Post-stress glucose consumption facilitates hormesis and resilience to severe stress.

Oral ingestion of a glucose solution following severe stress is a simple and effective way of preventing several of the negative sequelae of stress in rats. Similar resilience is obtained through hormetic training - pre-exposure to mild-to-moderate stress prior to severe stress. Here, we examined whether hormetic training is facilitated when a glucose solution is available following each hormetic training session. In Experiment 1, all rats were pre-exposed to a 30 min hormetic session of 25 inescapable tailshocks on each of 3 days. The schedule or hormesis differed between groups. The hormetic sessions occurred on either 3 consecutive days or with an interpolated day of rest between each hormetic session. Furthermore, in each of these conditions, one group had access to water and one group had access to a 40% glucose solution immediately after each hormetic session to complete a 2x2 factorial design. All groups were exposed to 100 inescapable tailshocks on the day following the end of hormetic training. Shuttle-escape testing occurred 24 h later. In Experiment 2, rats received two consecutive days of 100 inescapable tailshocks. Water or glucose was available following each session. Testing occurred 24 h after the second shock exposure. Experiment 1 replicated previous findings that rats exposed to hormetic training with interpolated rest did not show exaggerated fear responding or shuttle-escape deficits that normally result from 100 inescapable tailshocks, but training was ineffective if no rest was given between stress sessions. However, all post-stress glucose groups showed an elimination of helpless behavior. In Experiment 2, it was revealed that even 100 tailshocks can be made hormetic by post-stress glucose consumption.

Rigid patterns of effortful choice behavior after acute stress in rats.

Physical effort is a common cost of acquiring rewards, and decreased effort is a feature of many neuropsychiatric disorders. Stress affects performance on several tests of cognition and decision making in both humans and nonhumans. Only a few recent reports show impairing effects of stress in operant tasks involving effort and cognitive flexibility. Brain regions affected by stress, such as the medial prefrontal cortex and amygdala, are also implicated in mediating effortful choices. Here, we assessed effort-based decision making after an acute stress procedure known to induce persistent impairment in shuttle escape and elevated plasma corticosterone. In these animals, we also probed levels of polysialyted neural cell adhesion molecule (PSA-NCAM), a marker of structural plasticity, in medial frontal cortex and amygdala. We found that animals that consistently worked for high magnitude rewards continued to do so, even after acute shock stress. We also found that PSA-NCAM was increased in both regions after effortful choice experience but not after shock stress alone. These findings are discussed with reference to the existing broad literature on cognitive effects of stress and in the context of how acute stress may bias effortful decisions to a rigid pattern of responding.

Parameters of hormetic stress and resilience to trauma in rats.

Hormesis is the process by which small stresses build resilience to large stresses. We pre-exposed rats to various parameters of mild-to-moderate stress prior to traumatic stress in the present experiments to assess the potential benefits of hormetic training on resilience to traumatic, uncontrollable stress. Rats underwent varying stress pre-training parameters prior to exposure to uncontrollable traumatic stress in the learned helplessness procedure. The ability to prevent the exaggerated fear responding and escape deficits that normally follow experience with traumatic stress were used as a measure of the benefits of hormetic training. Four experiments examined the effects of number of training sessions, stressor severity and pattern of rest between pre-training stress sessions. Repeated exposure to mild restraint stress or moderate shock stress eliminated both the enhanced fear conditioning and shuttle-escape deficits that result from exposure to traumatic, inescapable shock. The pattern of rest did not contribute to resilience when the pre-exposure stressor was mild, but was vital when the pre-exposure stressor was moderate, with an alternation of stress and rest being the most effective procedure. The data also suggest that the level of resilience may increase with the number of pre-exposure sessions.

Post-Stress Fructose and Glucose Ingestion Exhibit Dissociable Behavioral and Physiological Effects.

An acute traumatic event can lead to lifelong changes in stress susceptibility and result in psychiatric disease such as Post-Traumatic Stress Disorder (PTSD). We have previously shown that access to a concentrated glucose solution for 24 hours beginning immediately after trauma decreased stress-related pathology in the learned helplessness model of PTSD and comorbid major depression. The current study sought to investigate the peripheral physiological effects of post-stress glucose consumption. We exposed 128 male Sprague-Dawley rats to inescapable and unpredictable 1-milliamp electric tail shocks or simple restraint in the learned helplessness procedure. Rats in each stress condition had access to a 40% glucose solution, 40% fructose solution, or water. Blood and liver tissue were extracted and processed for assay. We assessed corticosterone, corticosteroid-binding globulin (CBG), glucose, and liver glycogen concentrations at various time points following stress. We found that rats given access to glucose following exposure to traumatic shock showed a transient rise in blood glucose and an increase in liver glycogen repletion compared to those that received water or fructose following exposure to electric shock. We also found that animals given glucose following shock exhibited reduced free corticosterone and increased CBG compared to their water-drinking counterparts. However, this difference was not apparent when glucose was compared to fructose. These data suggest that post-stress glucose prophylaxis is likely not working via modulation of the HPA axis, but rather may provide its benefit by mitigating the metabolic challenges of trauma exposure.

Enhancing brain adenosine signaling with the nucleoside transport blocker NBTI (S-(4-nitrobenzyl)-6-theoinosine) mimics the effects of inescapable shock on later shuttle-escape performance in rats.

Experience with unsignaled, inescapable shock represents a profound challenge to brain metabolic function and physiology. The authors have argued that behavioral impairment following this traumatic stress is a consequence of enhanced brain adenosine signaling, which promotes metabolic recovery by profoundly inhibiting neural activation. The authors tested this hypothesis by artificially increasing extracellular brain adenosine concentration by blocking uptake transport with NBTI in rats given only restraint stress in five experiments. NBTI impaired shuttle-escape performance in the manner of inescapable shock in a dose-dependent manner and acted synergistically with an ineffective number of inescapable shocks to maximally impair test performance. These deficits produced by inescapable shock and NBTI were reversed by the nonselective adenosine receptor antagonist caffeine, and the highly selective A-sub(2A) receptor antagonist CSC (8-(3-chloro-styrl)caffeine). The highly selective A-sub-1 receptor antagonist DPCPX (8-Cyclopentyl-1,3-Dipropylxanthine) failed to improve performance in rats preexposed to inescapable shock or pretreated with NBTI. These data suggest that enhanced adenosine signaling at a brain A-sub(2A) receptor impairs escape performance following inescapable shock in the learned helplessness paradigm.

Brief flight to a familiar enclosure in response to a conditional stimulus in rats.

The authors observed brief, directed movement to a familiar enclosure in rats to determine whether this behavior is part of a rat's defensive repertoire when exposed to a conditional-fear stimulus. In Experiment 1, upon exposure to the compound conditional-fear stimulus of tone and light, only rats that received paired presentations of the conditional stimuli and shock fled into a small, familiar enclosure where they then froze. Rats that had received unpaired presentations did not enter the enclosure in significant amounts when later tested. In Experiment 2, the authors observed rats' freezing and use of either a familiar or an unfamiliar enclosure when tested with a conditional-fear stimulus. Rats tested with a familiar enclosure entered it more quickly than did rats without prior exposure to the enclosure. Freezing was greatest when both training and testing environments were similar with respect to access to the enclosure. The results of these 2 experiments support the idea that brief, directed flight in rats is a component of the postencounter stage of predatory imminence (M. S. Fanselow & L. S. Lester, 1988) and is compatible with freezing.

Adenosine signaling in reserpine-induced depression in rats.

A single, 6 mg/kg intraperitoneal injection of reserpine increased floating time during forced swim testing 24h after administration in rats in five experiments. Although such behavioral depression traditionally is attributed to drug-induced depletion of brain monoamines, we examined the potential contribution of adenosine signaling, which is plausibly activated by reserpine treatment and contributes to behavioral depression in other paradigms. Whereas peripheral administration of the highly selective A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (0.5, 1.0, or 5.0mg/kg i.p.) 15 min before swim testing failed to improve performance in reserpine-treated rats, swim deficits were completely reversed by 7 mg/kg of the nonselective receptor antagonist caffeine. Performance deficits were also reversed by the nonselective A2 antagonist 3,7-dimethylxanthine (0, 0.5, 1.0mg/kg i.p.), and the highly selective A2A receptor antagonist (CSC: 8-(3 chlorostyral)caffeine) (0.01, 0.1, or 1.0mg/kg i.p.) in a dose-dependent manner. The highly selective A2B antagonist alloxazine had no beneficial effect on swim performance at any dose under study (0.1, 1.0, and 5.0mg/kg i.p.).

Avoidance expression in rats as a function of signal-shock interval: strain and sex differences.

Inbred Wistar Kyoto (WKY) rats express inhibited temperament, increased sensitivity to stress, and exaggerated expressions of avoidance. A long-standing observation for lever press escape/avoidance learning in rats is the duration of the warning signal (WS) determines whether avoidance is expressed over escape. Outbred female Sprague-Dawley (SD) rats trained with a 10-s WS efficiently escaped, but failed to exhibit avoidance; avoidance was exhibited to a high degree with WSs longer than 20-s. We examined this longstanding WS duration function and extended it to male SD and male and female WKY rats. A cross-over design with two WS durations (10 or 60 s) was employed. Rats were trained (20 trials/session) in four phases: acquisition (10 sessions), extinction (10 sessions), re-acquisition (8 sessions) and re-extinction (8 sessions). Consistent with the literature, female and male SD rats failed to express avoidance to an appreciable degree with a 10-s WS. When these rats were switched to a 60-s WS, performance levels in the initial session of training resembled the peak performance of rats trained with a 60-s WS. Therefore, the avoidance relationship was acquired, but not expressed at 10-s WS. Further, poor avoidance at 10-s does not adversely affect expression at 60-s. Failure to express avoidance with a 10-s WS likely reflects contrasting reinforcement value of avoidance, not a reduction in the amount of time available to respond or competing responses. In contrast, WKY rats exhibited robust avoidance with a 10-s WS, which was most apparent in female WKY rats. Exaggerated expression of avoidances by WKY rats, especially female rats, further confirms this inbred strain as a model of anxiety vulnerability.

Helplessness and escape performance: glutamate-adenosine interactions in the frontal cortex.

Adenosine has been implicated as a proximate mediator of escape deficits in the learned helplessness paradigm, suggesting that neuronal overactivation-a typical precursor to adenosine release-precedes the inescapable shock-induced impairment (T. R. Minor, W. C. Chang, & J. L. Winslow, 1994). In the present experiments, glutamate (100 microg) injection into the rat frontal cortex produced a deficit in escape performance. Pretest treatment with the adenosine receptor antagonist caffeine (7 mg/kg ip) reversed the effect of glutamate when infused 1 hr. but not 72 hr, after glutamate injection. Finally, microinjection of 2-amino-5-phosphonovaleric acid (5 ng) into the frontal cortex prior to inescapable shock prevented the escape deficit. These findings are consistent with the involvement of N-methyl-D-aspartate receptor activation in the frontal cortex in the helplessness effect.

The L-type calcium channel blocker nimodipine mitigates "learned helplessness" in rats.

We assessed the effect of nimodipine, an L-type calcium channel blocker, on the escape deficit induced by prior exposure to inescapable shock in rats in four experiments. In Experiment 1, we injected rats at each of three time points (i.e., before shock exposure, after shock exposure, and before shuttle escape testing) with one of four doses of nimodipine (0, 0.5, 2.5, 5.0 mg/kg). The 5.0-mg/kg dose was most effective, acting to reduce shuttle escape latencies of inescapably shocked rats to a level comparable with nonshocked controls. No benefit occurred in Experiment 2, however, when nimodipine was administered at only one of the three time points used in the first experiment. Moreover, escape performance did not improve when rats received injections of nimodipine on the 2 days prior the experiment, and then one additional injection at one of the three time points identified above in Experiment 3. Finally, administration of nimodipine at two of the three time points did improve escape responding, but only when injected immediately prior both to shock exposure and the shuttle escape test.

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.

Cytokine-purine interactions in behavioral depression in rats.

This paper reviews recent findings from our laboratories concerning metabolic and immune mediators of behavioral depression in rats. Specifically, a single injection of 6 mg/kg of reserpine substantially increases behavioral depression, as evidenced by an increase in the amount of time spent floating by independent groups of rats tested for swim performance at various times during the next week. The behavioral impairment consists of two components. An early component emerges one hour after reserpine treatment and persists for about 24 hours. The deficit is not reversed by intracranial ventricular infusion of the receptor antagonist for interleukin-1beta (IL-1beta). A second, late-component deficit appears approximately 48 hours after reserpine treatment and recovers within a week. Late-component depression is reversed by central infusion of the IL-1beta receptor antagonist, and is mimicked by central infusion of the proinflammatory cytokine. Importantly, both early and late components of reserpine-induced depression and IL-1beta induced depression are reversed by a systemic injection of the highly selective A2A adenosine receptor antagonist 8-(3-Chlorostyryl) caffeine. These data are discussed in terms of the overlap in the conservation-withdrawal reaction during sickness, traumatic stress, and major depression and the regional contribution of purines and cytokines to the organization of this reaction in the brain.

The effect of glucose on hippocampal-dependent contextual fear conditioning.

BACKGROUND: The metabolic challenge of trauma disrupts hippocampal functioning, which is necessary for processing the complex co-occurring elements comprising the traumatic context. Poor contextual memory of trauma may subsequently contribute to intrusive memories and overgeneralization of fear. Glucose consumption following trauma may be a means to protect hippocampal functioning and contextual fear learning. This study experimentally examined the effect of glucose on hippocampal-dependent contextual learning versus cued fear learning in humans. METHODS: Forty-two male participants underwent cued conditioning with an unconditional stimulus (US) (shock) paired with a discrete conditional stimulus (geometric shape) and context conditioning (requiring hippocampal processing) with a US unpredictably paired with a background context (picture of room). Participants were then blindly randomized to consume either a 25 g glucose or sweet-tasting placebo drink and returned for a test phase 24 hours later. Measures included acoustic startle response, US expectancy, blood glucose levels, and arousal ratings. RESULTS: The glucose group showed superior retention of hippocampal-dependent contextual learning at test relative to the placebo group, as demonstrated by acoustic startle response and US expectancy ratings. Glucose and placebo groups did not differ on any measure of cued fear learning at test. CONCLUSIONS: This study provides experimental evidence that in mildly stressed humans postconditioning glucose consumption improves retention of hippocampal-dependent contextual learning but not cued learning. Ultimately, glucose consumption following trauma may be a means of improving learning about the traumatic context, thereby preventing subsequent development of symptoms of posttraumatic stress.

Modifying resilience mechanisms in at-risk individuals: a controlled study of mindfulness training in Marines preparing for deployment.

OBJECTIVE: Military deployment can have profound effects on physical and mental health. Few studies have examined whether interventions prior to deployment can improve mechanisms underlying resilience. Mindfulness-based techniques have been shown to aid recovery from stress and may affect brain-behavior relationships prior to deployment. The authors examined the effect of mindfulness training on resilience mechanisms in active-duty Marines preparing for deployment. METHOD: Eight Marine infantry platoons (N=281) were randomly selected. Four platoons were assigned to receive mindfulness training (N=147) and four were assigned to a training-as-usual control condition (N=134). Platoons were assessed at baseline, 8 weeks after baseline, and during and after a stressful combat training session approximately 9 weeks after baseline. The mindfulness training condition was delivered in the form of 8 weeks of Mindfulness-Based Mind Fitness Training (MMFT), a program comprising 20 hours of classroom instruction plus daily homework exercises. MMFT emphasizes interoceptive awareness, attentional control, and tolerance of present-moment experiences. The main outcome measures were heart rate, breathing rate, plasma neuropeptide Y concentration, score on the Response to Stressful Experiences Scale, and brain activation as measured by functional MRI. RESULTS: Marines who received MMFT showed greater reactivity (heart rate [d=0.43]) and enhanced recovery (heart rate [d=0.67], breathing rate [d=0.93]) after stressful training; lower plasma neuropeptide Y concentration after stressful training (d=0.38); and attenuated blood-oxygen-level-dependent signal in the right insula and anterior cingulate. CONCLUSIONS: The results show that mechanisms related to stress recovery can be modified in healthy individuals prior to stress exposure, with important implications for evidence-based mental health research and treatment.

Vulnerability factors in anxiety: Strain and sex differences in the use of signals associated with non-threat during the acquisition and extinction of active-avoidance behavior.

Rats that exhibit a behaviorally inhibited temperament acquire active-avoidance behaviors quicker, and extinguish them slower, than normal outbred rats. Here we explored the contribution of stimuli that signal periods of non-threat (i.e. safety signals) in the process of acquiring active-avoidance behavior. Utilizing a discrete lever-press escape-avoidance protocol, outbred Sprague-Dawley (SD) rats and inbred, behaviorally inhibited, Wistar-Kyoto (WKY) rats were tested under conditions where a flashing light was either presented or not during periods of non-threat (the inter-trial interval, ITI). For males, we found the absence of the ITI-signal slowed the acquisition of avoidance behavior selectively in WKY rats. However, extinction of the avoidance behavior was not influenced by training with or without the ITI-signal; WKY males extinguished slower than SD males. For females, the presence of the ITI-signal did not affect acquisition in either strain. However, after training with the ITI-signal, females of both strains extinguished quicker in its absence than in its presence. In order to determine if facilitated acquisition of avoidance learning in male WKY rats was due to a paradigm-independent influence of the visual stimulus used as ITI-signal upon associative learning, we conducted eyeblink conditioning in the presence or absence of a similar visual stimulus. No differences in acquisition, as a function of this visual stimulus, were observed within the male WKY rats, but, as was observed in avoidance learning, male WKY rats extinguished slower than male SD rats. Thus, avoidance susceptibility for male WKY rats may be tied both to the presence of non-threat signals as well as a resistance to extinguish Pavlovian-conditioned associations. Female susceptibility to resist extinguishing avoidant behavior is discussed with respect to the possible role of stimuli serving as occasion setters for threat contexts.

Avoidance perseveration during extinction training in Wistar-Kyoto rats: an interaction of innate vulnerability and stressor intensity.

Given that avoidance is a core feature of anxiety disorders, Wistar-Kyoto (WKY) rats may be a good model of anxiety vulnerability for their hypersensitivity to stress and trait behavioral inhibition. Here, we examined the influence of strain and shock intensity on avoidance acquisition and extinction. Accordingly, we trained WKY and Sprague-Dawley (SD) rats in lever-press avoidance using either 1.0-mA or 2.0-mA foot-shock. After extinction, neuronal activation was visualized by c-Fos for overall activity and parvalbumin immunoreactivity for gamma-aminobutyric acid (GABA) neuron in brain areas linked to anxiety (medial prefrontal cortex and amygdala). Consistent with earlier work, WKY rats acquired lever-press avoidance faster and to a greater extent than SD rats. However, the intensity of foot shock did not differentially affect acquisition. Although there were no differences during extinction in SD rats, avoidance responses of WKY rats trained with the higher foot shock perseverated during extinction compared to those WKY rats trained with lower foot shock intensity or SD rats. WKY rats trained with 2.0-mA shock exhibited less GABAergic activation in the basolateral amygdala after extinction. These findings suggest that inhibitory modulation in amygdala is important to ensure successful extinction learning. Deficits in avoidance extinction secondary to lower GABAergic activation in baslolateral amygdala may contribute to anxiety vulnerability in this animal model of inhibited temperament.

Biochemical and anatomical substrates of depression and sickness behavior.

This paper reviews recent research on the contribution of the proinflammatory cytokine interleukin-1 (IL- 1) and the purine nucleoside adenosine in mediating behavioral depression and related symptoms of conservation-withdrawal in animal models of both major depression and illness. Activation of brain IL- 1 receptors appears to contribute to conservation withdrawal symptoms in animals treated with reserpine or lipopolysaccharide, suggesting a common underlying mechanism. Moreover, brain cytokine signaling is capable of recruiting adenosine signaling at adenosine A2A receptors, which directly mediate symptoms of behavioral depression. The adenosine receptors densely populate spiny GABAergic neurons in the striopallidal tract in the striatum and form part of an A2A/D2/mGLU receptor complex. Activation of these A2A receptors functionally uncouples dopamines excitatory motivation influence from ongoing behavior, leading to a state of conservation-withdrawal, and antagonism of the ventral medial striatum A2A receptors in reserpinated rats relieves symptoms of behavioral depression.

Cytokine-purine interactions in traumatic stress, behavioral depression, and sickness.

This paper reviews recent research on the contribution of the proinflammatory cytokine interleukin-1beta (IL-1beta) and the purine nucleoside adenosine in mediating behavioral depression and related symptoms of conservation-withdrawal in animal models of traumatic stress, major depression, and illness. Activation of brain IL-1beta receptors appears to contribute to conservation-withdrawal by exacerbating the immediate impact of a stressor or by enhancing and prolonging the overall reaction. Moreover, brain cytokine signaling is capable of recruiting adenosine signaling at A(2A), which directly mediates symptoms of behavioral depression. The adenosine receptors densely populate spiny GABAergic neurons in the striopallidal tract in the striatum and form part of an A(2A)/D(2)/mGLU receptor complex. Activation of these A(2A) receptors functionally uncouples dopamine's excitatory motivation influence on ongoing behavior, leading to a state of conservation-withdrawal.

Brain IL-1 beta was involved in reserpine-induced behavioral depression in rats.

AIM: To investigate the mechanism of brain interleukin-1 beta (IL-1 beta) in reserpine-induced behavioral depression in rats. METHODS: Porsult swim test was used in the measurement of depressive behavior and ELISA was used in measurement of brain IL-1 beta. RESULTS: Intraperitoneal injection of reserpine (0, 4, 6, and 8 mg/kg, ip) increased floating time in the Porsult swim test in a dose-and time-dependent manner in rats. Intracerebroventricular injection (icv) of IL-1 beta receptor antagonist (IL-1ra, 6 mg/kg) blocked the increment of floating time in Porsult swim test at 48 and 72 h after reserpine injection, but not at 1 and 24 h after injection. Brain IL-1 beta increased after reserpine treatment in posterior cortex, hippocampus, and hypothalamus. The increase of IL-1 beta concentration starts at 24 hours after injection of reserpine and reached the peak at 48 h. CONCLUSION: Reserpine induced behavioral depression partially via brain interleukin-1 beta generation.