Blunted neurobiological reactivity and attentional bias to threat underlie stress-related disorders in women survivors of intimate partner violence.

BACKGROUND: Intimate partner violence (IPV) alters women's neurobiological stress response systems. We propose that individual differences early in the attentional processing of threats are associated with these neurobiological mechanisms and contribute to mental illness in this population. METHODS: We assessed attentional bias in relation to threat (AB) in women survivors of IPV (n = 69) and controls (n = 36), and examined overall cortisol secretion using hair cortisol (HC), and stress responsiveness measuring salivary cortisol and α-amylase (sAA) before (T0), and after (T1, T2) an acute psychosocial stress task (Trier Social Stress Test). We used repeated-measures ANCOVAs to explore the associations between Group (IPV, control) and AB with acute stress response, and regression models to examine the associations with mental health symptoms. RESULTS: There were no between-group differences in HC levels. An interaction between Group and AB was found regarding cortisol reactivity (p < 0.05). IPV women with threat avoidance AB showed a blunted cortisol response compared to controls and to IPV participants with threat vigilance AB. The association between sAA reactivity and the interaction between Group, AB, and time approached significance (p = 0.07), with a trend to lower sAA levels particularly in IPV women with threat avoidance AB. Group and cortisol reactivity were associated with symptoms of depression, generalized anxiety, and post-traumatic stress disorder (8-20% explained variance). CONCLUSIONS: Threat avoidance AB is associated with blunted acute cortisol response among women exposed to chronic stress (IPV). Experiencing IPV and acute cortisol response appear to be clearly implicated in long-term mental health problems.

Design and optimization of a single-use optical sensor based on a polymer inclusion membrane for zinc determination in drinks, food supplement and foot health care products.

A single-use optical sensor was designed for Zn(II) determination based on the immobilisation of the colorimetric reagent 2-acetylpyridine benzoylhydrazone (2-APBH) in a polymer inclusion membrane (PIM) adhered on the surface of an inert rectangular strip of polyester (Mylar). Different components for the membrane preparation were tested and those resulting in membrane with good appearance, proper physical and optical properties and ease of preparation were selected. Factorial design 23 with three replicates of the central point was applied for the optimisation of the membrane composition. The optimal composition consisted of 2.5 g of poly(vinyl chloride) (PVC), 4 mL of tributyl phosphate (TBP) and 0.04 g of 2-APBH. The optode showed a linear dynamic range from 0.03 (detection limit) to 1 mg L-1 of Zn(II) ions with a response time of 30 min in aqueous solution at pH 6 and a relative standard deviation of 3.90% for 0.4 mg L-1 of Zn(II). The sensor exhibited good selectivity to Zn(II) over other commonly ions. It was successfully applied to the determination of Zn(II) in a water certified reference material, spiked tap water, vitamin-mineral drink, food supplement and foot health care products, as contribution to the concern about this heavy metal due to its significant role in many biological and physiological processes although toxicant at high doses.

An approach to an acute emotional stress reference scale. / Aproximacion a una escala de referencia de estres emocional agudo.

INTRODUCTION: The clinical diagnosis aims to identify the degree of affectation of the psycho-physical state of the patient as a guide to therapeutic intervention. In stress, the lack of a measurement tool based on a reference makes it difficult to quantitatively assess this degree of affectation. AIM: To define and perform a primary assessment of a standard reference in order to measure acute emotional stress from the markers identified as indicators of the degree. SUBJECTS AND METHODS: Psychometric tests and biochemical variables are, in general, the most accepted stress measurements by the scientific community. Each one of them probably responds to different and complementary processes related to the reaction to a stress stimulus. The reference that is proposed is a weighted mean of these indicators by assigning them relative weights in accordance with a principal components analysis. RESULTS: An experimental study was conducted on 40 healthy young people subjected to the psychosocial stress stimulus of the Trier Social Stress Test in order to perform a primary assessment and consistency check of the proposed reference. The proposed scale clearly differentiates between the induced relax and stress states. CONCLUSIONS: Accepting the subjectivity of the definition and the lack of a subsequent validation with new experimental data, the proposed standard differentiates between a relax state and an emotional stress state triggered by a moderate stress stimulus, as it is the Trier Social Stress Test. The scale is robust. Although the variations in the percentage composition slightly affect the score, but they do not affect the valid differentiation between states.

TITLE: Aproximacion a una escala de referencia de estres emocional agudo.Introduccion. El diagnostico clinico persigue identificar el grado de afectacion del estado psicofisico del paciente como orientacion hacia la intervencion terapeutica. En el estres, la falta de un instrumento de medicion por comparacion con una referencia dificulta la valoracion cuantitativa del nivel de afectacion. Objetivo. Definir y hacer una primera validacion de un patron de referencia para la medida del estres emocional agudo a partir de marcadores identificados como indicadores del nivel. Sujetos y metodos. En general, las medidas mas solidas y aceptadas de estres por la comunidad cientifica son los test psicometricos y las variables bioquimicas. Cada uno de ellos responde probablemente a procesos distintos y complementarios de la reaccion frente a un estimulo estresante. La referencia que se propone es una media ponderada de estos indicadores, asignandoles pesos relativos de acuerdo con un analisis de componentes principales. Resultados. Para una primera aproximacion y verificacion de coherencia de la referencia propuesta, se ha utilizado un estudio experimental con una muestra de 40 jovenes sanos sometidos al estimulo estresante psicosocial del Trier Social Stress Test. La escala propuesta diferencia netamente entre los dos estados con distintos niveles de estres inducido. Conclusiones. Aceptando la subjetividad de la definicion, y a falta de una validacion posterior con nuevos datos experimentales, el patron propuesto diferencia entre un estado de relax y uno de estres emocional generados con un estimulo estresante moderado, como es el Trier Social Stress Test. La escala es robusta, ya que variaciones en la composicion porcentual repercuten ligeramente en la puntuacion, pero no en la diferenciacion valida entre estados.

IL-6 and TNF-α in unmedicated adults with ADHD: Relationship to cortisol awakening response.

There is preliminary evidence that the immune system's cytokines may have impact on ADHD in children. Nevertheless, studies exploring the possible role of pro-inflammatory cytokines in adults with ADHD are lacking. This study aimed to assess differences in serum IL-6 and TNF-α between patients and controls and their possible relationship to resting cortisol. 108 adults with ADHD (DSM-IV), 44 inattentive and 64 combined, age ranging between 18 and 55 years, and 27 healthy controls were included. Major psychiatric disorders and organic comorbidities were excluded. Serum samples for IL-6 and TNF-α and salivary samples to assess cortisol awakening response were collected on the same day. Analysis of variance was applied to study differences in IL-6 and TNF-α between groups. Pearson correlations were used to study associations between IL-6, TNF-α, and CAR. There were no significant differences in serum IL-6 or TNF-α levels between patients and controls or between combined and inattentive patients. Negative associations between IL-6 (r=-0.386, p=0.020), TNF-α (r=-0.372, p=0.023) and cortisol awakening response were found in the inattentive subtype, whereas no association was seen in the combined subtype. A negative correlation between IL-6 and cortisol was also present in the control group (r=-0.44, 0.030). The peripheral pro-inflammatory markers, IL-6 and TNF-α, do not appear to be primarily involved in ADHD in adults, although the role of other inflammatory markers cannot be ruled out. The differences regarding the association between IL-6 and TNF-α and morning cortisol response suggest possible underlying neurobiological differences between the inattentive or combined patients that merit further studies.

Maternal deprivation and adolescent cannabinoid exposure impact hippocampal astrocytes, CB1 receptors and brain-derived neurotrophic factor in a sexually dimorphic fashion.

We have recently reported that early maternal deprivation (MD) for 24 h [postnatal day (PND) 9-10] and/or an adolescent chronic treatment with the cannabinoid agonist CP-55,940 (CP) [0.4 mg/kg, PND 28-42] in Wistar rats induced, in adulthood, diverse sex-dependent long-term behavioral and physiological modifications. Here we show the results obtained from investigating the immunohistochemical analysis of CB1 cannabinoid receptors, glial fibrillary acidic protein (GFAP) positive (+) cells and brain-derived neurotrophic factor (BDNF) expression in the hippocampus of the same animals. MD induced, in males, a significant increase in the number of GFAP+ cells in CA1 and CA3 areas and in the polymorphic layer of the dentate gyrus (DG), an effect that was attenuated by CP in the two latter regions. Adolescent cannabinoid exposure induced, in control non-deprived males, a significant increase in the number of GFAP+ cells in the polymorphic layer of the DG. MD induced a decrease in CB1 expression in both sexes, and this effect was reversed in males by the cannabinoid treatment. In turn, the drug "per se" induced, in males, a general decrease in CB1 immunoreactivity, and the opposite effect was observed in females. Cannabinoid exposure tended to reduce BDNF expression in CA1 and CA3 of females, whereas MD counteracted this trend and induced an increase of BDNF in females. As a whole, the present results show sex-dependent long-term effects of both MD and juvenile cannabinoid exposure as well as functional interactions between the two treatments.

Adolescent pre-exposure to ethanol or MDMA prolongs the conditioned rewarding effects of MDMA.

Adolescents often take ethanol (EtOH) in combination with MDMA (3,4-methylenedioxymethylamphetamine). In the present work we studied the effect of repeated intermittent adolescent pre-exposure to both drugs on the behavioral and neurochemical effects of MDMA in mice. Sixteen days after pre-treatment, the rewarding and reinstating effects of MDMA in the conditioned place preference (CPP) paradigm were evaluated, along with the levels of biogenic amines, basal motor activity and corticosterone response to different challenges. Pre-exposure to EtOH, MDMA or EtOH+MDMA did not affect the CPP induced by 10mg/kg of MDMA. However, adolescent exposure to EtOH or MDMA increased the duration of the conditioned rewarding effects of MDMA. Following extinction of the CPP, a priming dose of 5mg/kg of MDMA elicited reinstatement in all the groups, with the duration of this reinstated CPP being longer in mice pre-treated with MDMA. After reinstatement, an increase in monoamine levels was observed in mice pre-exposed to EtOH (DA, DOPAC and 5-HT in the striatum and 5-HIAA in the cortex and hippocampus) or MDMA (5-HT in the hippocampus). Basal motor activity and basal levels of corticosterone were not affected by any of these pre-treatments, but the group pre-exposed to MDMA showed higher levels of corticosterone in response to the administration of 10mg/kg of MDMA. Behavioral and hormonal effects of adolescent exposure to MDMA were reversed by co-administration of EtOH. Our results suggest that exposure to EtOH or MDMA during adolescence prolongs the rewarding properties of MDMA.

Mecanismos de susceptibilidad al estrés / Mechanisms of susceptibility to stress

Está bien establecido que las consecuencias fisiológicas y patológicas de la exposición al estrés dependen de las características de la situación estresante, pero también de las diferencias individuales. Entre las primeras, la intensidad de la situación, su duración, los niveles de imprevisibilidad y el grado de control sobre la situación juegan un papel crítico. Respecto a las diferencias individuales, existen rasgos de personalidad que pueden modular el impacto de las situaciones estresantes o determinar el patrón general de respuesta conductual y fisiológica a las mismas. No obstante, la respuesta puede depender de las características del estímulo estresante y ni siquiera los sistemas fisiológicos más característicos de la respuesta al estrés como la liberación de catecolaminas y de glucocorticoides responden de la misma forma. Todo ello, combinado con la existencia de diferencias individuales en la susceptibilidad de determinados sistemas fisiológicos (incluyendo el SNC) a los cambios causados por el estrés, hace improbable la existencia de fenotipos con una generalizada baja o alta vulnerabilidad al estrés. Aunque el origen de esta susceptibilidad diferencial no se conoce en muchos casos, se ha demostrado no solo la importancia de factores genéticos sino también de modificaciones epigenéticas (que pueden ser heredables). Los factores genéticos y ambientales pueden no tener por sí mismos efectos importantes, pero sí la combinación de ambos. Este hecho, junto a las modificaciones epigenéticas, requiere un cambio de paradigma en muchas de las investigaciones genéticas sobre el origen de la susceptibilidad al estrés y a otros procesos patológicos (AU)

It has been well established that the physiological and pathological consequences of exposure to stress not only depend on the characteristics of the stressful situation, but also on the individual differences. Among the former, situation intensity, its duration, level of unpredictability and grade of control on the situation play a critical role. Regarding the individual differences, there are personality traits that may alter the impact of the stressful situations or determine the general pattern of behavior and physiological response to them. However, the response may depend of the characteristics of the stressful stimulus and not even the most characteristic physiological systems of response to stress such as the release of catecholamines and glucocorticoid respond in the same way. All of the above, in combination with the existence of individual differences in susceptibility of certain physiological systems (including the CNS) to the stress-induced changes, makes the existence of phenotypes with a generalized low or high vulnerability to stress unlikely. Although the origin of this differential susceptibility is not known in many cases, not only the importance of genetic factors but also the epigenetic modifications (that may be inheritable) have been demonstrated. The genetic and environment factors may not have important effects by themselves, however their combination does. This fact, together with the epigenetic modifications, require a paradigm change in many of the genetic investigations on the origin of susceptibility to stress and to other pathological conditions (AU)

The brain pattern of c-fos induction by two doses of amphetamine suggests different brain processing pathways and minor contribution of behavioural traits.

Previous studies have shown that amphetamine (AMPH) markedly activates dopaminergic projection areas, together with some important limbic nuclei. However, a global picture of the brain areas activated is lacking and the contribution of the dose of the drug and individual differences to this global brain activation is not known. In the present experiment, we studied in adult male rats the c-fos expression induced by two doses of AMPH (1.5 and 5 mg/kg sc) in a wide range of brain areas, and investigated the possible contribution of novelty-induced activity and anxiety traits. AMPH administration increased Fos+ neurons in an important number of telencephalic, diencephalic and brainstem areas. Interestingly, the ventral tegmental area (VTA) and the dorsal raphe nucleus were activated by the drug, but c-fos expression was restricted to non-dopaminergic and non-serotoninergic neurons, those activated in the VTA being predominantly GABAergic. The use of the factorial analysis, which grouped the areas in function of the correlation between the number of Fos+ neurons observed in each area, revealed three main factors, probably reflecting activation of various relatively independent brain circuits: the first included medial prefrontal cortex regions, most dorsal and ventral striatal subregions and VTA; the second, raphe nuclei; and the third, the different subdivisions of the paraventricular nucleus of the hypothalamus. Other areas such as the central amygdala did not group around any factor. The finding that an important number of activated areas grouped around specific factors is suggestive of activation of partially independent brain circuits. Surprisingly, a minor contribution of novelty-induced activity and anxiety traits on brain activation induced by AMPH was found. It is possible that normal variability in these traits is poorly related to the effects of AMPH or that c-fos expression is not a good tool to reveal such differences.

Marked dissociation between hypothalamic-pituitary-adrenal activation and long-term behavioral effects in rats exposed to immobilization or cat odor.

Exposure of rodents to cats or certain cat odors results in long-term behavioral effects reminiscent of enhanced anxiety that have been considered to model post-traumatic stress disorder. However, other severe stressors such as tail-shock or immobilization in wooden boards (IMO) appear to induce shorter lasting changes in anxiety. In addition, there are controversial results regarding the effects of urine/feces odors. In the present work, we studied in two experiments the relationship between the degree of stress experienced by the animals during exposure to IMO, urine odors or fur odors (as assessed by hypothalamic-pituitary-adrenal activation and plasma glucose) and the short- and long-term behavioral consequences. In the first experiment, rats were individually exposed for 15 min to a novel environment (white large cages) containing either clean cat litter (controls) or litter soiled by cats (urine odors). Half of the rats in each condition were left to freely explore the environment whereas the others were subjected to immobilization (IMO) within the cages. Although ACTH, corticosterone and glucose responses to IMO were much stronger than those to the white cages with clean litter or urine odors (which did not differ from each other), no effect of treatments on anxiety-like behavior in the elevated plus-maze (EPM) were found one week later. However, previous IMO exposure did cause sensitization of the ACTH response to the EPM. In the second experiment, the response to white large cages containing either no odor (controls), litter soiled by cats (urine odor) or a cloth impregnated with cat odor (fur odor) was compared. Urine and fur odors elicited similar ACTH and corticosterone responses that were higher than those of controls, but plasma glucose levels were slightly higher in rats exposed to fur odor. When compared to controls, activity was only diminished in the novel cages containing fur odor. Similarly, fur odor-exposed rats, but not those exposed to urine odor, showed signs of enhanced anxiety in the EPM seven days later, although the ACTH response to the EPM was similar in the three groups. The present data demonstrate: (a) a marked dissociation between the degree of ACTH, corticosterone and glucose responses to stressors and their long-term anxiety-like effects; (b) that the type of cat odor is critical in determining the short-term and long-term physiological and behavioral consequences of exposure; and (c) that plasma ACTH released during brief exposure to the EPM does not appear to reflect anxiety-like behavior.

Previous exposure to immobilisation and repeated exposure to a novel environment demonstrate a marked dissociation between behavioral and pituitary-adrenal responses.

Activation of the hypothalamus-pituitary-adrenal (HPA) axis is presumably related to the degree of novelty and considered to reflect emotional reactivity. Exposure to novel environments can allow us to simultaneously evaluate both behavior and HPA activation and therefore it is an appropriate design to directly study the relationship between both responses. In the present experiment, we studied how previous exposure to a severe stressor (2 h of immobilisation, IMO, 5 days before testing) and repeated exposure to the same novel environment (a holeboard, HB) altered behavioral and HPA response to the HB. Previous exposure to IMO did not alter any behavior during the first exposure to the HB (5 min), but elicited a greater ACTH response as compared to stress-naive rats. However, corticosterone response did not differ between groups, probably because maximum corticosterone levels are never reached before 15-20 min. Repeated exposure of IMO and stress-naive rats to the HB every other day resulted in progressively lower levels of activity/exploration in both groups, whereas the ACTH and corticosterone responses were basically maintained intact over the days. The present results demonstrate a double dissociation between behavior and HPA activation in the HB. First, a single exposure to IMO elicited a long-lasting sensitisation of the HPA axis that apparently was not a direct consequence of fear/anxiety elicited by the novel environment. Second, progressive familiarisation of the animals with a novel environment resulting in apparently lower levels of motivation to explore did not appear to reduce the stressful properties of the situation as evaluated by ACTH release.

Litter size affects emotionality in adult male rats.

The role of natural variations in pre-weaning litter size in rodent adult emotionality and the importance of maternal care as a possible mediating factor have been frequently neglected. To address these issues, maternal behaviour of Sprague-Dawley dams differing in natural number of pups was studied for the first seven postnatal days. Later, adult behaviour of representative male offspring was studied in the elevated plus-maze, the circular corridor, the dark-light box and the forced swimming test. Three groups of offspring were selected in function of the number of littermates: L<10 group (less than 10 pups per dam), L10-15 (between 10 and 15 pups per dam) and L>15 group (more than 15 pups per dam). L<10 litters showed a reduced habituation of activity across time in a circular corridor and as compared to L>15 litters, L<10 litters showed a lower activity during the first 5 min of exposure to the circular corridor. L<10 litters had also higher signs of anxiety in the elevated plus-maze, in comparison to the other two groups. In addition, L<10 litters showed in the forced swimming test reduced struggling and more mild swimming behavior than the other two groups. These abnormalities in L<10 litters are not explained by maternal behavior since they received individually more maternal care than L>15, as assessed by total licking-grooming observed during the whole observation period divided by number of pups. Although previous data from several laboratories have demonstrated that low maternal care is associated with heightened emotionality at adulthood, the present results suggest an important contribution of spontaneous litter size to adult emotional behavior that cannot be explained by concomitant changes in maternal care.

Social stress is as effective as physical stress in reinstating morphine-induced place preference in mice.

RATIONALE: Relapse to drug-seeking in abstinent heroin addicts and reinstatement in experimental animals are observed when exposed to drug-associated stimuli or cues, the drug itself, and stressful events. It has been shown that footshock-induced stress increases the rewarding effects of opiates, delays extinction, and induces the reinstatement of drug-seeking. However, the effects of social stress on the reinstatement of opiate-seeking after extinction has not been studied. OBJECTIVES: The role of physical (restraint and tail pinch) and social (social defeat) stressors on the reinstatement of morphine-induced conditioned place preference (CPP) was evaluated. METHODS: Adult male OF1 mice were conditioned with 10, 20, or 40 mg/kg of morphine or saline. Only morphine-conditioned animals acquired CPP. All mice underwent extinction sessions until the CPP was extinguished. Then, the effects of physical or social stress on the reinstatement of CPP were evaluated. Morphine- and saline-conditioned animals were exposed to the respective stressor or control stress condition immediately or 15 min before reinstatement tests. In experiment 1, animals underwent restraint for 15 min. In experiment 2, animals were exposed to tail pinch or placed in a cage without any manipulation for 15 min. In experiment 3, animals performed an agonistic encounter with an isolated or anosmic mouse or were placed in a cage without any social contact or manipulation. RESULTS: Restraint, tail pinch, and social defeat in an agonistic encounter with an isolated mouse produce the reinstatement of CPP in morphine-conditioned animals. CONCLUSIONS: These data demonstrate that social stress is as effective as physical stress in reinstating morphine-seeking.

Influence of reactivity to novelty and anxiety on hypothalamic-pituitary-adrenal and prolactin responses to two different novel environments in adult male rats.

Since stressor-induced activation of the hypothalamic-pituitary-adrenal (HPA) axis is involved in some stress-related pathologies, much attention has been paid in laboratory animals to the study of the relationship between endocrine, particularly HPA, responsiveness to stressors and other individual characteristics, such as reactivity to novelty and fear/anxiety. In the present study, adult male rats were classified as high or low reactive to novelty (HR versus LR), as a function of the horizontal activity displayed during 30 min in a circular corridor, and as high or low anxiety (HA versus LA) as a function of the time spent in the open arms of the elevated plus-maze. Then, the behavioural and hormonal response to two distinct novel environments (the hole-board and the light-dark) was assessed in the same subjects, using a counterbalanced design. Plasma prolactin, ACTH and corticosterone responses to the hole-board were higher than to the light-dark, a good correlation between the two tests being found for each hormone. Whereas the hormonal response to the novel environments was not affected by anxiety, HR rats showed a consistently higher HPA response than LR rats when the criteria to classify the animals were the activity during the first 15 min in the circular corridor, but not when the activity during the second 15 min was considered. Neither trait affected prolactin response. The present results demonstrate a good within-individual consistency of the endocrine response to novel environments and support the hypothesis of a higher HPA response to stressors for HR versus LR rats. In contrast, no contribution of fear/anxiety to endocrine responsiveness was observed.

Perseverance of exploration in novel environments predicts morphine place conditioning in rats.

Reactivity to novelty has been related to operant drug self-administration but does not seem involved in the conditioned place preference (CPP). To further assess this issue our aims were to investigate: (1) the importance of the initial versus delayed activity in the novel environment to predict the CPP induced by morphine; (2) the separate contribution of trait anxiety in morphine CPP. Male Sprague-Dawley rats were exposed to a circular corridor for 30 min to assess reactivity to novelty and to the elevated plus-maze and the light-dark tests as measures of anxiety and morphine CPP was then studied (three pairings with 5 mg/kg s.c. morphine and three with saline). Delayed activity in the corridor (16-30 min) correlated positively with CPP score, whereas the initial activity (0-15 min) did not. High-responders (HR), those more active during the second half of the corridor, developed morphine CPP in contrast to low-responders (LR). Also, HR and LR did not differ in anxiety nor any plus-maze or light-dark test measure correlated with CPP behaviour. Enhanced vulnerability to develop morphine CPP is predicted by a higher delayed activity in a novel environment, regardless of anxiety.

A single dose of metyrapone caused long-term dysregulation of the hypothalamic-pituitary-adrenal axis in the rat.

There is evidence that metyrapone (MET), apart from its inhibition of 11-beta steroid hydroxylation, may exert some stress-like effects in the brain, including the activation of the hypothalamic-pituitary-adrenal (HPA) axis and the induction of c-fos. Since a single exposure to some stressors has been found to exert long-term effects on the HPA axis, we hypothesized that a single dose of MET (200 mg/kg, s.c.) could exert even stronger effects, due to the combination of its stressful properties with the lack of constrain of the HPA axis by glucocorticoids. Whereas the inhibitory effect of the drug on corticosterone secretion lasted less than 24 h, its stimulatory effect on the HPA axis could be seen for at least 2 days after the injection. Surprisingly, on day 8, an exacerbated HPA response to immobilization stress was observed in MET rats, despite complete normalization of resting levels of HPA hormones. At this time it was also observed, under basal conditions, increased levels of mRNA for CRH and arginin-vasopressin in the parvocellular region of the paraventricular nucleus of the hypothalamus (pPVN), along with reduced mRNA for glucocorticoid receptors in dentate gyrus and hippocampus CA1, but not in pPVN or medial prefrontal cortex. These data suggest that a single MET administration can exert a marked and long-lasting dysregulation of both resting and stress-induced activity of the HPA axis. Thus, attention should be paid to these properties when using the drug to study the functional role of glucocorticoids.

Long-term effects of a single exposure to immobilization on the hypothalamic-pituitary-adrenal axis: neurobiologic mechanisms.

In apparent contrast to previous results from other labs, we have found that a single exposure to a severe stressor such as immobilization (IMO) caused a long-term desensitization of the hypothalamic-pituitary-adrenal (HPA) response to the homotypic stressor. Because such HPA desensitization was not found in response to heterotypic stressors, it seemed at first that we were describing a habituation process already observed after a single experience with the stressor. However, a more detailed analysis revealed two main properties incompatible with the interpretation of the results in terms of habituation: (1) The intensity of desensitization increases over the course of days to weeks with no additional exposures to the stressor, and (2) the degree of desensitization was greater with more severe stressors. The long-term effects were also observed after a single exposure to a high dose of a systemic stressor such as endotoxin but not after insulin-induced hypoglycemia, suggesting that not all severe systemic stressors can induce such long-term desensitization. Because systemic stressors are known to be processed in specific brain areas and because we have found changes in c-fos mRNA response to the homotypic stressor in some brain areas as a consequence of previous experience with IMO, we hypothesize that some severe stressors do not induce long-term desensitization because they are not processed in brain areas sensitive to previous experience with the stressor. The neurochemical mechanisms involved in the induction of long-term effects on the HPA axis are in process, but our results suggest only a partial role of glucocorticoids and NMDA receptors.

The hypothalamic-pituitary-adrenal and glucose responses to daily repeated immobilisation stress in rats: individual differences.

It is accepted that there are important individual differences in the vulnerability to stress-induced pathologies, most of them associated to the hypothalamic-pituitary and sympatho-medullo-adrenal axes, the two prototypical stress-responsive systems. However, there are few studies specifically aimed at characterising individual differences in the physiological response to daily repeated stress in rats. In the present work, male rats were submitted to repeated immobilisation (IMO) stress (1 h daily for 13 days) and several samples were taken at specific days and time points. Animals only subjected to blood sampling procedure served as controls. Daily adrenocorticotropic-hormone (ACTH), corticosterone and glucose responses to immobilisation (that included the post-immobilisation period) progressively declined over the days. In addition, repeated immobilisation resulted in decreased relative thymus weight, increased relative adrenal weight, elevated corticotropin-releasing factor (CRF) mRNA levels in the hypothalamic paraventricular nucleus (PVN), and down-regulation of glucocorticoid receptor gene transcription in hippocampus CA1. However, only CRF mRNA levels in the paraventricular nucleus correlated with the ACTH (on day 1) and corticosterone responses (from day 4-13) to immobilisation. When the animals were classified in three groups on the basis of their plasma ACTH levels immediately after the first immobilisation, individual differences in the ACTH response progressively disappeared on successive exposures to the stressor, whereas those in corticosterone and glucose were more sustained. The present results suggest that there are individual differences in the physiological response to stress that tend to be reduced rather than accentuated by repeated exposure to the stressor. Nevertheless, this buffering effect of repeated stress was dependent on the particular variable studied.

Renal mechanisms involved in stress-induced antinatriuresis and antidiuresis in rats.

The present study was conducted to investigate if changes in sodium and water excretion in stressed animals were due to modifications in the glomerular filtration rate (GFR) and to determine the participation of angiotensin II (Ang II) and alpha and beta-adrenoceptors on sodium and water renal excretion in rats subjected to immobilization stress (IMO). Male Wistar rats (250-300 g) were randomly separated into five different groups and vehicle (0.9% NaCl) via intraperitoneal (i.p.) or propanolol (3 mg/kg i.p.) or captopril (6 mg/kg i.p.) or yohimbine (3 mg/kg i.p.) or prazosin (1 mg/kg i.p.) were injected respectively. During experimental measurements, the animals were kept in metabolic cages for 6 h and sodium, potassium and water renal excretion and saline (1.5% NaCl) and water intake were determined at day 1 (drug effect) and day 7 (drug + IMO effects). GFR was measured by creatinine clearance in control and IMO rats. A stress-induced antinatriuresis and antidiuresis was reversed by alpha 1 and alpha 2-adrenoceptor antagonists, while captopril inhibited only the antidiuresis and propranolol had no effect on either parameter. No differences were observed in creatinine clearance in the studied groups. Since yohimbine blocks alpha 2-adrenoceptors and prazosin blocks alpha 1-adrenoceptors and alpha 2B-adrenoceptors, the stress-induced renal sodium reabsorption mainly could be attributed to alpha 2B-adrenoceptors. The present results indicate that beta-adrenoceptors do not participate in this response and, Ang II only reverses the antidiuresis and shows a slight participation in antinatriuresis. The increment in sodium and water reabsorption caused by IMO occurred without changes in the glomerular filtration rate.

Positive relationship between activity in a novel environment and operant ethanol self-administration in rats.

RATIONALE: The study of individual differences in drug addiction may have important implications both for understanding the etiology of addiction and for strategies for treatment. Activity of rodents in novel environments, presumably related to the novelty-seeking trait in humans, is the primary behavioral feature that is hypothesized to predict a predisposition for drug self-administration by rodents. OBJECTIVES: The aim of this study was to characterize the relationship between motor activity in a novel environment and operant ethanol self-administration using the sucrose-substitution procedure. METHODS: Male Long-Evans rats were exposed to a novel environment for 2 h, and the distance traversed, rearing, and defecation was recorded. After 3 days of forced exposure to ethanol the sucrose-substitution procedure began and lasted for 23 days. Following sucrose substitution the animals were maintained on a schedule of ethanol (10% v/v) self-administration with a fixed ratio 3 (FR3) for 15 days. RESULTS: The activity (distance traversed) in the novel environment was positively correlated with initial ethanol self-administration under the FR3 schedule ( r=0.87) but not with the number of inactive lever presses or active lever presses for ethanol or for sweetened ethanol solutions with lower ratios of response. In contrast, rearing was correlated positively only with the number of inactive lever presses for sucrose. CONCLUSIONS: Motor activity in a novel environment may be related to the acquisition of operant ethanol self-administration only when a given ratio of response is required.

A single lipopolysaccharide administration is sufficient to induce a long-term desensitization of the hypothalamic-pituitary-adrenal axis.

We have previously shown that a single exposure of adult rats to a severe emotional stressor such as immobilization is able to exert a long-term desensitization of the response of the hypothalamic-pituitary-adrenal (HPA) axis to the same stimulus when applied days to weeks later. Surprisingly, the intensity of the effect increased with time elapsed between the two exposures, suggesting that we are dealing with a new type of stress-associated phenomenon. Taking into account the clinical importance of tolerance to endotoxin, in the present study we assessed whether a single exposure to an immunological stressor such as lipopolysaccharide can induce effects similar to those of immobilization. Rats injected with lipopolysaccharide (1 mg/kg) showed a reduction of the response of the corticotropin-releasing factor mRNA in the paraventricular nucleus of the hypothalamus after a new lipopolysaccharide injection 4, but not 2 weeks later. In an additional experiment using a different blood sampling procedure, adrenocorticotropin hormone, corticosterone and tumor necrosis factor-alpha responses were reduced approximately to the same extent by previous experience with lipopolysaccharide either 1 or 4 weeks before. Our data suggest that a previous single exposure to lipopolysaccharide induces a long-lasting tolerance of the HPA axis that likely involves some kind of learning-like brain plasticity.