Effects Of Chronic Immobilization Stress Research Articles

Regionalized analysis of gut barrier components in the small intestine of BALB/c mice following chronic immobilization stress.

Microbiota and tight junction proteins (TJPs) are components of the gut barrier, and are considered stress targets that have deleterious effects on intestinal homeostasis. This study aimed to evaluate the effects of chronic immobilization stress on selected small intestine homeostasis parameters. Female BALB/c mice were divided into a stress group that underwent short-term immobilization for 2 h per day for 4 consecutive days, and a non-stressed control group (n = 6 per group). Proximal and distal small intestine samples were excised to assess colony-forming units per gram (CFU/g) of total bifidobacteria in selective agar plates, luminal albumin was assessed using immune-enzymatic assay, pro-inflammatory cytokines were evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and TJPs (pore-forming, claudin (Cld)-2; pore-sealing, Cld-4; ambiguous, Cld-7, -12 and -15) were assessed with RT-qPCR and western blotting. Compared with the control group, the stress group had lower body weight and energy intake. In the distal region, the stressed mice had lower bifidobacteria count and messenger ribonucleic acid (mRNA) expression of Cld-2, Cld-4 and Cld-12, though they had more albumin and higher interleukin (IL)-6 mRNA expression. Within the proximal region, the stressed mice had higher mRNA expression of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), IL-6, Cld-7, Cld-12, and Cld-15, along with lower levels of IL-10 and Cld-4. However, mRNA and protein expression of TJPs were discordant. These findings indicate divergent stress-induced outcomes in the small intestine, evidenced by the elicitation of a pro-inflammatory response and decreased anti-inflammatory response in the duodenum, and by increased albumin transudation and decreased bifidobacterial growth in the distal region.

Effects of chronic immobilization stress on biokinetics and dosimetry of 67Ga in a murine model.

The aim of this work is to determine the effect of chronic immobilization stress on kinetics and dosimetry of 67Ga in a mouse model. A control group (CG) and a stress group (SG), each with 15 mice, were included in the study, and the latter group was subjected to a chronic immobilization stress model 2h daily for 14 consecutive days. At day 13, 67Ga-citrate was administered intraperitoneally (11.24 ± 0.44MBq) to each mouse. Then, sets of three mice were obtained sequentially at 24, 36, 48, 60 and 72h, in which the radionuclide activity was measured with an activity counter. The 67Ga biokinetic data showed a fast blood clearance in the SG, with a mean residence time of 0.06h. The calculated mean radiation absorbed doses were: liver (2.45 × 10-03 Gy), heart (3.17 × 10-04 Gy) and kidney (1.88 × 10-04 Gy) in the SG. The results show that stress reduced weight gain by approximately 13% and also increased adrenal gland weight by 26%. On the other hand, chronic stress accelerates 67Ga clearance after 24h compared to normal conditions. It is concluded that murine organisms under chronic immobilization stress have higher gallium-67 clearance rates, decreasing the cumulated activity and absorbed dose in all organs.

Impact of chronic immobilization stress on parameters of colonic homeostasis in BALB/c mice.

The intestinal epithelium is a monolayer of cells arranged side-by-side and connected by tight junction (TJ) proteins expressed at the apical extreme of the paracellular membrane. This layer prevents stress-induced inflammatory responses, thus helping to maintain gut barrier function and gut homeostasis. The aim of the present study was to evaluate the effects of chronic immobilization stress on the colonic expression of various parameters of homeostasis. A total of two groups of female BALB/c mice (n=6) were included: A stressed group (short-term immobilization for 2 h/day for 4 consecutive days) and an unstressed (control) group. Colon samples were obtained to detect neutrophils and goblet cells by optical microscopy, TJ protein expression (occludin, and claudin −2, −4, −7, −12 and −15) by western blotting, mRNA levels of TJ genes and proinflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, −6 and −8] by reverse transcription-quantitative PCR, fecal lactoferrin by ELISA and the number of colony-forming units of aerobic bacteria. Compared with goblet cells in control mice, goblet cells were enlarged and reduced in number in stressed mice, whereas neutrophil cellularity was unaltered. Stressed mice exhibited reduced mRNA expression for all evaluated TJ mRNAs, with the exception of claudin-7, which was upregulated. Protein levels of occludin and all claudins (with the exception of claudin-12) were decreased in stressed mice. Fecal lactoferrin, proinflammatory cytokine mRNA levels and aerobic bacterial counts were all increased in the stressed group. These results indicated that immobilization stress induced proinflammatory and potential remodeling effects in the colon by decreasing TJ protein expression. The present study may be a useful reference for therapies aiming to regulate the effects of stress on intestinal inflammatory dysfunction.

Glutamine Supplementation Ameliorates Chronic Stress-induced Reductions in Glutamate and Glutamine Transporters in the Mouse Prefrontal Cortex.

Chronic immobilization stress (CIS) induces low levels of glutamate (Glu) and glutamine (Gln) and hypoactive glutamatergic signaling in the mouse prefrontal cortex (PFC), which is closely related to the Glu-Gln cycle. A Gln-supplemented diet ameliorates CIS-induced deleterious changes. Here, we investigated the effects of CIS and Gln supplementation on Glu-Gln cycle-related proteins to characterize the underlying mechanisms. Using the CIS-induced depression mouse model, we examined the expression of 11 proteins involved in the Glu-Gln cycle in the PFC. CIS decreased levels of glutamate transporter 1 (GLT1) and sodium-coupled neutral amino acid transporter (SNAT) 1, SANT2, SNAT3, and SNAT5. Gln supplementation did not affect the non-stressed group but significantly increased GLT1 and SNATs of the stressed group. By immunohistochemical analysis, we confirmed that SNAT1 and SNAT2 were decreased in neurons and GLT1, SNAT3, and SNAT5 were decreased in astrocytes in the medial PFC of the stressed group, but Gln-supplemented diet ameliorated these decrements. Collectively, these results suggest that CIS may cause depressive-like behaviors by decreasing Glu and Gln transportation in the PFC and that a Gln-supplemented diet could prevent the deleterious effects of CIS.

Inactivation of basolateral amygdala prevents chronic immobilization stress-induced memory impairment and associated changes in corticosterone levels

Chronic stress causes detrimental effects on various forms of learning and memory. The basolateral amygdala (BLA) not only plays a crucial role in mediating certain forms of memory, but also in the modulation of the effects of stress. Chronic immobilization stress (CIS) results in hypertrophy of the BLA, which is believed to be one of the underlying causes for stress’ effects on learning. Thus, it is plausible that preventing the effects of CIS on amygdala would preclude its deleterious cognitive effects. Accordingly, in the first part, we evaluated the effect of excitotoxic lesion of the BLA on chronic stress-induced hippocampal-dependent spatial learning using a partially baited radial arm maze task. The BLA was ablated bilaterally using ibotenic acid prior to CIS. Chronically stressed rats showed impairment in spatial learning with decreased percentage correct choice and increased reference memory errors. Excitotoxic lesion of the BLA prevented the impairment in spatial learning and reference memory. In the retention test, lesion of the BLA was able to rescue the chronic stress-induced impairment. Interestingly, stress-induced enhanced plasma corticosterone levels were partially prevented by the lesion of BLA. These results motivated us to evaluate if the same effects can be observed with temporary inactivation of BLA, only during stress. We found that chronic stress-induced spatial learning deficits were also prevented by temporary inactivation of the BLA. Additionally, temporary inactivation of BLA partially precluded the stress-induced increase in plasma corticosterone levels. Thus, inactivation of BLA precludes stress-induced spatial learning deficits, and enhanced plasma corticosterone levels. It is speculated that BLA inactivation-induced reduction in corticosterone levels during stress, might be crucial in restoring spatial learning impairments. Our study provides evidence that amygdalar modulation during stress might be beneficial for strategic management of stress-related cognitive deficits.

The Effects of Immobilization Stress on Serum Ghrelin Level, Food Intake and Body Weight in Male and Female Rats

Background: Stress causes various physiological, histological and behavioral changes. Objectives: The goal of the present study was to evaluate the effects of chronic immobilization stress on plasma ghrelin level, food intake and body weight in both male and female rats. Materials and Methods: Twenty male and 20 female rats were studied. These animals were divided to four groups (two control groups and two experimental groups). In the experimental groups, rats were exposed to immobilization stress for 14 days. On the fourteenth day serum ghrelin level was measured by the ELISA kit. Food consumption and body weight change during 14 days were also recorded. Student t-test was used to compare mean among the groups. Results: In male rats, immobilization stress significantly (P ˂ 0.0001) increased serum ghrelin level and decreased body weight as compared with the control group (P ˂ 0.0001). Food consumption significantly (P ˂ 0.0001) increased in stressed female and male rats. Conclusions: The results of this study indicate that the effect of stress on serum ghrelin level and body weight is gender-depended.

Protective Effects of Resveratrol against Chronic Immobilization Stress on Testis

Objective. The aim of this study was to investigate protective effects of resveratrol, a strong antioxidant, against possible negative effects of chronic immobilization stress on testes of male rats histochemically, immunohistochemically, ultrastructurally, and biochemically. Material and Methods. Male Wistar rats were divided into 4 groups (n = 7). Group I, control group (C), was not exposed to stress. Group II, stress group (S), was exposed to chronic immobilization stress. In Group III, low dose resveratrol + stress group (LRS), rats were given 10 mg/kg/day resveratrol just before the stress application. In Group IV, high dose resveratrol + stress group (HRS), rats were given 20 mg/kg/day resveratrol just before the stress application. For chronic immobilization stress application animals were put in the plastic tubes (6 cm in diameter, 15 cm in length) during 32 days for 6 hours. All animals were sacrificed 18 hours after the last stress application. Results. Histochemical and ultrastructural investigations showed that in stress group there was germ cell deprivation in seminiferous tubules and increase of connective tissue on interstitial area. No significant changes were seen in low and high dose resveratrol groups. After immunohistochemical investigations, TUNEL (+) and Active Caspase-3 (+) cells were increased in seminiferous tubules of stress group compared with those control group, but they were decreased in low and high dose resveratrol groups. According to biochemically results, MDA, GSH, and testosterone levels in stress group showed no significant difference when compared with those of the other groups. Conclusion. The chronic immobilization stress increases oxidative stress and apoptosis and causes histological tissue damages; resveratrol can minimize the histological damage in testes significantly.

Depletion of polysialic acid from neural cell adhesion molecule (PSA-NCAM) increases CA3 dendritic arborization and increases vulnerability to excitotoxicity

Chronic immobilization stress (CIS) shortens apical dendritic trees of CA3 pyramidal neurons in the hippocampus of the male rat, and dendritic length may be a determinant of vulnerability to stress. Expression of the polysialylated form of neural cell adhesion molecule (PSA-NCAM) in the hippocampal formation is increased by stress, while PSA removal by Endo-neuraminidase-N (endo-N) is known to cause the mossy fibers to defasciculate and synapse ectopically in their CA3 target area. We show here that enzymatic removal of PSA produced a remarkable expansion of dendritic arbors of CA3 pyramidal neurons, with a lesser effect in CA1. This expansion eclipsed the CIS-induced shortening of CA3 dendrites, with the expanded dendrites of both no-stress-endo-N and CIS-endo-N rats being longer than those in no-stress-control rats and much longer than those in CIS-control rats. As predicted by the hypothesis that endo-N-induced dendritic expansion might increase vulnerability to excitotoxic challenge, systemic injection with kainic acid, showed markedly increased neuronal degeneration, as assessed by fluorojade B histochemistry, in rats that had been treated with endo-N compared to vehicle-treated rats throughout the entire hippocampal formation. PSA removal also exacerbated the CIS-induced reduction in body weight and abolished effects of CIS on NPY and NR2B mRNA levels. These findings support the hypothesis that CA3 arbor plasticity plays a protective role during prolonged stress and clarify the role of PSA-NCAM in stress-induced dendritic plasticity.

Effects of chronic immobilization stress on anxiety-like behavior and basolateral amygdala morphology in Fmr1 knockout mice

Several lines of clinical evidence support the idea that fragile X syndrome (FXS) may involve a dysregulation of hypothalamic-pituitary-adrenal axis function [Wisbeck et al. (2000) J Dev Behav Pediatr 21:278-282; Hessl et al. (2002) Psychoneuroendocrinology 27:855-872]. We had tested this idea in a mouse model of FXS (Fmr1 KO) and found that the hormonal response to acute stress was similar to that of wild-type (WT) mice [Qin and Smith (2008) Psychoneuroendocrinology 33:883-889]. We report here responses to chronic stress (CS) in Fmr1 KO mice. Following restraint for 120 min/d, 10 consecutive days, we assessed dendrite and spine morphology in basolateral amygdala (BLA). We also monitored behavior in an elevated plus maze (EPM) and the hormonal response to this novel spatial environment. After CS, mice of both genotypes underwent adrenal hypertrophy, but effects were greater in WT mice. Behavior in the EPM indicated that only WT mice had the expected increase in anxiety following CS. Serum corticosterone and adrenocorticotropic hormone (ACTH) levels were both increased following the spatial novelty of EPM, and there were no differences between genotypes in the hormonal responses. BLA dendritic branching increased proximal to the soma in WT, but in Fmr1 KO mice branching was unaffected close to the soma and slightly decreased at one point distal to the soma. Similarly, spine density on apical and basal dendrites increased in WT but decreased in Fmr1 KO mice. Spine length on apical and basal dendrites increased in WT but was unaffected in Fmr1 KO mice. These differences in behavioral response and effects on neuron morphology in BLA suggest a diminished adaptive response of Fmr1 KO mice.

Chronic stress-induced hippocampal dendritic retraction requires CA3 NMDA receptors

Chronic stress induces dendritic retraction in the hippocampal CA3 subregion, but the mechanisms responsible for this retraction and its impact on neural circuitry are not well understood. To determine the role of NMDA (N-methyl- d-aspartic acid) receptor (NMDAR)-mediated signaling in this process, we compared the effects of chronic immobilization stress (CIS) on hippocampal dendritic morphology, hypothalamic-pituitary-adrenal (HPA) axis activation, and anxiety-related and hippocampus-dependent behaviors, in transgenic male mice in which the NMDAR had been selectively deleted in CA3 pyramidal cells and in non-mutant littermates. We found that CIS exposure for 10 consecutive days in non-mutant mice effectively induces HPA axis activation and dendritic retraction of CA3 short-shaft pyramidal neurons, but not CA3 long-shaft pyramidal neurons, suggesting a differential cellular stress response in this region. Dendritic reorganization of short-shaft neurons occurred throughout the longitudinal axis of the hippocampus and, in particular, in the ventral pole of this structure. We also observed a robust retraction of dendrites in dorsal CA1 pyramidal neurons in the non-mutant C57BL/6 mouse strain. Strikingly, chronic stress-induced dendritic retraction was not evident in any of the neurons in either CA3 or CA1 in the mutant mice that had a functional lack of NMDARs restricted to CA3 pyramidal neurons. Interestingly, the prevention of dendritic retraction in the mutant mice had a minimal effect on HPA axis activation and behavioral alterations that were induced by chronic stress. These data support a role for NMDAR-dependent glutamatergic signaling in CA3 in the cell-type specific induction of dendritic retraction in two hippocampal subregions following chronic stress.

Effects of Chronic Immobilization Stress on the Expression of Fas/FasL in the Brain of Rats

Currently,there exists a substantial body of evidence suggesting that chronic stress can increase susceptibility to diseases such as depression and posttraumatic stress disorder.Chronic stress induces neuronal atrophy and death in the cortex as well as in the hippocampus.Proto-oncogene protein kinase B(PKB),also known as Akt,plays a central role in a signaling pathway whose various components have been linked to cellular survival.In recent years,Akt as well as one of its downstream targets Forkhead transcription factors(forkhead homologue in rhabdomyosarcoma,(FKHR)) and the Fas/FasL system have emerged as cardinal pathways underlying cellular survival and opposing apoptosis in neurons.In our previous experiments,we observed that repeated immobilization stress exposure can change the levels of phosphorylation of Akt and FKHRL1 in the rat hippocampus.However,the activation of the Fas/FasL system following chronic stress in the brain remains poorly defined.We hypothesized that repeated immobilization stress exposure may change the levels of the Fas/FasL system in the brains of rats.The purpose of the present study was to detect the effects of chronic immobilization stress on the expression of Fas/FasL in the prefrontal cortex,entorhinal cortex,and hippocampus in rats. Twenty four male Sprague-Dawley rats(280±20g) were purchased from Shanghai Laboratory Animal Center,Chinese Academy Sciences,and they were used for all experiments.These rats were randomly divided into three groups-the immobilization stressed group,apparatus control group,and control group.Each group comprised eight rats.The rats were housed in groups of four,under standard laboratory conditions in temperature-controlled rooms(24℃),and maintained under a 12-h light/dark cycle(lights on at 0800 hours) with food pellets and water available ad libitum.Rats from the immobilization stressed group were immobilized by fixing a board on their backs for an hour each day for 14 consecutive days.The apparatus controls were placed in a novel environment for an hour per day for 14 consecutive days.The controls were free of stress.At the end of the experimental period,the rats were decapitated and their prefrontal cortex,entorhinal cortex,and hippocampi were rapidly removed and maintained at-80℃ until analysis.The protein levels of Fas and FasL were determined by Western blotting.The intensities of the bands corresponding to the protein of interest were quantified using scanning densitometry and compared using one-way ANOVA.The adrenal gland and thymus gland indices were also calculated on the day following the 14-day-stress modeling.Organ index = wet weight/weight of rat×100.The statistical significance was determined at p 0.05. A total of 24 rats were considered in the analysis of the results.The body weight gain of the stressed group was significantly lower than that of the control group and the apparatus control group.A significant change was observed in the organ indices of the stressed groups as compared to the control group.The Fas and FasL levels of the hippocampus revealed a significant difference among three groups(F= 26.9,p 0.001;F= 40.29,p 0.01,respectively).The Fas and FasL levels were significantly increased in the immobilization stressed group than in the control group(p 0.01).The Fas levels of the prefrontal cortex and entorhinal cortex in the immobilization stressed group were significantly increased than those in the control group(p 0.01).In contrast,the FasL levels of the prefrontal cortex and entorhinal cortex did not reveal a significant deference between the immobilization stressed group and control group. The results suggested that chronic immobilization stress can induce a more significant increase in both the Fas and FasL levels in the hippocampus.Changing the expression of Fas/FasL may be an effective biological predictor for the change of structure or function of hippocampus induced by chronic stress in rats.

Chronic immobilization stress alters aspects of emotionality and associative learning in the rat.

Chronic stress significantly alters limbic neuroarchitecture and function, and potentiates emotionality in rats. Chronic restraint stress (CRS) increases aggression among familiar rats, potentiates anxiety, and enhances fear conditioning. Chronic immobilization stress (CIS) induces anxiety behavior and dendritic hypertrophy in the basolateral amygdala, which persist beyond a recovery period. However, little else is known about the emotional impact of CIS as a model of chronic stress or depression. Therefore, the authors present two experiments examining emotional and learned responses to CIS. In Experiment I, the authors examine individual differences in behaviors during and after CIS, specifically: struggling, aggression, learned helplessness, inhibitory avoidance, and escape behavior. In Experiment II, the authors confirm the effects of CIS on aggression and struggling during immobilization, and correlate individual responses with aspects of conditioned fear. Here the authors report significant effects of CIS on aggression, inhibitory avoidance, escape, as well as learned aspects of fear (i.e., fear conditioning) and inescapable stress (i.e., struggling and helplessness). These results emphasize the emotional and learned responses to CIS evident during and after the stress treatment, as well as the importance of individual differences.

Changes of Glucocorticoid Receptor and Levels of CRF mRNA, POMC mRNA in Brain of Chronic Immobilization Stress Rats

The effect of chronic immobilization stress (CIS) on the biochemical parameters has been one of the hot topics in neuroscience. The aim of this study was to investigate the effects of CIS on the levels of glucocorticoid receptor (GR), corticotrophin-releasing factor (CRF) mRNA and proopiomelanocortin (POMC) mRNA in brains of rats. The rats were randomly divided into stressed and control groups. The stressed group was given CIS 3 h a day for 21 days continuously. GR of rats' hippocampus and prefrontal cortex (PFC) were detected by immunohistochemistry method. In addition, the CRF mRNA and POMC mRNA of rats' brain regions (hypothalamus, pituitary, hippocampus, and PFC) were detected by reverse transcription-polymerase chain reaction (RT-PCR). After exposure to CIS for 21 days, the GR immuno staining (the gray values) of the stressed group was less than that of the control group in hippocampal CA(1), dentate gyrus, and PFC (P < 0.01). Quantitative analysis indicated the presence of CRF mRNA in hypothalamus and pituitary, while POMC mRNA in PFC, hippocampus and pituitary of the stressed group was less than that of the control group (P < 0.01). The decreased levels of GR, CRF mRNA, and POMC mRNA in different brain regions may contribute to explanation of the CIS induced mechanism.

Chronic-stress induced modulation of different states of anxiety-like behavior in female rats

Stress facilitates emotionality and consolidation of aversive memories in male rodents. In addition, considerable sexual dimorphism has been observed in animal and clinical literature, both in response to stress and predisposition to anxiety disorders thought to be exacerbated by stress. In view of this, we investigated effects of chronic immobilization stress and chronic unpredictable stress on anxiety-like behavior exhibited by female Wistar rats, using the elevated plus-maze. Neither of the stress paradigms employed in this study significantly influenced anxiety, as manifested by similar open-arm exploration in control and treated animals. Previous studies have reported that in males, exposure to elevated plus-maze during an initial trial significantly reduces open-arm exploration in subsequent retesting, an effect attributed to consolidation of aversive experience of the initial exposure. Control female animals, during a second exposure to the maze 72 h after the first trial, displayed a similar shift to a state of enhanced anxiety. Furthermore, exposure to stress did not affect such consolidation of anxiety, as evidenced by similar reduction in open-arm exploration between control and stressed animals during retesting. We conclude that female rats are insensitive to chronic stress in terms of facilitation and consolidation of anxiety.

Effects of chronic stress on dendritic arborization in the central and extended amygdala

A differential role has been suggested for two important areas in the neural circuitry of stress, central nucleus of the amygdala (CeA) and bed nucleus of stria terminalis (BNST) in the extended amygdala, in regulating fear versus anxiety. Since chronic stress enhances anxiety and consolidation of aversive memories, we examined the effects of chronic immobilization stress (CIS) on neuronal morphology in the CeA and BNST of rats. In contrast to previous reports of stress-induced atrophy in the hippocampus, CIS does not cause dendritic atrophy in CeA and BNST neurons. While dendritic arborization in CeA neurons remains unaffected, it increases in BNST neurons after CIS. These results suggest a role for dendritic remodeling of BNST neurons in stress-induced facilitation of anxiety.

The Ca2+ Channel Blockade Changes the Behavioral and Biochemical Effects of Immobilization Stress

We investigated how the effects of chronic immobilization stress in rats are modified by Ca2+ channel blockade preceding restraint sessions. The application of nifedipine (5 mg/kg) shortly before each of seven daily 2 h restraint sessions prevented the development of sensitized response to amphetamine as well as the stress-induced elevation of the densities of L-type Ca2+ channels in the hippocampus and significantly reduced the elevation of the densities of [3H]nitrendipine binding sites in the cortex and D1 dopamine receptors in the limbic forebrain. Neither stress, nor nifedipine affected the density of α1-adrenoceptors and D1 receptors in the cerebral cortex nor D2 dopamine receptors in the striatum. A single restraint session caused an elevation of blood corticosterone level that remained unaffected by nifedipine pretreatment, but the reduction of this response during the eighth session was significantly less expressed in nifedipine-treated rats. We conclude that L-type calcium channel blockade prevents development of several stress-induced adaptive responses.

Effects of chronic immobilization stress on GH and TSH secretion in the rat: Response to hypothalamic regulatory factors

The effect of chronic immobilization (2 h/day) for 13 days on basal and stress levels of GH and TSH, and their response to various hypothalamic regulatory factors was studied in male Sprague-Dawley rats. Chronic immobilization (IMO) resulted in reduced serum TSH levels in stress situations but not in resting conditions. GH secretion was inhibited both in resting and stress situations. Chronic IMO impaired both GH and TSH responses to GRH and TRH, respectively, but also to another peptide (VIP) stimulatory for the two hormones. Whereas somatostatin administration inhibited GH secretion in control but not in chronic IMO rats, its inhibitory effect on TSH was slight and similar in the two experimental groups. The present results suggest that chronic exposure to a severe stressor such as IMO alters GH and TSH secretion, at least in part by changes in the response of the pituitary to the hypothalamic regulatory factors. The actual influence of chronic IMO on the release of these peptides into the median eminence remains to be studied.

Estrogen influences the effect of immobilization stress on immunoreactive beta-endorphin levels in the female rat pituitary.

Immunoreactive beta-endorphin (IR-BE) levels in the plasma, anterior pituitary (AP), the neurointermediate lobe of the pituitary (NIL), and the hypothalamus were determined in castrated female rats and castrated female rats treated with estradiol benzoate (estrogen), after exposure to acute (once for 45 min) or chronic (45 min each day for 15 consecutive days) immobilization stress. Acute and chronic stress increased plasma levels of IR-BE to the same extent in castrated female rats and castrated female rats treated with estrogen. In castrated female rats, acute stress produced an increase in the concentration of IR-BE in the AP, which was attenuated by the administration of estrogen. Although IR-BE in the NIL was not influenced by acute stress in castrated animals, exposure to acute stress resulted in an elevation in IR-BE levels in the NIL of rats given estrogen. Chronic stress did not affect the concentration of IR-BE in the AP of castrated females or castrated females treated with estrogen. Chronic stress did, however, increase the concentration of IR-BE in the NIL of castrated animals. This affect of stress on IR-BE levels in the NIL was potentiated by estrogen administration. IR-BE levels in the hypothalamus were reduced by estrogen and were not affected by acute or chronic stress, regardless of the gonadal steroid environment. As determined by column chromatography, administration of estrogen, as well as subjection to chronic stress, promoted the processing of the proopiomelanocortin precursor to form beta-lipotropin rather than beta-endorphin in the AP. By these methods, the only immunoreactivity detected in the NIL and the hypothalamus was beta-endorphin. These data indicate that IR-BE levels in the plasma, the AP, and the NIL of female rats are affected by immobilization stress and that estrogen modulates the effects of acute immobilization stress on IR-BE levels in the AP and the NIL and the effects of chronic immobilization stress on the levels of IR-BE in the NIL.

Effects of chronic immobilization stress on pituitary hormone secretion, on hypothalamic factor levels, and on pituitary responsiveness to LHRH and TRH in female rats.

The effects of 8 h of daily immobilization applied for 10 consecutive days on pituitary and plasma levels of anterior pituitary (AP) hormones as well as on TRH, LHRH and somatostatin (SRIF) content in