Timing is everything: Anticipatory stress dynamics among cortisol and blood pressure reactivity and recovery in healthy adults

Psychological states of anticipation modulate biological stress responsivity. While researchers generally investigate how subjective distress corresponds to the magnitude of stress reactivity, physiological recovery after acute stressors must also be considered when investigating disease vulnerabilities. This study assessed whether anticipatory stress would correspond to stress reactivity and recovery of salivary cortisol and blood pressure levels in response to a well-validated psychosocial stressor. Thirty participants (63% female; mean?±?SEM age 45.4?±?2.12 years) were exposed to the Trier Social Stress Test (TSST) consisting of a public speech and mental arithmetic. Ten salivary cortisol samples and systolic and diastolic blood pressure recordings were collected at time points spanning 50?min before and up to 50?min after stress exposure. These data were transformed into parameters representing stress reactivity (area under the curve) and stress recovery (percent change). The Primary Appraisal Secondary Appraisal scale assessed anticipatory stress before exposure to the TSST. Our results revealed that increased anticipatory stress predicted increased stress reactivity for cortisol (p?=?0.009) but not blood pressure. For stress recovery, increased anticipatory stress predicted greater decrements of cortisol concentration (p?=?0.015) and blood pressure (p?=?0.039), even when controlling for total systemic "output" by incorporating baseline activity. This efficient shutdown of stress responses would have otherwise been ignored by solely investigating reactive increases. These findings underscore the importance of measuring multiple dynamic parameters such as recovery when investigating physiological stress response patterns as a function of psychosocial factors.     

Neuronal nitric oxide synthase gene inactivation reduces the expression of vasopressin in the hypothalamic paraventricular nucleus and of catecholamine biosynthetic enzymes in the adrenal gland of the mouse

The impact of a lifelong absence of the neuronal nitric oxide synthase (nNOS) in the neuroendocrine stress response was investigated in nNOS knockout (KO) and wild type (WT) mice under basal conditions and in response to forced swimming. In the hypothalamic paraventricular nucleus oxytocin and corticotropin-releasing-hormone mRNA levels did not differ between these genotypes under resting conditions, whereas vasopressin mRNA levels were significantly lower in nNOS KO than in WT animals. Also, in the adrenal glands basal levels of tyrosine hydroxylase protein, the rate-limiting enzyme for catecholamine biosynthesis, and of phenylethanolamine N-methyltransferase, which converts norepinephrine to epinephrine, were significantly reduced in nNOS KO mice. Plasma adrenocorticotropin, corticosterone, norepinephrine and epinephrine levels were similar in the KO and WT genotypes under resting conditions. In response to forced swimming, a similar increase in plasma adrenocorticotropin and corticosterone was observed in KO and WT animals. Stressor exposure triggered also an increased epinephrine release in WT animals, but did not significantly alter plasma epinephrine levels in KO mice. These data suggest that the chronic absence of nNOS reduces the capacity of epinephrine synthesising enzymes in the adrenal gland to respond to acute stressor exposure with an adequate epinephrine release.     

Gender Differences in Biobehavioral Aftereffects of Stress on Eating,Frustration, and Cardiovascular Responses1

Eating, persistence, and cardiovascular responses were evaluated after exposure to a 25‐min noise stressor with or without perceived control. Participants were healthy men (n = 29) and women (n = 34), aged 21 to 45 years. There were no group differences in cognitive task performance or blood pressure during the stressor. However, perceived control resulted in lower mean blood pressure and heart rate after cessation of the stressor for men and women. Women without perceived control displayed greater frustration levels following the stressor, and frustrated women ate more bland food than did nonfrustrated women. Perceived control and frustration did not affect food consumption among men following the stressor. These findings indicate that there are health‐relevant gender differences in biobehavioral responses that occur in the aftermath of stressor exposure. In addition, perceived control was especially important for women to attenuate the behavioral and biological effects of stressor exposure.     

The effects of metabolic stress and vagotomy on emotional learning in an animal model of anxiety

The aim of the present study is to evaluate the role of the blood glucose (BG) level in emotional learning in the elevated plus maze (EPM), an animal model of anxiety. In Experiment 1, male Wistar rats were submitted to different EPM trial lengths (1- or 5-min). Blood samples were withdrawn before and after the maze exploration, through a polyethylene cannula chronically implanted into the jugular vein. In Experiment 2, the animals received either saline or 2-deoxy-D-glucose, a glucoprivic drug (2-DG, 250 or 500 mg kg(-1)) by i.p. route, 30 min before a 5-min EPM exposure and were retested in the maze (Trial1/Trial2 EPM procedure) 24 h later. In an independent group of rats, blood samples were withdrawn 0, 5, 15, and 30 min after 2-DG administration, through the jugular vein, to determine BG. In Experiment 3, the animals underwent a vagotomy and were tested in a Trial1/Trial2 EPM procedure four weeks later. The results showed that rats exploring the EPM for 5 min displayed increased fear and higher hyperglycemia than those exploring the EPM for 1 min. In addition, rats submitted to 5-min EPM Trial1 length displayed higher level of fear on Trial2, as well as higher percentage of shortening of the %Open arm entries and %Open arm time from Trial1 to Trial2, which characterizes the occurrence of emotional learning. In contrast, rats previously vagotomized or treated with 2-DG (500 mg kg(-1)) showed the same level of fear on both EPM trials and a low percentage of shortening, from Trial1 to Trial2, of the %Open arm entries and %Open arm time, indicating poor emotional learning. The data is discussed regarding the role of glycaemia in emotional learning in the EPM.     

Influence of Regularity of Exposure to Chronic Stress on the Pattern of Habituation of Pituitary-Adrenal Hormones, Prolactin and Glucose

The effect of regularity of exposure to two different chronic stressors (noise or immobilization (IMO)) on the pattern of habituation of pituitary-adrenal (PA) hormones, prolactin and glucose was evaluated in adult male rats. Animals were chronically subjected to either regular or irregular time schedule of noise (30 min/day) or IMO (2 h/day) for two weeks. The day after the last stress session the rats were killed without stress or after having been subjected to 30 min of the homotypic stressor. Whereas regular noise did not affect food intake, body weight gain or adrenal weight, irregular noise decreased body weight gain and induced a moderate adrenal hypertrophy. In addition, previous daily exposure to regular but not to irregular noise reduced both prolactin and corticosterone responses to acute noise. In contrast, glucose response to acute noise was reduced after both regular and irregular exposure to chronic noise. Either regular or irregular exposure to chronic IMO decreased food intake and body weight and increased adrenal weight to the same extent. Likewise, no influence of regularity of exposure to chronic IMO on corticosterone and prolactin responses to acute IMO was observed. However, habituation of the ACTH response to acute IMO was observed in rats subjected to chronic regular IMO, but not in rats subjected to chronic irregular IMO. Finally, acute IMO-induced hyperglycemia diminished to the same extent after regular and irregular IMO. From these results we can conclude that: first, the process of habituation of the PA axis to chronic stress is greatly dependent upon factors such as regularity of exposure to the stressor and stressor intensity, and second, the influence of regularity on the pattern of habituation to a repeated stressor is dependent on the physiological variable we are dealing with.     

Serial stressors: prior exposure to a stressor modulates its later effectiveness on gastric ulceration and corticosterone release

Several studies have focused on the proactive effects of prior exposure to a stressor on ulcerogenic and endocrine responses to exposure to a similar final stressor. In the present study, we explored the effects of prior exposure of M?ll-Wistar rats to water-restraint stress on gastric ulceration and corticosterone responses to a final stressor. The number of prior exposures to water-restraint challenge was varied from one to four, and the durations of the prior exposures varied from 30 min to 2 h. The duration of the final stress challenge was 75 min. The two indices of the stress response to the final stressor challenge yielded different results. For neither index was the duration of prior exposures a significant factor in modulating the response to the final challenge. For gastric ulceration, those animals receiving one prior exposure, but not four, exhibited less gastric ulceration than animals receiving no prior exposure. These animals also developed less ulceration than those receiving four prior exposures. For corticosterone, those animals receiving four prior exposures exhibited a lower adrenocortical response to the final stressor than did animals receiving no prior exposure, or only one prior exposure. The results indicate the complexity of proactive effects of earlier stress experience on responses to a final stressor challenge.     

Acute and chronic stress effects on performance in a forced-swim task

The effects of uncontrollable stressors on performance in a subsequent forced-swim paradigm were assessed in mice. Uncontrollable shock initially induced behavioral invigoration; however, within 24 h of stressor application, swimming behavior was depressed relative to nonstressed mice. The controllability of the stressor did not influence the initial invigoration, being present among escapably shocked mice as well as among mice that received (yoked) inescapable shock. In contrast, the depression of responding evident 24 h after stressor application was related to the availability of behavioral coping methods. Finally, following repeated exposure to footshock there was no indication of adaptation to the behavioral changes ordinarily induced by acute shock stress. The data were related to the effects of uncontrollable stressors on escape performance, and with respect to the use of this preparation as an animal model of human depression.     

Acute stress persistently enhances estrogen levels in the female rat

Here we tested whether exposure to either tailshock or swim stress alters ovarian hormone levels, estrogen and progesterone, in females and whether the effects are persistent. Adrenal hormone levels were also measured in males and females. Estradiol levels were elevated in unstressed females during proestrus relative to females in other stages of estrous, and exposure to the stressors enhanced estradiol beyond basal levels. For females stressed during diestrus 2, estradiol levels were elevated immediately after stressor cessation and up to 24 hrs. Exposure to tailshock, but not swim-stress, transiently enhanced progesterone in females stressed during the stage of proestrus and estrus. Glucocorticoid levels were elevated in response to both stressors and were supraelevated in females under both basal and stress conditions relative to males, particularly in blood from females exposed to acute swim stress. These results indicate that exposure to a relatively acute stressful event immediately and persistently enhances serum estradiol and are discussed in the context of reports that exposure to the same stressors immediately and persistently impairs associative learning in the female rat.     

Chronic psychosocial stress induces visceral hyperalgesia in mice

Experimental and clinical evidence has shown that chronic stress plays an important role in the onset and/or exacerbation of symptoms of functional gastrointestinal disorders. Here, we aimed to investigate whether exposure to a chronic and temporally unpredictable psychosocial stressor alters visceral and somatic nociception as well as anxiety-related behaviour. In male C57BL/6J mice, chronic stress was induced by repeated exposure to social defeat (SD, 2?h) and overcrowding (OC, 24?h) during 19 consecutive days. Visceral and somatic nociception was evaluated by colorectal distension and a hot plate, respectively. The social interaction test was used to assess social anxiety. Mice exposed to psychosocial stress developed visceral hyperalgesia and somatic hypoalgesia 24?h following the last stress session. SD/OC mice also exhibited social anxiety-like behaviour. All these changes were also associated with physiological alterations, measured as a decreased faecal pellet output and hypothalamic-pituitary-adrenal (HPA) axis disruption. Taken together, these data confirm that this mouse model of chronic psychosocial stress may be useful for studies on the pathophysiological mechanisms underlying such stress-associated disorders and to further test potential therapies.     

Temporal relationships between plasma cortisol, corticosteroid-binding globulin (CBG), and the free cortisol index (FCI) in pigs in response to adrenal stimulation or suppression

The objective of this study was to document changes in plasma concentrations of total cortisol, porcine corticosteroid-binding globulin (pCBG), and the free cortisol index (FCI) in pigs over a 6-h period in response to adrenal stimulation or suppression. Twenty-four 8-week old pigs allotted in equal numbers were administered ACTH, dexamethasone or saline, and blood samples were collected every 15 min via an indwelling jugular catheter for 1 h prior to and 5 h following treatment. Total plasma cortisol increased in ACTH-treated pigs and decreased in dexamethasone-treated pigs within 0.25 and 0.5 h, respectively. In contrast, pCBG concentration was altered in an inverse fashion subsequent to the changes exhibited in total cortisol. FCI reflected the changes observed in total cortisol. These results further document the negative relationship that exists between circulating concentrations of plasma cortisol and pCBG, and illustrate that this association exists under conditions of acute stress in the pig.     

Negative affect as a predisposing factor for cortisol release after an acute stress--the impact of unpleasant priming

Glucocorticoids have a key role in stress responses. There are, however, substantial differences in cortisol reactivity among individuals. We investigated if affective trait and mood induction influence the reactivity to psychological stress in a group of 63 young adults, male (n=27) and female (n=36), aged ca. 21 years. On the experimental day the participants viewed either a block of pleasant or unpleasant pictures for 5 min to induce positive or negative mood, respectively. Then, they had 5 min to prepare a speech to be delivered in front of a video-camera. Saliva samples were collected to measure cortisol, and questionnaire-based affective scales were used to estimate emotional states and traits. Compared to basal levels, a cortisol response to the acute speech stressor was only seen for those who had first viewed unpleasant pictures and scored above the average on the negative affect scale. There were no sex differences. In conclusion, high negative affect associated with exposure to an unpleasant context increased sensitivity to an acute stressor, and was critical to stimulation of cortisol release by the speech stressor.     

Effects and aftereffects of stressor expectations

Is stressor exposure necessary to produce "stress" effects, or can these effects result from stressor anticipation alone? The present research explores whether it is possible to obtain "stress responses" during and after the period in which stressor exposure is being anticipated. In the first study, the expectation of submerging one's hand in ice water resulted in decreased frustration tolerance and increased blood pressure when compared with control groups not expecting this stressor. A second study replicated and extended these results to show that the expectation of control over the stressor ameliorates the negative impact of stressor expectation. The second study also examined the aftereffects of expectations. Particularly, it found that despite being relieved of the expectation that they would immerse their hand in ice water, subjects who had expected stressor exposure had decreased frustration tolerance when compared with either subjects who had expected a nonstressful procedure or those who had expected to have control over stressor termination. A third study, using noise as the expected stressor, replicated both the aftereffect of the anticipation period and the moderation of that effect by perceived control. The discussion (a) focuses on the implications of this work for understanding why aftereffects occur and (b) proposes that previously observed stressor exposure effects may in fact be postexpectation effects.     

Predictability of chronic intermittent stress: effects on sympathetic-adrenal medullary responses of laboratory rats

This experiment involved an examination of sympathetic-adrenal medullary responses of laboratory rats following exposure to chronic intermittent stress. Animals were assigned at random to one of three groups: (i) controls, handled briefly each day; (ii) restraint stress (RS), restrained for 30 min per day; or (iii) variable stress (VS), exposure to restraint, cold swim, or intermittent footshock during one of five time periods each day. On the 26th day, rats were prepared with chronic tail artery catheters for remote sampling of blood and direct measurement of mean arterial pressure and heart rate. On Day 28, rats of each group were exposed to 30 min of restraint stress and timed blood samples were collected and later analyzed for content of norepinephrine (NE) and epinephrine (EPI). VS rats gained significantly less body weight compared to control and RS rats. Basal measures of blood pressure and heart rate and of plasma NE and EPI were comparable for rats of the three groups. The plasma catecholamine responses to restraint stress on Day 28 were significantly reduced in RS and VS rats compared to first-time stressed controls. These findings suggest that predictability of the type of stressor and the time of its occurrence does not influence the pattern of diminished sympathetic-adrenal medullary responses of animals exposed to chronic intermittent stress.     

Aggression is suppressed by acute stress but induced by chronic stress: Immobilization effects on aggression, hormones, and cortical 5-HT1B/ striatal dopamine D2 receptor density

Although it has been established by a number of investigators that a variety of stressors are associated with the induction of aggressive behavior, two specific issues remain unanswered. First, it is unclear whether the contexts surrounding stressors (e.g., stressor length and chance of winning over opponents) change outcomes regarding aggressive behavior. Second, if a relationship exists between stress and aggressive behavior, altered levels of stress-related hormone (e.g., corticosterone [CORT]), as well as aggression-related biomarkers (e.g., testosterone [T], density of prefronto-cortical 5-HT(1B) receptor and striatal dopamine D(2) receptor [D2r]) may contribute to changes in aggressive behavior. Thus, we examined how immobilization (with a 1-, 5-, or 10-day exposure) would impact (1) a longitudinal course of aggression toward different-sized opponents, (2) levels of CORT and T, and (3) densities of 5-HT(1B) receptor (5-HT1Br) in the prefrontal cortex (PFC) and D2r in the striatum. It was found that, regardless of small or large opponents, a single 2-h exposure to immobilization reduced aggressive behavior (stress-suppressed aggression) over time, whereas repeated (10-day) exposure to immobilization escalated aggressive behavior (stress-induced aggression). These stress effects persisted up to 1 week of recovery from immobilization stress. Moreover, immobilized rats demonstrated elevated levels of T, but not CORT, as compared with controls. Finally, acute immobilization altered D2r densities in the shell of the nucleus accumbens, and chronic immobilization changed 5-HT1Br in the PFC, including the downregulation of 5-HT1Br densities in the right prelimbic and orbitolateral cortices. The potential relationships among stress, aggression, and 5-HT1Br/D2r roles are discussed.     

An Exploration of a Possible Physiological Explanation for Stressor Aftereffects1

The present study examined the relationship between performance deficits that occur following exposure to uncontrollable stressors and activation of the endogenous opioid system. Subjects were given an injection of saline or naloxone, an opioid antagonist, prior to exposure to 23 min of unpredictable intermittent bursts of noise or silence. Changes in mood were assessed following the injection and the stressor. Performance was measured during and after stressor exposure. Measures of performance following the stressor included a proofreading, an encoding, and a puzzle task. Results replicated previous work on the effects of noise on performance; exposure to noise was associated with performance deficits during and after the stressor. Although naloxone had no effect on performance following the noise, it was associated with greater performance deficits during exposure to the stressor. Further, although all subjects became more hostile, anxious, and depressed following exposure to noise or silence, naloxone improved mood before the stressor began.     

Stressor Controllability and Learned Helplessness Research in the United States: Sensitization and Fatigue Processes

Recent work in the learned helplessness paradigm suggests that neuronal sensitization and fatigue processes are critical to producing the behavioral impairment that follows prolonged exposure to an unsignaled inescapable stressor such as a series of electric tail shocks. Here we discuss how an interaction between serotonin (5-HT) and corticosterone (CORT) sensitizes GABA neurons early in the pretreatment session with inescapable shock. We propose that this process eventually depletes GABA, thus removing an important form of inhibition on excitatory glutamate transmission in the amygdala, hippocampus, and frontal cortex. When rats are re-exposed to shock during shuttle-escape testing 24 hrs later, the loss of inhibition (as well as other excitatory effects) results in unregulated excitation of glutamate neurons. This state of neuronal over-excitation rapidly compromises metabolic homeostasis. Metabolic fatigue results in compensatory inhibition by the nucleoside adenosine, which regulates neuronal excitation with respect to energy availability. The exceptionally potent form of inhibition associated with adenosine receptor activation yields important neuroprotective benefits under conditions of metabolic failure, but also precludes the processing of information in fatigued neurons. The substrates of adaptive behavior are removed; performance deficits ensue.     

Acute stress rapidly and persistently enhances memory formation in the male rat

Previous studies, as well as the present one, report that acute exposure to intermittent tailshocks enhances classical eyeblink conditioning in male rats when trained 24 h after stressor cessation. In Experiment 1, it was determined that the facilitating effect of stress on conditioning could also be obtained in response to a stressor of acute inescapable swim stress but not inescapable noise or the unconditioned stimulus of periorbital eyelid stimulation. These selective responses arose despite comparable enhancements of the stress-related hormone corticosterone in response to tailshocks, periorbital eyelid stimulation, noise stress, and supraelevation in response to swim stress. Although corticosterone is necessary for the enhanced learning in response to stress (Beylin & Shors, 1999), these results suggest that it is not sufficient. In addition, the results suggest that the enhancement is not dependent on common characteristics between the stressor and the conditioning stimuli (stimulus generalization). In Experiment 2, it was determined that the facilitating effect of the stressor on conditioning occurs within 30 min of stressor cessation. Thus, the mechanism responsible for facilitating memory formation is rapidly induced as well as persistently expressed. In Experiment 3, it was determined that exposure to the stressor does not enhance performance of the conditioned response after the response has been acquired. Thus, exposure to the stressor enhances the formation of new associations rather than affecting retention or performance of the motor response. These studies extend the circumstances under which stress is known to enhance associative learning and implicate neural mechanisms of memory enhancement that are rapidly induced and persistently expressed.     

Stressor controllability and learned helplessness research in the United States: Sensitization and fatigue processes

Recent work in the learned helplessness paradigm suggests that neuronal sensitization and fatigue processes are critical to producing the behavioral impairment that follows prolonged exposure to an unsignaled inescapable stressor such as a series of electric tail shocks. Here we discuss how an interaction between serotonin (5-HT) and corticosterone (CORT) sensitizes GABA neurons early in the pretreatment session with inescapable shock. We propose that this process eventually depletes GABA, thus removing an important form of inhibition on excitatory glutamate transmission in the amygdala, hippocampus, and frontal cortex. When rats are re-exposed to shock during shuttle-escape testing 24 hrs later, the loss of inhibition (as well as other excitatory effects) results in unregulated excitation of glutamate neurons. This state of neuronal over-excitation rapidly compromises metabolic homeostasis. Metabolic fatigue results in compensatory inhibition by the nucleoside adenosine, which regulates neuronal excitation with respect to energy availability. The exceptionally potent form of inhibition associated with adenosine receptor activation yields important neuroprotective benefits under conditions of metabolic failure, but also precludes the processing of information in fatigued neurons. The substrates of adaptive behavior are removed; performance deficits ensue.     

The effects of exposure to an acute naturalistic stressor on working memory, state anxiety and salivary cortisol concentrations

Exposure to an acute naturalistic stressor induces both psychological and physiological changes in humans. The two studies reported here explored the impact of exposure to an acute naturalistic stressor on state anxiety, working memory and HPA axis activation (salivary cortisol). In both experiments, ten healthy male participants were exposed to an acute naturalistic stressor, helicopter underwater evacuation training (HUET), and their physiological and behavioural responses before (first study) and after (second study) the stressor were compared to ten non-stressed controls. The results of both experiments showed that working memory performance was preserved during anticipation of an acute stressor, but impairments were observed immediately after stress exposure. Participants reported significantly higher state anxiety levels during anticipation and following stress exposure, whereas significant elevations in cortisol levels were only observed 25 min post exposure to stress, but not before or immediately after stress exposure. The results of both experiments demonstrated a dissociation between behavioural and biochemical measures and provided evidence for a dissociation of the effects of stress on cognitive and physiological measures depending on the time of testing, with cognitive impairments most evident following stress exposure.     

Accounting for Parcel-Allocation Variability in Practice: Combining Sources of Uncertainty and Choosing the Number of Allocations

Item parceling remains widely used under conditions that can lead to parcel-allocation variability in results. Hence, researchers may be interested in quantifying and accounting for parcel-allocation variability within sample. To do so in practice, three key issues need to be addressed. First, how can we combine sources of uncertainty arising from sampling variability and parcel-allocation variability when drawing inferences about parameters in structural equation models? Second, on what basis can we choose the number of repeated item-to-parcel allocations within sample? Third, how can we diagnose and report proportions of total variability per estimate arising due to parcel-allocation variability versus sampling variability? This article addresses these three methodological issues. Developments are illustrated using simulated and empirical examples, and software for implementing them is provided.