Footshock Stimuli Research Articles

Use Electroencephalogram Entropy as an Indicator to Detect Stress-Induced Sleep Alteration

An acute stressor can cause sleep disruptions. Electroencephalography (EEG) is one of the major tools to measure sleep. In rats, sleep stages are classified as rapid-eye movement (REM) sleep and non-rapid-eye movement (NREM) sleep, by different characteristics of EEGs. Sleep alterations after exposure to an acute stress are regularly determined by the power spectra of brain waves and the changes of vigilance stages, and they all depend on EEG analysis. Herein, we hypothesized that the Shannon entropy can be employed as an indicator to detect stress-induced sleep alterations, since we noticed that an acute stressor, the footshock stimulation, causes certain uniformity changes of the spectrograms during NREM and REM sleep in rats. The present study applied the Shannon entropy on three features of brain waves, including the amplitude, frequency, and oscillation phases, to measure the uniformities in the footshock-induced alterations of sleep EEGs. Our result suggests that the footshock stimuli resulted in a smoother and uniform amplitude as well as varied frequencies of EEG waveforms during REM sleep. In contrast, the EEGs during NREM sleep exhibited a smoother, but less uniform, amplitude after the footshock stimuli. The result depicts the change property of brain waves after exposure to an acute stressor and, also, demonstrates that the Shannon entropy could be used to detect EEG alteration in sleep disorders.

Hypocretin in median raphe nucleus modulates footshock stimuli-induced REM sleep alteration

Stress is one of major factors that cause sleep problems. Hypocretin represents a stress-related neuropeptide and is well known in maintaining physiological wakefulness. The hypocretinergic neurons originate in the lateral hypothalamic area (LHA) and transmit to several brain regions, including the median raphe nuclei (MRNs). The MRNs modulate both fear responses and sleep-wake activity; however, it remains unclear whether stress alters the levels of hypocretin to regulate MRNs and consequently disrupt sleep. In this paper, we employed the inescapable footshock stimuli (IFS) as a stressor and hypothesized that the IFS-induced sleep disruption is mediated by increased hypocretins in the MRNs. Our results demonstrate that the concentrations of hypocretin in the hypothalamus increased after IFS. Rapid eye movement (REM) sleep was reduced after footshock, and microinjection of non-selective hypocretin receptor antagonist TCS-1102 into the MRNs blocked the IFS-induced decrease of REM sleep. Furthermore, administration of hypocretins into the MRNs mimicked the IFS-induced REM sleep reduction. These results conclude that the increased levels of hypocretins in the MRNs mediate the IFS-induced REM sleep reduction.

Early postnatal stress alters extracellular signal‐regulated kinase signaling in the corticolimbic system modulating emotional circuitry in adult rats

The present study elucidated whether early life stress alters the extracellular signal-regulated kinase (ERK) pathway that underlies fear retrieval and fear extinction based on a contextual fear conditioning paradigm, using a juvenile stress model. Levels of phospho-ERK (pERK), the active form of ERK, increased after fear retrieval in the hippocampal CA1 region but not in the medial prefrontal cortex (mPFC). ERK activation in the CA1 following fear retrieval was not observed in adult rats who received aversive footshock (FS) stimuli during the second postnatal period (2wFS), which exhibited low levels of freezing. In fear extinction, pERK levels in the CA1 were increased by repeated extinction trials, but they were not altered after extinction retrieval. In contrast, pERK levels in the mPFC did not change during extinction training, but were enhanced after extinction retrieval. These findings were compatible in part with electrophysiological data showing that synaptic transmission in the CA1 field and mPFC was enhanced during extinction training and extinction retrieval, respectively. ERK activation in the CA1 and mPFC associated with extinction processes did not occur in rats that received FS stimuli during the third postnatal period (3wFS), which exhibited sustained freezing behavior. The repressed ERK signaling and extinction deficit observed in the 3wFS group were ameliorated by treatment with the partial N-methyl-D-aspartate receptor agonist D-cycloserine. These findings suggest that early postnatal stress induced the downregulation of ERK signaling in distinct brain regions through region-specific regulation, which may lead to increased behavioral abnormalities or emotional vulnerabilities in adulthood.

P.1.c.031 Role of CB1 receptors in the basolateral amygdala in conditioned aversion, associated analgesia and neuronal activation

Agmatine, a noncompetitive N-methyl-d-aspartate (NMDA) antagonist, was examined for its role in water maze place learning, contextual and auditory-cued (discrete) fear learning and conditioned taste aversion learning, when administered systemically. Male Wistar rats were given saline or 1, 5, 10 or 50 mg/kg agmatine ip 20 min prior to or 30 min following daily training sessions in a hidden-platform (place learning) water maze task. Agmatine did not affect latencies to find the hidden platform or preference for the training quadrant during probe trials. When administered 20 min prior to contextual or auditory-cued fear-conditioning sessions, these doses of agmatine evoked a linear dose-dependent impairment in the magnitude of learned fear to the contextual stimuli when assessed during extinction trials 24 h later, but had no effect on the magnitude of learned fear to the auditory stimulus. Inferences of baseline motor activity and ability to respond to the presentation of footshock stimuli were not affected by the treatment. Injections of 50 mg/kg agmatine concurrently with a malaise-evoking agent following presentations to a novel sucrose solution abolished learned taste aversions; this agent did not evoke conditioned taste aversions alone. These studies indicate that systemically administered agmatine selectively impairs behavioral inferences of specific types of learning and memory.

Norepinephrine transporter-deficient mice respond to anxiety producing and fearful environments with bradycardia and hypotension

The study of anxiety and fear involves complex interrelationships between psychiatry and the autonomic nervous system. Altered noradrenergic signaling is linked to certain types of depression and anxiety disorders, and treatment often includes specific transporter blockade. The norepinephrine transporter is crucial in limiting catecholaminergic signaling. Norepinephrine transporter-deficient mice have increased circulating catecholamines and elevated heart rate and blood pressure. We hypothesized, therefore, that reduced norepinephrine clearance would heighten the autonomic cardiovascular response to anxiety and fear. In separate experiments, norepinephrine transporter-deficient (norepinephrine transporter −/−) mice underwent tactile startle and trace fear conditioning to measure hemodynamic responses. A dramatic tachycardia was observed in norepinephrine transporter −/− mice compared with controls following both airpuff or footshock stimuli, and pressure changes were also greater. Interestingly, in contrast to normally elevated home cage levels in norepinephrine transporter-deficient mice, prestimulus heart rate and blood pressure were actually higher in norepinephrine transporter +/+ animals throughout behavioral testing. Upon placement in the behavioral chamber, norepinephrine transporter-deficient mice demonstrated a notable bradycardia and depressor effect that was more pronounced in females. Power spectral analysis indicated an increase in low frequency oscillations of heart rate variability; in mice, suggesting increased parasympathetic tone. Finally, norepinephrine transporter −/− mice exhibited sexual dimorphism in freeze behavior, which was greatest in females. Therefore, while reduced catecholamine clearance amplifies immediate cardiovascular responses to anxiety- or fear-inducing stimuli in norepinephrine transporter −/− mice, norepinephrine transporter deficiency apparently prevents protracted hemodynamic escalation in a fearful environment. Conceivably, chronic norepinephrine transporter blockade with transporter-specific drugs might attenuate recognition of autonomic and somatic distress signals in individuals with anxiety disorders, possibly lessening their behavioral reactivity, and reducing the cardiovascular risk factors associated with persistent emotional arousal.

Anxiety-related behavior in juvenile stroke-prone spontaneously hypertensive rats — an animal model of attention-deficit/hyperactivity disorder

Attention-deficit/hyperactivity disorder (AD/HD) is defined as a developmental disorder, manifested by deficit sustained attention (inattention), and/or hyperactivity-motor impulsiveness. Neuropsychological evidence indicates the comorbidity of anxiety disorder with AD/HD. The aim of the present study was to characterize anxiety-related behavior of the juvenile stroke-prone spontaneously hypertensive rat (SHRSP), an animal model of AD/HD, and compare with genetic and/or normotensive controls. Our hypothesis, that low susceptibility to fear/anxiety stimuli might underlie impulsive behavior in juvenile SHRSP, was assessed by a contextually conditioned fear paradigm. In order to examine whether contextual fear stimuli would affect the synaptic efficacy in the hippocampus, changes in field potentials of perforant path-dentate gyrus (DG) synapses were recorded in the freely behaving rat. Aversive footshock (FS) stimuli elicited intense freezing behavior in genetic and/or normotensive controls. Re-exposing to the FS chamber in the 30-min retention period also produced freezing behavior, as contextually conditioned fear response, in genetic and/or normotensive controls. In contrast, SHRSP exhibited a significant attenuation in freezing behavior both in the 5-min post FS period (immediately after FS) and in the 30-min retention period (24 hours after FS) as compared to genetic and/or normotensive controls. Pain perception as measures of behavioral responses to electric FS, jumping and/or vocalization, indicated that less anxiety-related response in SHRSP did not simply result from low susceptibility to FS stimuli. During the re-exposure to contextual fear stimuli, SHRSP and a normotensive control rat exhibited a decrease in the amplitude of the evoked population spikes in perforant path-DG synapses, accompanying freezing behavior. The synaptic response in this hippocampal subfield was mimicked by low frequency stimulation (1 Hz). These synaptic responses induced by behavioral and electrophysiological manipulations, freezing and LFS, were less pronounced in SHRSP than those in the control. Our findings indicate that the impaired responsiveness to contextually conditioned fear stimuli, less anxiety-related freezing behavior, in juvenile SHRSP, might explain the impulsive behavior in this AD/HD animal model.

Learning and memory in agmatine-treated rats

Agmatine, a noncompetitive N-methyl- d-aspartate (NMDA) antagonist, was examined for its role in water maze place learning, contextual and auditory-cued (discrete) fear learning and conditioned taste aversion learning, when administered systemically. Male Wistar rats were given saline or 1, 5, 10 or 50 mg/kg agmatine ip 20 min prior to or 30 min following daily training sessions in a hidden-platform (place learning) water maze task. Agmatine did not affect latencies to find the hidden platform or preference for the training quadrant during probe trials. When administered 20 min prior to contextual or auditory-cued fear-conditioning sessions, these doses of agmatine evoked a linear dose-dependent impairment in the magnitude of learned fear to the contextual stimuli when assessed during extinction trials 24 h later, but had no effect on the magnitude of learned fear to the auditory stimulus. Inferences of baseline motor activity and ability to respond to the presentation of footshock stimuli were not affected by the treatment. Injections of 50 mg/kg agmatine concurrently with a malaise-evoking agent following presentations to a novel sucrose solution abolished learned taste aversions; this agent did not evoke conditioned taste aversions alone. These studies indicate that systemically administered agmatine selectively impairs behavioral inferences of specific types of learning and memory.

Emotional and footshock stimuli induce differential long-lasting behavioural effects in rats; involvement of opioids

Rats were exposed to either a footshock stimulus (FS) or emotional stimulus (ES, forced perception of another rat receiving footshocks) during a daily 10-min session for 5 consecutive days. The consequences of FS and ES on their behavioural responsiveness were assessed at different post-stress intervals using a small open-field. FS induced a decrease in ambulation, rearing and sniffing and an increased immobility in the small open field. These effects were present in rats tested immediately after the last session and remained present for at least 15 days. In contrast, ES induced a transient decrease in ambulation and rearing immediately after the last session, but in the period from half an hour until at least 15 days after the stimulus experience, an increase in ambulation, rearing and sniffing was observed. Exposure to one footshock per session for 5 consecutive days or to 10 footshocks in a single session also resulted in a long-lasting reduction in ambulation and sniffing and an increase in immobility. The former regime did not influence the behavioural response of ES rats, but the latter resulted in an increase in ambulation, rearing and sniffing in ES rats. Naloxone (1 mg/kg s.c.) pretreatment antagonized the increased behavioural activity of the ES rats whereas the activity of control and FS animals was not affected, suggesting an involvement of endogenous opioid systems in the behavioural responses observed in ES rats. It is suggested that the behavioural responses of the ES and FS animals are regulated by different mechanisms.

Long-latency responses of brain noradrenergic neurons to noxious stimuli are preferentially attenuated by intravenous morphine

The nucleus locus coeruleus (LC) has been strongly implicated in the processing of noxious stimuli. Consistent with this, previous studies have shown that spontaneous LC discharge is depressed by morphine. However, effects of morphine on evoked responses of LC neurons to noxious stimuli have not been systematically examined. We reported recently that responses to footshock stimuli in rat locus coeruleus neurons consist of an early (A-fiber mediated) component and a previously undescribed late (C-fiber mediated) component. In the present study, we administered analgesic doses of morphine (0.1, 0.5, or 1.0 mg/kg, i.v.) to determine the effect on A- and C-fiber components of footshock responses in LC neurons. Doses of 0.5 and 1.0 mg/kg significantly attenuated the C-fiber mediated response of LC neurons without affecting the A-fiber response component. Spontaneous LC discharge was reduced by administration of all doses of morphine. Both depressive effects of morphine were abolished by intravenous administration of naloxone. In contrast, local microinfusion of naloxone into the LC abolished the morphine-induced decrease of spontaneous discharge but did not prevent the depression of the C-fiber mediated footshock response by morphine. This indicates that the site of action for morphine's attenuation of the late LC response to footshock stimulation is outside of the LC. The results are consistent with the hypothesis that the late (C-fiber-mediated) footshock responses in locus coeruleus are involved in the processing of noxious stimuli and may contribute to anti-nociceptive mechanisms.

Effect of ovarian steroids on footshock avoidance learning and retention in female mice

Mice were trained to avoid footshock in a T-maze, with retention tested one week later. Adult male CD-1 mice made their first avoidance during acquisition after fewer trials than random cycling females and with less variability. Female mice in diestrus, when plasma levels of progesterone are low, learned to avoid footshock faster than females in estrus. Ovariectomized (OVX) mice learned in fewer trials than intact random cycling mice. Similar differences, though of a smaller magnitude, were found on the retention tests (i.e. males had better retention than females, mice in diestrus showed better retention 8 days later when in the same part of the estrous cycle than thse in estrus, and OVX mice had better retention than cycling females). OVX mice with estrogen implants learned faster than those with progesterone implants or progesterone plus estrogen implants. Hormonal status did not affect sensitivity to acoustic or footshock stimuli as measured by a startle reflex, nor did it affect activity. Pretraining administration of amphetamine, picrotoxin and strychnine attenuated the impairing effect of progesterone on acquisition. The possibility that progesterone may impair learning and to some extent, retention by facilitating the GABAergic activity and thereby reducing arousal level is discussed.

A novel long-latency response of locus coeruleus neurons to noxious stimuli: mediation by peripheral C-fibers.

1. Extracellular recordings were obtained from single presumed noradrenergic neurons in the nucleus locus coeruleus (LC) in response to percutaneous electrical foot shock (FS) stimulation in the rat. We employed long-duration stimulus pulses to examine the possible contribution of peripheral C-fibers to evoked activity in LC. 2. As in previous studies, 0.5-ms FS stimuli produced a short-latency excitatory response (onset 20.8 ms) followed by a prolonged postactivation inhibition. However, 2.0- or 5.0-ms FS stimuli yielded an additional late excitatory response. 3. The onset and termination latency of this late excitatory response were approximately 200 and 400 ms, respectively, after the FS. The conduction velocity for this late response (peripheral plus central pathway) was estimated to be < 1.2 m/s. 4. The percentages of LC neurons that exhibited a significant late response was 4% (3/85) with 0.5-ms stimuli, 53% (31/59) with 2.0-ms stimuli, and 73% (47/64) with 5.0-ms FS stimuli. 5. The average number of spikes evoked per FS stimulus (presented at 0.5 Hz) increased from the 1st to the 21st FS stimulus. This increased response occurred selectively in the late, not in the early, response. This result was interpreted to represent a "windup" phenomenon reflected at the level of LC. 6. Direct application of capsaicin onto the sciatic nerve reduced the average magnitude of the late response of LC neurons to 17% of control 42.5 min after application. The magnitude of the short-latency response components showed little or no change. 7. Averaged compound action potential (cAP) recordings from the sciatic nerve revealed that C-fiber responses were more consistently observed and much larger with 5.0-ms compared with 0.5-ms FS stimuli. 8. C-fiber cAPs were reduced or eliminated after application of capsaicin distal to the recording site with a time course similar to that of the late response of LC neurons, with little or no effect on A-fiber cAPs. 9. These data show that a previously undescribed long-latency response of LC neurons to noxious FS stimulation in rat results from C-fiber activation in the sciatic nerve. This late response may be involved in generating descending noradrenergic analgesia induced by electrical stimulation of the foot pad or other body regions. Future studies of the pharmacology of this late response to noxious stimuli in LC may aid in developing new therapies for the treatment of acute or chronic pain.

Somatosensory and auditory convergence in the lateral nucleus of the amygdala.

Previous studies have shown that the lateral nucleus of the amygdala (AL) is essential in auditory fear conditioning and that neurons in the AL respond to auditory stimuli. The goals of the present study were to determine whether neurons in the AL are also responsive to somatosensory stimuli and, if so, whether single neurons in the AL respond to both auditory and somatosensory stimulation. Single-unit activity was recorded in the AL in anesthetized rats during the presentation of acoustic (clicks) and somatosensory (footshock) stimuli. Neurons in the dorsal subdivision of the AL responded to both somatosensory and auditory stimuli, whereas neurons in the ventrolateral AL responded only to somatosensory stimuli and neurons in the ventromedial AL did not respond to either stimuli. These findings indicate that the dorsal AL is a site of auditory and somatosensory convergence and may therefore be a locus of convergence of conditioned and unconditioned stimuli in auditory fear conditioning.

Effect of ethimizole on instrumental learning in rats

The action of etimizol on the acquisition of the conditioned reflexes with different complexity and biological significance of reinforcement was studied. The acquisition was performed in a shuttle box and U-shaped maze using food and footshock stimuli. The time interval between administration of etimizol (3 mg/kg) and the onset of learning varied between 0.5 and 3 h in the several series. Etimizol did not facilitate the learning in rats whatever the time of administration and biological modality of reinforcement. It is suggested that the positive effect of etimizol on the memory is related to its influence on the consolidation stage.

Unconditioned stress-induced analgesia following exposure to brief footshock.

Four experiments examined the properties of unconditioned analgesia elicited by electric footshock stimuli using unconditioned stimulus (US) parameters typical of aversive conditioning paradigms. In all experiments, analgesia was inferred from the latency to paw lick in response to painful thermal stimulation in the hot-plate assay. In Experiment 1, rats exposed to a 1-s, 2-mA electric shock US showed significantly longer latencies to respond to painful thermal stimulation than nonshocked controls, whereas nonsignificant increases in response latencies were observed with 1-s shock USs of either 0.5 or 1.25 mA. In Experiment 2, rats exposed to a 2-mA electric shock US showed systematic increases in latencies to respond to painful thermal stimulation as the duration of the shock was varied between 0.5 and 2 s. Experiment 3 showed that this form of shock-induced analgesia was of short temporal duration. Specifically, significant increases in latencies to respond to painful thermal stimulation occurred 30 s but not 90 or 300 s following exposure to a 1-s, 2-mA shock US. Experiment 4 demonstrated that this form of analgesia was unaffected by pretreatment with the opiate receptor antagonist naloxone in dosages of 1, 5, 10, or 20 mg/kg. Finally, there was no evidence showing that environmental stimuli paired with shock presentations acquired the capacity to evoke analgesia as a conditioned response. The implications of shock-induced analgesia for the study of aversive conditioning and behavior are discussed.

Reversible alterations in hypothalamic uptake of biogenic amines after septal lesions

We investigated the time course of several behavioral and anatomical correlates of the septal rage syndrome in rats. After lesion of the septal nuclei, rats were tested on several measures of sensory responsiveness at 1, 5, 10, or 15 days of recovery. The animals were then killed and the high-affinity uptake of tritiated norepinephrine (NE), dopamine (DA), and serotonin (5-HT) were assessed in crude synaptosome preparations of hippocamal and hypothalamic tissue. Behaviorally, rats with septal lesions exhibited a persistant increase in reactivity to heat and open-field stimuli while the initial increase in responsiveness to tactile and footshock stimuli progressively decreased during the 15-day recovery interval. Synaptosomes prepared from hippocampus of septal-lesion rats exhibited a progressive decrease in the uptake of tritiated NE, DA, and 5-HT which appeared maximal by day 15. In contrast, an initial decrease observed in the hypothalamic uptake of all three biogenic amine compounds returned to control values by day 15. We conclude that the septal rage syndrome exhibits a significant degree of spontaneous remission with time and can be dissociated from sensory hyperreactivity in general. Several consequences of septal lesions on the functional activity of biogenic amine neurons are discussed in light of their ability to explain the reversible alterations in hypothalamic uptake.