of relationship between GR density and cortisol-induced inhibition of lymphocyte proliferation. Patients treated with different antidepressant drugs showed a marked increase in the number of GR sites per cell compared to non-treated. Interestingly, this increase was even higher than in normal subjects. Hence, restoration of GR density after an efficient antidepressant treatment could be an index of an effective modulatory action of drugs on GR expression and highlights the possibility that GR levels might be used as markers of a successful treatment.

Prior exposure to a brief restraint session facilitates the occurrence of fear in response to a conflict situation: behavioral and neurochemical correlates.

The influence of two different stressors on the behavioral and neurochemical responses to a subsequent exposure to the elevated plus maze (EPM) was examined. Rats were submitted to either a 15-min restraint period or to a 15-min forced swimming test (FS) and one day later exposed to the EPM. Animals with early restraint exhibited a significant decrease in the percent time spent and in the number of entries on the open arms. In addition, restraint induced a reduction in the total number of entries. An identical behavior in the EPM was observed between unstressed rats and those exposed to a previous swimming experience. As a humoral index of stress, corticosterone (CS) secretion in response to each stressor was evaluated. A similar increase of CS release was observed following each aversive stimulus. Exposure to both restraint and EPM decreased the cortical chloride uptake following GABA stimulation. Similar values of chloride flux were obtained from animals submitted to either restraint but without subsequent exposure to the EPM, exposed only to the EPM, or without any manipulation (controls). These findings are discussed in terms of a facilitated behavioral and neurochemical response to a fearful situation following an early and brief restraint experience.

Glucocorticoid and Mineralocorticoid Receptors Are Involved in the Facilitation of Anxiety-Like Response Induced by Restraint

In previous studies, we have shown that male Wistar rats exposed to a single inescapable stressor session (15 min restraint) exhibited 24 h later an anxiogenic-like behavior in the elevated plus-maze (EPM), which was reversed by inhibition of corticosterone (CS) synthesis with metyrapone (75 mg/kg i.p.) 3 h before stress. Since CS binds to two central corticosteroid receptors, the mineralocorticoid (MR) and the glucocorticoid (GR) receptors, involvement of MR and GR in the modulation of anxiogenic responses was assessed in the EPM. Administration of the GR agonist dexamethasone (Dex, 1.25 µg/kg s.c.) to metyrapone-pretreated rats 1 h before restraint restored the anxiogenic-like response induced by the stressor. Removal of the adrenals also inhibited the anxiogenic-like effect, which was restored by either Dex (1.25 µg/kg s.c.), the MR agonist deoxycorticosterone (0.8 mg/kg s.c.) or CS, the common endogenous agonist of MR and GR (5 mg/kg s.c.) administered 1 h before stress. Intracerebroventricular infusion to intact animals 15 min before restraint of either a selective GR antagonist (A-GR, RU 38486, 100 ng/2 µl), a selective MR antagonist (A-MR, RU 28318, 100 ng/2 µl) or a combination of A-GR and A-MR (100 ng of each one/2 µl), abolished the stress-induced anxiogenic-like effect. The present findings indicate that both MR and GR are involved in the long-term CS modulation of the anxiety response induced by restraint. Both receptors mediate CS effects in an independent manner.

Corticosterone influences forced swim-induced immobility

The effect of corticosterone (CS) synthesis inhibition with metyrapone-a blocker of the 11 beta-hydroxylase (150 mg/kg IP)-on immobility time during the forced swim test was recorded. Immobility time was measured during a 15-min forced swim (test). Twenty-four hours later rats were subjected to an additional 5 min forced swim (retest). In one experiment, metyrapone or vehicle was administered 3 h before the initial test, while CS (0, 5, 10, or 20 mg/kg SC) was administered 1 h prior to the initial test. Metyrapone significantly reduced immobility time during both test and retest. This effect was reverted in a dose-dependent fashion by CS. In a second experiment, animals exposed to the initial test 24 h before were injected with metyrapone or vehicle 3 h before the retest, while CS (0, 10, or 20 mg/kg SC) was administered 1 h prior the retest. Metyrapone, administered before the retest, reduced immobility time and CS partially reverted metyrapone effect. In another group of animals, serum CS concentrations were evaluated before and after test and retest. In vehicle groups, the high immobility time during test and retest was associated with high CS serum concentrations poststress. In animals receiving metyrapone prior to the initial test, the reduced immobility time was related to low levels of CS after the test and an attenuated secretion following the retest. Moreover, CS (20 mg/kg) and metyrapone+CS groups had high CS levels before the test, which remained high 2 h after the test, although after the retest, both groups showed a pattern of CS secretion similar to that observed in vehicle animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Chronic stress attenuation of α2-adrenoceptor reactivity is reversed by naltrexone

Abstract Low doses of clonidine (50–100 μg/kg IP) evoke a clear dose-dependent hypoactivity response. Seven daily immobilization sessions prevented the motor activity decrease induced by clonidine. On the contrary, a single stress failed to modify clonidine response. Pretreatment with naltrexone (2 mg/kg IP) fully antagonized the attenuating effect induced by chronic stress on clonidine sedative action. These evidences suggest that chronic but not acute stress reduces the reactivity of α 2 -adrenoceptors involved in clonidine-induced sedation. In addition, a regulatory mechanism of endogenous opioids seems to participate on α 2 -adrenoceptors adaptative changes.

Glucocorticoid receptors in lateral septum are involved in the modulation of the emotional sequelae induced by social defeat.

The current research studied the behavior adopted in the elevated plus maze (EPM) of rats previously subjected to a social defeat using the resident-intruder paradigm. One day after defeat, intruder animals exhibited an anxiogenic-like behavior in the EPM. In addition, we also evaluated the role of the corticosteroid receptor system (minerlocorticoid - MR - and glucocorticoid - GR - receptors) from the lateral septum (LS) on the anxiety generated by social defeat. The LS is an area of the aversive circuitry that is preferentially activated in passive defensive postures, and participates - together with other brain areas - in the modulation of aversive states. Intruder animals were infused into the LS with the MR or GR antagonist (ZK 91587 and RU 38486, respectively) and then submitted to social stress. All rats were tested in the EPM 1 day later. Only the administration of the GR antagonist, but not the MR antagonist, into the LS normalized the anxiogenic response induced by defeat. Furthermore, we examined whether a single injection of corticosterone (CS) could induce the same influence on the behavior in the EPM as that observed after social defeat. Moreover, we explored the effect of local infusions of MR or GR antagonists into the LS on the behavior exhibited by CS-treated rats in a subsequent EPM exposure. CS administration also exerted an increased anxiogenic-like behavior, which was normalized only by the local infusion of the GR antagonist. Based on these findings, we suggest that CS secreted by emotionally relevant stimuli acting via GR in LS plays an important role in the modulation of the emotional sequelae induced by social defeat.

Opioid involvement in the adaptive change of 5-HT1 receptors induced by chronic restraint.

Rats immobilized for 2 h daily for 7 days showed an increased behavioral response (forepaw treading and hind-limb abduction) to 5-methoxy-N,N-dimethyltryptamine (5-MeODMT) 24 h after the last stress session. An injection of naloxone before each stress session fully antagonized the increased behavioral reactivity to 5-MeODMT. Treatment with morphine or beta-endorphin associated with each immobilization session for 3 days produced a response to 5-MeODMT higher than that of animals subjected to immobilization only. Chronic immobilization for 7 days did not affect the shaking behavior induced by 5-hydroxytryptophan (5-HTP) 24 h after the last restraint session. These findings suggest that chronic stress may induce a selective adaptive change of the 5-HT1 site and activate an opioid mechanism that is most likely to be involved in the development of this adaptive change.

Corticosterone is involved in foot shock-induced inactivity in rats.

Inescapable shock (IS) exposure induces behavioral inactivity, related to behavioral alterations in subsequent tests (i.e., escape failure, and inactivity during shuttle box task). Metyrapone (150 mg/kg, IP), a corticosterone (CS) synthesis inhibitor, administered 3 h prior to IS reduced inactivity during this aversive experience. Forty-eight hours later, when these rats were submitted to a shuttle box task, a reduction in both escape failure and inactivity was observed. These effects were reversed by CS (20 mg/kg, SC) and dose dependent of the synthetic glucocorticoid dexamethasone, both administered 1 h before IS. When metyrapone was administered 3 h before the shuttle box task to IS-exposed animals, escape failures and inactivity were markedly reduced. This effect was subsequently reversed by CS. The dynamics of changes in serum CS concentrations after both IS and shuttle box task paralleled behavioral changes. Animals injected with metyrapone before IS, which displayed active behavior, showed serum CS levels stable at their basal levels after shock, and their secretion pattern was quite attenuated after the shuttle box task, whereas vehicle-, CS alone-, and metyrapone + CS-injected animals showed higher serum CS concentrations post-IS, which slowly decreased to their corresponding basal levels. CS secretion after the shuttle box task was similar for the three groups: it had the same magnitude as after IS, though the decrease was faster. In all groups, animals displayed passive behavior. These results indicate that glucocorticoids are involved in the onset and expression of passive behaviors induced by uncontrollable stressors. Therefore, it is possible to suggest a functional relationship between CS released by exposure to inescapable stressor and the behavioral strategies adopted by rats under this stressful condition.