Days Of Chronic Stress Research Articles

Octopamine mediates sugar relief from a chronic-stress-induced depression-like state in Drosophila

Chronic, uncontrollable stress can result in psychiatric syndromes, including anxiety and major depressive disorder, in humans and mammalian disease models.1,2 Similarly, several days of chronic stress can induce depression-associated behavioral alteration in Drosophila accompanied by changes in biogenic amine levels in the adult brain.3-6 In our chronic stress paradigm, flies are subjected to 3days of repetitive phases of 300Hz vibrations combined with overcrowding and food deprivation. This treatment reduces voluntary behavioral activity, including the motivation to climb wide gaps (risk taking) and to stop for sweets (anhedonia), suggesting a depression-like state (DLS). These behavioral changes correlate with decreased serotonin release to the mushroom body (MB), a major behavioral control center in the central brain of the fly.7,8 Stressed flies are relieved from the DLS by feeding the anti-depressant serotonin precursor 5-HTP or the selective serotonin reuptake inhibitor fluoxetine. Notably, feeding sucrose to stressed flies results in elevated serotonin levels in the brain and ameliorates the DLS.3 Here, we show that this sugar relief is mediated by the neurotransmitter octopamine signaled from ventral unpaired medial neurons located in the subesophageal ganglion. The octopamine signaling of sweet sensation is transmitted to the MB via the dopaminergic PAM neurons. In addition, neuronal-silencing experiments reveal that the serotonergic dorsal paired medial (DPM) neurons innervating the MB are essential for sugar relief. Conversely, thermogenetic or optogenetic activation of DPMs can replace sweet sensation, elucidating that serotonergic signaling from DPMs takes part in positively modulating DLS-related behavioral changes.

ELECTRONIC-MICROSCOPIC CHANGES OF THE INTRAMURAL NERVOUS PLEXUS OF THE DUODENUM IN CONDITIONS OF CHRONIC STRESS

The article presents a study of ultrastructural changes of neurons of the intramural nervous system (muscular- intestinal plexus) of the duodenum under chronic stress. The experiment was performed on 60 white adult male rats. The material was taken from the proximal and distal parts of the duodenum after 14, 28, 56 days of the experiment. For electron microscopic examination, pieces of the duodenum were fi xed in 2.5% glutaraldehyde solution for 4 hours, and then fi xed in 2% osmium tetroxide solution. First, semi-thin sections were made to detect the localization of nerve cells of the musculoskeletal plexus of the duodenum, staining them in 1% methylene blue solution or 0.1% toluidine blue solution. Ultrathin sections were made on an ultramicrotome Tesla BS-490A, the material was studied on an electron microscope TEM – 125 K («Selmi», Ukraine). After 14 days of chronic stress, there is signifi cant damage to the neurons of the musculoskeletal plexus of the duodenum due to reactive and edematous processes, accompanied by the development of ultrastructural changes and the death of some neurons. After 28-56 days of the experiment, edematous, dystrophic- destructive processes in nerve cells are further observed, but in some of them there are a number of compensatory- adaptive and reparative processes aimed at restoring impaired functions.

Environmental enrichment decreases chronic psychosocial stress-impaired extinction and reinstatement of ethanol conditioned place preference in C57BL/6 male mice.

During the last few decades, alcohol use disorders (AUD) have reached an epidemic prevalence, yet social influences on alcoholism have not been fully addressed. Several factors can modulate alcohol intake. On one hand, stress can reinforce ethanol-induced behaviors and be an important component in AUD and alcoholism. On the other hand, environmental enrichment (EE) has a neuroprotective role and prevents the development of excessive ethanol intake in rodents. However, studies showing the role of EE in chronic psychosocial stress-impaired ethanol-conditioned rewards are nonexistent. The purpose of the current study is to explore the potential protective role of EE on extinction and reinstatement of ethanol-conditioned place preference (EtOH-CPP) following chronic psychosocial stress. In the first experiment and after the EtOH-CPP test, the mice were subjected to 15days of chronic stress, then housed in a standard (SE) or enriched environment (EE) while EtOH-CPP extinction was achieved by repeated exposure to the CPP chambers without ethanol injection. In the second experiment and after the EtOH-CPP test, extinction was achieved as described above. Mice were then exposed to chronic stress for 2weeks before being housed in a SE or EE. EtOH-CPP reinstatement was induced by a single exposure to the conditioning chambers. As expected, stress exposure increased anxiety-like behavior and reduced weight gain. More importantly, we found that EE significantly shortened chronic stress-delayed extinction and decreased the reinstatement of EtOH-CPP. These results support the hypothesis that EE reduces the impact of alcohol-associated environmental stimuli, and hence it may be a general intervention for reducing cue-elicited craving and relapse in humans.

Gradual decrease in spermatogenesis caused by chronic stress

Chronic stress induces decreased sperm motility, viability and concentration in stressed males. Also, stress modifies oxidative status and causes apoptosis in testes, as well as a decrease in the epithelial area of seminiferous tubules. However, there are no studies that analyze the alterations caused by stress in testicular cells. Thus, in this study, alterations in the morphology of testicular germ cells caused by different days of chronic stress were assessed. Adult male rats were exposed to stress by immersion in cold water (ICW) daily for 3, 8, 20 or 50 consecutive days. Plasma testosterone and corticosterone were also assessed. Results showed that chronic stress causes loss of germ cells, and alteration of spermatogenesis. Seminiferous tubules from stressed males showed several degenerative signs, such as vacuoles in the basal epithelium, with picnotic indicia; moderate to severe exfoliation of degenerative germinal cells in the tubule lumen was also observed. These alterations were observed in all days of stress in a gradual way, from day 3–50. Testosterone levels were decreased at all those times, and corticosterone concentrations were increased on the same days. These results show that chronic stress causes severe damage to germ cells, which can account for infertility problems in males. These alterations are related to a decrease in testosterone as well as an increase in corticosterone caused by stress.

Early hippocampal volume loss as a marker of eventual memory deficits caused by repeated stress.

Exposure to severe and prolonged stress has detrimental effects on the hippocampus. However, relatively little is known about the gradual changes in hippocampal structure, and its behavioral consequences, over the course of repeated stress. Behavioral analyses during 10 days of chronic stress pointed to a delayed decline in spatial memory, the full impact of which is evident only after the end of stress. In contrast, concurrent volumetric measurements in the same animals revealed significant reduction in hippocampal volumes in stressed animals relative to their unstressed counterparts, as early as the third day of stress. Notably, animals that were behaviorally the worst affected at the end of chronic stress suffered the most pronounced early loss in hippocampal volume. Together, these findings support the view that not only is smaller hippocampal volume linked to stress-induced memory deficits, but it may also act as an early risk factor for the eventual development of cognitive impairments seen in stress-related psychiatric disorders.

Central immune alterations in passive strategy following chronic defeat stress

The relationship between stress, mood disorders and immune disorders is known, but what remains to be resolved is why certain individuals are more susceptible than others to suffer different disorders, along with the biological mechanisms that underlie these differences. The objective of this study was to analyze the changes in the expression patterns of proinflammatory cytokines in the hypothalamus, hippocampus, amygdala and prefrontal cortex after chronic defeat, depending on the coping strategy used. The expression levels of α1b and α2a adrenergic receptors and cytokine-inducible nitric oxide synthase (iNOS) in the prefrontal cortex were also measured. The results indicated that subjects with a passive coping strategy showed high levels of interleukin-6 (IL-6) and interleukin-1β (IL-1β) expression in several cerebral structures in resting conditions after 21 days of chronic stress and increases in these cytokine levels in the hippocampus following an additional stress. Low expression levels of tumour necrosis factor-alpha (TNF-α) in the prefrontal cortex in active subjects at rest and in passive subjects after an additional defeat were detected. The iNOS expression levels were lower in the prefrontal cortex of the active group at rest. With respect to adrenergic receptor expression, there were no changes as a function of stress, but there were changes as a function of coping strategy. These results indicate differences in the variables studied in terms of the coping strategy adopted, with passive subjects having a biological profile that could be considered more vulnerable to the development of stress-related disorders.

Oxidative status in testis and epididymal sperm parameters after acute and chronic stress by cold-water immersion in the adult rat

Stress is associated with detrimental effects on male reproductive function. It is known that stress increases reactive oxygen species (ROS) generation in the male reproductive tract. High ROS levels may be linked to low sperm quality and male infertility. However, it is still not clear if ROS are generated by stress in the testis. The objective of this study was to characterize the role of oxidative stress induced by cold-water immersion stress in the testis of adult male rats and its relation with alterations in cauda epididymal sperm. Adult male rats were exposed to acute stress or chronic stress by cold-water immersion. Rats were sacrificed at 0, 6, 12, and 24 hours immediately following acute stress exposure, and after 20, 40, and 50 days of chronic stress. ROS production increased only at 6 hours post-stress, while the activity and expression of antioxidant enzymes, lipid peroxidation (LPO), and sperm parameters were not modified in the testis. Corticosterone increased immediately after acute stress, whereas testosterone was not modified. After chronic stress, testicular absolute weight decreased; in addition, ROS production and LPO increased at 20, 40, and 50 days. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) decreased throughout the duration of chronic stress and the activity of catalase (CAT) decreased at 40 and 50 days, and increased at 20 days. The expression of copper/zinc superoxide dismutase (SOD1) and CAT were not modified, but the expression of phospholipid hydroperoxide glutathione peroxidase (GPx-4) decreased at 20 days. Motility, viability, and sperm count decreased, while abnormal sperm increased with chronic stress. These results suggest that during acute stress there is a redox state regulation in the testis since no deleterious effect was observed. In contrast, equilibrium redox is lost during chronic stress, with low enzyme activity but without modifying their expression. In addition, corticosterone increased while testosterone decreased, this decrease is related to the negative effects seen in sperm.

The dynamic impact of repeated stress on the hippocampal spatial map.

Stress alters the function of many physiological processes throughout the body, including in the brain. A neural circuit particularly vulnerable to the effects of stress is the hippocampus, a key component of the episodic and spatial memory system in both humans and rodents. Earlier studies have provided snapshots of morphological, molecular, physiological and behavioral changes in the hippocampus following either acute or repeated stress. However, the cumulative impact of repeated stress on in vivo hippocampal physiology remains unexplored. Here we report the stress-induced modulation of the spatially receptive fields of the hippocampal CA1 'place cells' as mice explore familiar and novel tracks after 5 and 10 days of immobilization stress. We find that similar to what has been observed following acute stress, five days of repeated stress results in decreased excitability of CA1 pyramidal cells. Following ten days of chronic stress, however, this decreased hippocampal excitability is no longer evident, suggesting adaptation may have occurred. In addition to these changes in neuronal excitability, we find deficient context discrimination, wherein both short-term and chronic stress impair the ability of the hippocampus to unambiguously distinguish novel and familiar environments. These results suggest that a loss of network flexibility may underlie some of the behavioral deficits accompanying chronic stress.

Chronic psychosocial stress causes delayed extinction and exacerbates reinstatement of ethanol-induced conditioned place preference in mice

We have shown previously, using an animal model of voluntary ethanol intake and ethanol-conditioned place preference (EtOH-CPP), that exposure to chronic psychosocial stress induces increased ethanol intake and EtOH-CPP acquisition in mice. Here, we examined the impact of chronic subordinate colony (CSC) exposure on EtOH-CPP extinction, as well as ethanol-induced reinstatement of CPP. Mice were conditioned with saline or 1.5g/kg ethanol and were tested in the EtOH-CPP model. In the first experiment, the mice were subjected to 19days of chronic stress, and EtOH-CPP extinction was assessed during seven daily trials without ethanol injection. In the second experiment and after the EtOH-CPP test, the mice were subjected to 7days of extinction trials before the 19days of chronic stress. Drug-induced EtOH-CPP reinstatement was induced by a priming injection of 0.5g/kg ethanol. Compared to the single-housed colony mice, CSC mice exhibited increased anxiety-like behavior in the elevated plus maze (EPM) and the open field tests. Interestingly, the CSC mice showed delayed EtOH-CPP extinction. More importantly, CSC mice showed increased alcohol-induced reinstatement of the EtOH-CPP behavior. Taken together, this study indicates that chronic psychosocial stress can have long-term effects on EtOH-CPP extinction as well as drug-induced reinstatement behavior and may provide a suitable model to study the latent effects of chronic psychosocial stress on extinction and relapse to drug abuse.

Stress Worsens Endothelial Function and Ischemic Stroke via Glucocorticoids

Chronic stress is associated with increased stroke risk. However, the underlying pathophysiological mechanisms are poorly understood. We examined the effects of chronic stress on endothelial function and ischemic brain injury in a mouse model. 129/SV mice were treated with glucocorticoid receptor antagonist mifepristone (25 mg kg(-1)/d) or vehicle and exposed to 28 days of chronic stress consisting of exposure to rat, restraint stress, and tail suspension. Heart rate and blood pressure were continuously recorded by telemetry. Endothelial nitric oxide synthase mRNA and protein expression as well as superoxide production and lipid hydroperoxides were quantified. Endothelium-dependent vasorelaxation was measured in aortic rings. Ischemic lesion volume was quantified after 30 minutes filamentous middle cerebral artery occlusion and 72 hours reperfusion. Chronic stress caused a significant increase in heart rate, impaired endothelium-dependent vasorelaxation, increased superoxide production, and reduced aortic and brain endothelial nitric oxide synthase levels. Animals exposed to chronic stress showed major increases in ischemic lesion size. These deleterious effects of stress were completely reversed by treatment with mifepristone. Chronic stress increases stroke vulnerability likely through endothelial dysfunction, which can be reversed by a glucocorticoid receptor antagonist.

Combined effects of molt and chronic stress on heart rate, heart rate variability, and glucocorticoid physiology in European Starlings

Molt is an important life-history stage in avian species, but little is known about the effects of chronic stress during this period. Three weeks after the onset of molt, captive European starlings ( Sturnus vulgaris) were exposed to 18 days of chronic stress, induced with four 30-minute randomized stressors presented daily. Birds showed no chronic-stress-induced changes in heart rate or heart rate variability when measured either during the middle of the day or at night. These data suggest that chronic stress did not alter the balance between sympathetic and parasympathetic nervous system regulation of cardiovascular function, which contrasts with data from an earlier study indicating that chronic stress profoundly alters cardiovascular function in non-molting starlings. Additionally, there was a significant increase in restraint-induced corticosterone secretion the first week of chronic stress that subsequently returned to pre-chronic-stress levels by the second week of exposure. The attenuated corticosterone response again contrasts with data from non-molting starlings that showed significant decreases in corticosterone responses. Consequently, the resistance to cardiovascular and corticosterone changes indicates that the physiological changes induced by chronic stress are greatly attenuated in molting birds. Overall, the data suggest that molt requires a degree of physiological stability that must be protected, so that if a bird is exposed to chronic stress during this life-history stage, molt takes priority.

Effects of enrichment predominate over those of chronic stress on fear-related behavior in male rats

The ability to discriminate between spatial contexts is crucial for survival. This ability can be succinctly tested in the paradigm of fear renewal. In this paradigm, a change of spatial context results in robust renewal of conditioned fear, even if the conditioned fear has been previously extinguished. Chronic stress and environmental enrichment are known to affect learning and memory in opposite directions, with the former generally being deleterious. In this study, we examined the effects of chronic stress and enrichment on fear renewal in rats. Fear was evaluated as freezing responses to an auditory conditioning stimulus initially associated with footshocks in context A; fear extinction was evaluated in a novel spatial context (B) without the conditioned stimulus, and renewal in a third context (C) with the auditory cue. Specifically, we aimed to test if environmental enrichment can oppose the effects of chronic stress on fear renewal. We exposed different groups of adult male Wistar rats (6–12 per group) to 10 days of chronic stress (immobilization for 2 h daily), 14 days of enrichment, or a combination of both. We report that chronic stress compromised fear extinction and renewal. In contrast, enrichment re-established fear renewal in chronically stressed rats. Enhanced contextual modulation of fear memories in animals experiencing environmental enrichment while stressed could reflect an adaptive response. This could allow greater flexibility to optimize vigilance in differing spatial contexts.

Sleep pattern in rats under different stress modalities

The present study was designed to evaluate the sleep pattern of rats submitted to chronic stressors (restraint, electrical footshock, swimming and cold) applied to male rats. After 48 h-baseline recording, rats were submitted to 4 days of chronic stress, and electrocorticogram recordings were carried out continuously. The stressors (footshock, swimming and cold) were applied twice a day for periods of 1 h at 9:00 and 16:00 h. Restrained animals were maintained in plastic cylinders for 22 h/day. The findings indicated that sleep efficiency, slow wave sleep (SWS) and paradoxical sleep (PS) were decreased on the third and fourth days of unpredictable shocks compared to baseline while immobilization and swimming presented reduced sleep efficiency in all 4-day recordings. Swimming led to decreased SWS, whereas augmented PS was observed on the first day compared to baseline. Immobilization produced drastic alterations in sleep patterns since it reduced SWS during the 4 days and PS at days 1 to 4 in relation to baseline. Of all stressors, cold was the only one that did not result in any statistical differences in sleep pattern during the light periods. Regarding the effect of stress compared to baseline on the dark recordings, PS was higher during cold stress periods, whereas footshock increased PS on days 2 to 4 and swimming only on day 2. Immobilization decreased PS throughout the 4 days of the stress sessions. Thus, the data suggest that different stress modalities result in distinct sleep responses, with immobilization producing the most dramatic alterations.

Chronic psychosocial stress differentially affects apoptosis in hippocampal subregions and cortex of the adult tree shrew.

We studied the effect of chronic psychosocial stress on cell death and volume changes in the tree shrew hippocampus. In situ end labelling (ISEL) identified low frequent but convincing apoptosis in many hippocampal subregions. Also in entorhinal cortex, apoptosis was found, generally at higher frequencies. After 28 days of chronic stress, apoptosis was significantly reduced in the CA1 stratum radiatum, whereas an increase was observed in the hilus (P < 0.04). With all subregions taken together, the hippocampus showed a decrease, whereas in the cortex, an increase in apoptosis was found after stress (P < 0.04). In a parallel and similar chronic stress study, post mortem morphometry of the same brain regions was performed, revealing mild decreases (7.6%) in entire hippocampal volume. We conclude that (i) low frequent apoptosis occurs throughout the adult tree shrew brain, and (ii) 28 days of chronic stress differentially affects its occurrence in distinct hippocampal subregions and entorhinal cortex. As previous stereological investigations failed to detect any loss in the principal neuronal layers, psychosocial stress, therefore, must affect other (structural) parameters like dendritic tree, interneurons, neurogenesis, or glia.

Effects of chronic stress on anterior pituitary and brain corticotropin-releasing factor receptors

[ 125I]Corticotropin-releasing factor binding was measured in membrane homogenates prepared from the anterior pituitary, frontal cortex, motor cortex, somatosensory cortex, mesolimbic area (olfactory tubercle and nucleus accumbens), caudate putamen, hypothalamus, midbrain, and cerebellum from control and chronically stressed rats. The stressor consisted of 3 or 14 days of around-the-clock intermittent foot-shock (approximately one trial per 5-min frequency) that could be avoided or escaped on 90% of the trials presented by pulling a ceiling chain. Plasma corticosterone levels were almost doubled in stressed rats following 3 days of chronic stress and remained significantly elevated in rats stressed for 14 days as compared to controls. Plasma corticotropin levels were similar in controls and stressed animals in both the 3- and 14-day experiments. [ 125I]Corticotropin-releasing factor binding was decreased in anterior pituitary and frontal cortex following 3 ways of chronic stress; binding affinity of anterior pituitary membranes was not different between control and stressed animals. [ 125I]Corticotropin-releasing factor binding was similar in control and 3-day-stressed animals in the other brain areas examined. After 14 days of chronic stress, hypothalamic [ 125I]corticotropin-releasing factor binding was decreased in stressed rats as compared to control animals but no other differences were seen. The decrease in the apparent number of anterior pituitary corticotropin-releasing factor receptors following 3 days of stress may be due, in part, to increased plasma corticosterone levels and/or increased corticotropin-releasing factor secretion during that time. The downregulation of frontal cortex and hypothalamic corticotropin-releasing factor receptors, however, is more likely to be primarily in response to sustained stress-induced corticotropin-releasing factor release at those sites.

Adrenal responsiveness in domestic sheep (Ovis aries) to acute and chronic stressors as predicted by remote monitoring of cardiac frequency

The concept of stress and the general adaptive syndrome as advanced by Hans Selye has received considerable attention during the past decade primarily in its interpretation of physiological changes associated with chronic stress. Our work with domestic sheep (Ovis aries) habituated to stalls and fitted with halters carrying indwelling electrocardiogram leads and jugular vein cannulas allowed us to remotely test heart rate and blood cortisol responses of these animals to graded stressors. A radioimmunoassay was validated on domestic sheep plasma. We were unable to identify significant alterations of the adrenal response test by sheep exposed to synthetic adrenocorticotropic hormone after 34 days of chronic stress, suggesting neither adrenal exhaustion nor hypersensitivity. As an indicator of acute stress, we obtained a correlation coefficient of 0.91 between heart rate and blood cortisol, which suggests that heart rate has a strong potential of being a reliable predictor of cortisol values. With a regression equation, the heart rate of observed free-living sheep monitored by telemetry could be used to predict plasma cortisol levels and that, in turn, to predict potential stress-induced changes in animal production, including immunity.

Rapid increase of phenylethanolamine N-methyltransferase by environmental stress in an inbred mouse strains.

The activity of-phenylethanolamine N-methyltransferase in mice of the C57Bl/Ka strain was determined after a 4 degrees C stress. The enzyme activity increased 1.2-fold at the end of 3 hours and by 1.4-fold by the end of 6 hours of the stress. The results are in contrast to those from other species with intact animals in which the enzyme changes only after several days of chronic stress. Cycloheximide prevents the rise in enzyme activity, suggesting the increase may be due to protein synthesis. The increase may provide a model system for studying regulation of catecholamine biosynthetic enzymes.