Endocrine and lymphoproliferative response changes produced by social stress in mice

Daily dyadic resident-intruder encounters and uninterrupted cohabitation in pairs were used to assess the impact of different durations (5 and 15 days) of dominance and subordination experiences on splenic lymphoproliferative responses in male OF1 strain mice. HPA axis activity was assessed by measuring serum corticosterone levels, whereas splenic norepinephrine (NE) content provided a sympathetic activity index. Corticosterone levels in subordinate subjects were generally higher than in their control or dominant counterparts in both treatment paradigms. Corticosterone levels in dominant subjects were lower than in their control counterparts in both. Increasing the duration of treatments generally decreased such titers, especially so in subordinate subjects. No differences were detected in splenic NE content. Animals subjected to social interaction generally showed greater proliferation than their control counterparts. This effect was more pronounced in subordinates than dominants and after longer- rather than short-duration treatments. There was no inverse relation between proliferative responses and the subject's corticosterone levels. While corticosterone may have a general immunomodulating effect, other mediators apparently account for the effects produced by these social stress paradigms on splenic proliferative response.     

The pituitary-adrenal response to novel stimulation and ether stress in young adult and aged rats

Pituitary-adrenal responsiveness to stress induced by either psychological (novelty) or physiological (ether) stimuli was examined in young (3 to 6 months) and old (24 to 27 months) male and female rats. In Experiment 1, subjects were placed in a novel environment for 3 min. Blood samples were collected at 0, 15, 30, 45, and 60 min. In Experiment 2, a blood sample was collected from experimental subjects immediately following 15 min in a novel environment; a basal sample was collected from control subjects. In Experiment 3, basal and stress blood samples were collected from subjects exposed to ether vapors. Blood samples were assayed for plasma corticosterone content. The major findings were: (1) no change with age in either sex in basal corticosterone levels or in the time course of the adrenocortical response; (2) no change with age in males in stress-induced increments in corticoids; (3) no change with age in females in corticoid elevations induced by a mild stressor (Experiment 1), but a decrement in elevations induced by more potent stressors (Experiments 2 and 3).     

Adrenal hormones and brain catecholamines responses to morning and afternoon immobilization stress in rats

The response of adrenal hormones and brain catecholamines to immobilization stress, applied in the morning or in the afternoon, was studied in rats. Immobilization induced greater increments in plasma corticosterone in the morning than in the afternoon; i.e. when the basal levels of this hormone were low rather than high. There was no significant difference between the plasma and adrenal corticosterone levels of the morning and those of the afternoon-stressed rats. Decreases in brain norepinephrine were observed in both groups (a.m. and p.m.) of immobilized rats, but no variations in dopamine concentration were detected. Immobilization led to a decline in the adrenal epinephrine level in the morning but no significant variation was observed in the afternoon-stressed rats.     

Effects of ether stress on prolactin and corticosterone levels in prenatally-stressed male rats as adults

The effect of a potent prenatal stressor on adult levels of prolactin and corticosterone was investigated in male rats. It was found that prenatal stress had no effect on initial levels of either of these two hormones. Ether stress levels of prolactin were significantly lower in prenatally-stressed animals compared to prenatally-nonstressed subjects, whereas corticosterone levels in response to ether stress were only marginally affected in prenatally-stressed animals reared by prepartally-stressed mothers. It is suggested that alterations in endocrine function in the adult animal may result from prenatal stress effects on the development of neural mechanisms that regulate the hormonal response to ether stress.     

Effects of comfort food on food intake, anxiety-like behavior and the stress response in rats

It has been suggested that access to high caloric food attenuates stress response. The present paper investigates whether access to commercial chow enriched with glucose and fat, here referred to as comfort food alters behavioral, metabolic, and hormonal parameters of rats submitted to three daily sessions of foot-shock stress. Food intake, anxiety-like behaviors, and serum levels of insulin, leptin, corticosterone, glucose and triglycerides were determined. The rats submitted to stress decreased the intake of commercial chow, but kept unaltered the intake of comfort food. During the elevated plus maze (EPM) test, stressed rats increased the number of head dipping, entries into the open arms, as well as the time spent there, and decreased the number of stretched-attend posture and risk assessment. These effects of stress were independent of the type of food consumed. Non-stressed rats ingesting comfort food decreased risk assessment as well. Stress and comfort food increased time spent in the center of the open field and delayed the first crossing to a new quadrant. Stress increased the plasma level of glucose and insulin, and reduced triglycerides, although consumption of comfort food increases glucose, triglyceride and leptin levels; no effect on leptin level was associated to stress. The stress induced increase in serum corticosterone was attenuated when rats had access to comfort food. It was concluded that foot-shock stress has an anorexigenic effect that is independent of leptin and prevented upon access to comfort food. Foot-shock stress also has an anxiolytic effect that is potentiated by the ingestion of comfort food and that is evidenced by both EPM and open field tests.     

Cage mate separation in pair-housed male rats evokes an acute stress corticosterone response

Corticosterone (CORT) release from the adrenal glands in response to acutely stressful stimuli is well-characterized, however several non-experimental, environmental stressors can also engender a CORT response. The aim of this study was to investigate an acute activation of the HPA axis in pair-housed animals in response to separation. We observed a rapid significant increase in CORT in the animal remaining in the home cage following cage mate removal, that was not caused by cage opening and transient removal of cage mate. In addition, we examined this response in both control, non-stressed animals and in animals subjected to chronic variable stress (CVS) and found that although basal levels of CORT differed between control and CVS animals, there was no significant difference in the acute CORT levels between the control and CVS animals after separation, indicating that this environmental event is perceived as acutely stressful in both conditions. Furthermore, we examined the time course of CORT activation and found that CORT levels rapidly rise within minutes of separation peaking at 15 min and returning to baseline by 90 min. The results of this study demonstrate that separation can induce an acute stress response in the remaining cage mate measured by increased CORT and should be considered in molecular, behavioral, and electrophysiological studies.     

The nociceptin/orphanin FQ antagonist UFP-101 differentially modulates the glucocorticoid response to restraint stress in rats during the peak and nadir phases of the hypothalamo-pituitary-adrenal axis circadian rhythm

The involvement of nociceptin (N/OFQ) and the nociceptin/orphanin FQ peptide (NOP) receptor in behavior associated with stress and anxiety has been established but their role in the regulation of the hypothalamo-pituitary-adrenal (HPA) axis under conditions of stress has not been fully investigated. We used the selective NOP receptor antagonist UFP-101 to examine the contribution of endogenous N/OFQ to HPA axis control under conditions of restraint stress in the morning and the evening. We found that in the morning during the HPA axis circadian nadir rats exposed to restraint stress in both the presence and absence of UFP-101 exhibited significantly elevated plasma corticosterone at 30 min post-i.c.v. injection compared to the home cage control group. Additionally, rats treated with UFP-101 and exposed to restraint had significantly elevated corticosterone levels at 60 min post-i.c.v. injection compared to all other treatment groups. Interestingly, while there was a significant increase in the expression of CRF mRNA in the paraventricular nucleus (PVN) of rats exposed to restraint stress only, there was no comparable increase in those co-treated with UFP-101. There was no change in the expression of AVP or POMC mRNA in any of the treatment groups. In contrast, when carried out in the evening we observed significantly elevated plasma corticosterone in the vehicle-treated restraint group only at 30 min post-i.c.v. injection. There was no significant difference between the UFP-101-treated restraint group and either of the home cage control groups or the vehicle-treated restraint group. Additionally, in contrast to the morning study, UFP-101 did not prolong glucocorticoid release at the 60 min time-point. These results demonstrate for the first time a differential effect of UFP-101 on restraint stress-induced HPA axis activity characterized by significant prolongation of stress-induced activity in the morning but no significant effect on the response to restraint in the evening.     

Repeated social defeat stress induces chronic hyperthermia in rats

Psychological stressors are known to increase core body temperature (T_c) in laboratory animals. Such single stress-induced hyperthermic responses are typically monophasic, as T_c returns to baseline within several hours. However, studies on the effects of repeated psychological stress on T_c are limited. Therefore, we measured T_c changes in male Wistar rats after they were subjected to 4 social defeat periods (each period consisting of 7 daily 1 h stress exposures during the light cycle followed by a stress-free day). We also assessed affective-like behavioral changes by elevated plus maze and forced swim tests.In the stressed rats, the first social defeat experience induced a robust increase in T_c (+ 1.3 °C). However, the T_c of these rats was not different from control animals during the subsequent dark period. In comparison, after 4 periods of social defeat, stressed rats showed a small but significantly higher (+ 0.2-0.3 °C) T_c versus control rats during both light and dark periods. Stressed rats did not show increased anxiety-like behavior versus control rats as assessed by the elevated plus maze test. However, in the forced swim test, the immobility time of stressed rats was significantly longer versus control rats, suggesting an increase in depression-like behavior. Furthermore, hyperthermia and depression-like behavior were still observed 8 days after cessation of the final social defeat session. These results suggest that repeated social defeat stress induces a chronic hyperthermia in rats that is associated with behavior resembling depression but not anxiety.     

Corticosterone response in group-housed rats exposed to psychogenic stresses in different social conditions

The corticoadrenal stress response of group-housed male Wistar rats was the same in rats stressed alone or with other rats with which they were housed. Thus, individually and grouped, stressed rats showed a similar corticosterone response to both kinds of stress used in the experiment. The effects of the presence of conspecifics on corticoadrenal response to stress could depend on the housing conditions (grouped or isolated) during the days before the experiment.     

Testosterone decrease does not play a major role in the suppression of hippocampal cell proliferation following social defeat stress in rats

Stress of social defeat in rodents is known to have a strong and long-lasting effect on brain, physiology and behavior, which bears similarities with certain human stress related psychopathologies. Previous experiments in this lab showed that social defeat stress suppresses testosterone secretion and causes a lasting desensitization of the serotonergic 5-HT_1A receptors. Testosterone supplementation in socially stressed tree shrews prevented a decrease in hippocampal 5-HT_1A receptor binding. These receptors are hypothesized to play an important role in neurogenesis in this brain structure. We designed the present experiment to test if social defeat reduces hippocampal cell proliferation and neurogenesis in rats and if testosterone supplementation can prevent this reduction. The results indicate that repeated social defeat stress on 5 successive days induces a significant drop in plasma testosterone levels in male rats and suppresses hippocampal cell proliferation 24 h and 3 weeks after the end of the stress period. Testosterone supplementation prevented the social stress induced drop in plasma testosterone levels. The hormone supplementation also reduced the negative effect of stress on hippocampal BrdU labeling at 3 weeks post-defeat. This effect was, however, rather weak and was caused by the tendency of the hormone in itself to suppress proliferation and the failure to fully recover the proliferation rate. Survival of dentate gyrus cells that either proliferated prior to the stress period or 24 h after the last defeat was not affected by the social defeats. Thus the stress-induced lowering of hippocampal cell proliferation is not likely to be caused by transient inhibition of testosterone secretion during social stress.     

Mild prenatal stress in rats is associated with enhanced conditioned fear

We tested the hypothesis that prenatal stress would enhance conditioned fear in adult rats. Pregnant Sprague-Dawley rats were stressed by exposure to a novel environment and subcutaneous injection of saline (0.1 ml 0.9% NaCl) at random times daily from Days 14 to 21 of pregnancy. When compared to adult control (CON) male rats from unmanipulated pregnancies, adult prenatally stressed (PS) male rats showed increased freezing behavior in response to acute footshock as well as increased freezing behavior the next day in the same context, without shock delivery. In another experiment, the gestational stressor was examined for elevations in corticosterone and ACTH. At gestational days (G)15, G17, G19 and G21, maternal and fetal plasma was collected. Analysis showed elevations in corticosterone and ACTH in the PS dams when compared to the CON dams. Additionally, increased corticosterone was found in the PS fetuses when compared to the CON fetuses. Finally, some CON and PS litters were examined for alterations in length of gestation, number of pups born, bodyweight on postnatal day (P)1 and anogenital distance on P1 and differences were not found. In conclusion, our data demonstrate that a mild stressor during gestation, sufficient to raise plasma corticosterone and ACTH, is associated with enhanced conditioned fear during adulthood.     

Predator and restraint stress during gestation facilitates pilocarpine-induced seizures in prepubertal rats

Stress during gestation can result in early and long-term developmental aberrations. This study aimed to assess the impact of prenatal restraint or predator stress on pilocarpine-induced epileptic behavior. Pregnant rats were exposed to stressors on gestational days 15, 16, and 17. Restraint stress consisted of daily restraint of the dam. During predator stress, caged rats were exposed to a cat in a cage. On postnatal day 25, male pups were injected with pilocarpine and the behavior of each rat was observed. Prenatal stress led to low birth weight and increased blood corticosterone levels. Both stressors significantly potentiated pilocarpine-induced seizures. Predator-stressed pups exhibited significantly severe tonic-clonic seizures compared with restraint-stressed animals. These data emphasize the impact of prenatal stress on fetal growth, and neural and endocrine function. The results also suggest that psychosocial stressors have a greater impact on neural and endocrine function than physical stressors do.     

Behavioral response of the genetically obese (ob/ob) mouse to heat stress: effects of naloxone and prior exposure to immobilization stress

Genetically obese (ob/ob) mice, which possess abnormally elevated levels of pituitary β-endorphin and adrenocorticotropin exhibited less grooming, rearing and jumping during a five min exposure to different levels of heat stress compared to their lean littermate controls (ob/?). Naloxone had a diametrically opposite effect on rearing in these animals, particularly when exposed to low heat stress; it enhanced rearing in ob/ob mice and suppressed the rearing response in ob/? mice. Naloxone enhanced jumping in both the ob/ob mice and the ob/? mice. This effect was slightly, although not significantly, stronger in the obese mice. Finally, exposure to 10 min of immobilization stress before testing at 46°C, enhanced grooming and suppressed jumping in ob/ob and ob/? mice. Naloxone pretreatment reversed the effect of immobilization stress in ob/ob mice but not in their lean littermate controls. The data is discussed in terms of the differential involvement of pituitary endorphins in the behavioral response of ob/ob and ob/? mice to stress.     

Chronic mild restraint stress rats decreased CMKLR1 expression in distinct brain region

Inflammation contributes to the pathophysiology of depression. Chemokine-like receptor-1 (CMKLR1) plays an important role both in the development of inflammation and in the mechanism of antidepressant effect of Omega-3 polyunsaturated fatty acids (Omega-3 PUFAs), ecosapeatanolicacid (EPA). The present study was to investigate the modification of CMKLR1 in chronic restraint stress (CRS) rats. CMKLR1 was examined in different brain region from CRS rats by using western blot and quantitative real-time PCR. The CMKLR1 expression in the hippocampus, prefrontal cortex and cerebellum was determined on 3, 7, 10 and 21 days of repeated restraint stress and was compared to controls. The results showed that the protein and mRNA level of CMKLR1 in the prefrontal cortex and hippocampus were significantly increased on day 3 and then decreased on day 7, 10 and 21 in the CRS rats. The protein and mRNA level of CMKLR1 in cerebellum was similar to that of control group throughout the whole experiment. Changed expression of brain CMKLR1 is suggested to be involved in the mechanism of depression.     

Chronic exposure to stress predisposes to higher autoimmune susceptibility in C57BL/6 mice: Glucocorticoids as a double‐edged sword

Stress activates the hypothalamic‐pituitary‐adrenocortical axis to promote the release of corticosterone (CORT), which consequently suppresses pathogenic stimulation of the immune system. Paradoxically, however, stress often promotes autoimmunity through yet unknown mechanisms. Here we investigated how chronic variable stress (CVS), and the associated alterations in CORT levels, affect the susceptibility to experimental autoimmune encephalomyelitis (EAE) in female and male C57BL/6 mice. Under baseline (nonstressed) conditions, females exhibited substantially higher CORT levels and an attenuated EAE with less mortality than males. However, CVS induced a significantly worsened EAE in females, which was prevented if CORT signaling was blocked. In addition, females under CVS conditions showed a shift toward proinflammatory Th1/Th17 versus Th2 responses and a decreased proportion of CD4⁺CD25⁺ Treg cells. This demonstrates that whereas C57BL/6 female mice generally exhibit higher CORT levels and an attenuated form of EAE than males, they become less responsive to the immunosuppressive effects of CORT under chronic stress and thereby prone to a higher risk of destructive autoimmunity.     

Immobilization and cold stress affect sympatho-adrenomedullary system and pituitary-adrenocortical axis of rats exposed to long-term isolation and crowding

Changes in plasma levels of noradrenaline (NA), adrenaline (A), adrenocorticotropic hormone (ACTH) and corticosterone (CORT), as well as in cytosol glucocorticoid receptor (GR) and heat shock protein 70 (Hsp 70) in hippocampus of adult rat males exposed to two long-term types of psychosocial stress, both under basal conditions and in response to immobilization and cold as heterotypic additional stressor were studied. Long-term isolation produced a significant elevation of basal plasma ACTH and CORT levels, but did not affect that of NA and A, while long-term crowding conditions did not elevate the basal plasma levels of these hormones. Long-term isolation of rats exposed to 2 h of immobilization or cold led to a significant elevation of plasma NA, A and CORT in comparison with the controls. Long-term crowding conditions and exposure of animals to immobilization or cold also resulted in an increased plasma NA, A and CORT levels, but to a lesser extent in comparison with the long-term isolation. At the same time, plasma ACTH was significantly more elevated in long-term crowded than in long-term isolated rats. Both kinds of long-term psychosocial stresses (isolation and crowding) had similar but less pronounced effects on cytosol GR and Hsp 70 concentrations in hippocampus comparing to acute immobilization and cold stress. It seems that long-term psychosocial stresses attenuate the effects of an additional stress on hippocampal GR and Hsp 70 concentrations. These data suggest that individual housing of rats appear to act as a stronger stressor than crowding conditions. When the animals suffering a long-term isolation were exposed to either acute immobilization or cold, a stronger activation of the sympatho-adrenomedullary system (SAS) was recorded in comparison with that found in the long-term crowded group subjected to short-term immobilization or cold. No significant differences in the activity of hypotalamo-pituitary-adrenal (HPA) axis were observed between long-term isolated and long-term crowded rats.     

The deacetylase HDAC6 is a novel critical component of stress granules involved in the stress response

An essential part of the cellular response to environmental stress is a reversible translational suppression, taking place in dynamic cytoplasmic structures called stress granules (SGs). We discovered that HDAC6, a cytoplasmic deacetylase that acts on tubulin and HSP90 and also binds ubiquitinated proteins with high affinity, is a novel critical SG component. We found that HDAC6 interacts with another SG protein, G3BP (Ras-GTPase-activating protein SH3 domain-binding protein 1), and localizes to SGs under all stress conditions tested. We show that pharmacological inhibition or genetic ablation of HDAC6 abolishes SG formation. Intriguingly, we found that the ubiquitin-binding domain of HDAC6 is essential and that SGs are strongly positive for ubiquitin. Moreover, disruption of microtubule arrays or impairment of motor proteins also prevents formation of SGs. These findings identify HDAC6 as a central component of the stress response, and suggest that it coordinates the formation of SGs by mediating the motor-protein-driven movement of individual SG components along microtubules.     

Lager yeasts possess dynamic genomes that undergo rearrangements and gene amplification in response to stress

A long-term goal of the brewing industry is to identify yeast strains with increased tolerance to the stresses experienced during the brewing process. We have characterised the genomes of a number of stress-tolerant mutants, derived from the lager yeast strain CMBS-33, that were selected for tolerance to high temperatures and to growth in high specific gravity wort. Our results indicate that the heat-tolerant strains have undergone a number of gross chromosomal rearrangements when compared to the parental strain. To determine if such rearrangements can spontaneously arise in response to exposure to stress conditions experienced during the brewing process, we examined the chromosome integrity of both the stress-tolerant strains and their parent during a single round of fermentation under a variety of environmental stresses. Our results show that the lager yeast genome shows tremendous plasticity during fermentation, especially when fermentations are carried out in high specific gravity wort and at higher than normal temperatures. Many localised regions of gene amplification were observed especially at the telomeres and at the rRNA gene locus on chromosome XII, and general chromosomal instability was evident. However, gross chromosomal rearrangements were not detected, indicating that continued selection in the stress conditions are required to obtain clonal isolates with stable rearrangements. Taken together, the data suggest that lager yeasts display a high degree of genomic plasticity and undergo genomic changes in response to environmental stress. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Tharappel C. James and Jane Usher contributed equally to this work.