Beatriz Duarte Palma

Differential effects of acute cold and footshock on the sleep of rats

Several studies have shown that 1 h of immobilisation stress during the rats active period results in rebound of paradoxical (PS) and slow wave sleep (SWS). Since the effects of stress on behaviour and physiological parameters vary according to the stimulus, the present study sought to examine the activation of the hypothalamic-pituitary-adrenal (HPA) axis and the sleep pattern of rats submitted to 1 h of footshock, immobilisation or cold, or 18 h of PS deprivation (PSD). Stress sessions began between 0900 and 0930 h. Immediately after the end of the stress session, or at the corresponding time for controls, animals were blood sampled for determination of ACTH and corticosterone (CORT) plasma levels. In Experiment 2, animals were implanted with electrodes for basal and post-stress polysomnographic recording (6 h long). The results showed that all stressors produced an activation of the HPA axis; however, footshock induced the largest ACTH levels, whereas cold resulted in the highest CORT levels. In regard to the sleep data, immobilisation and PSD led to a rebound of SWS (+16.87% and +9.37%, respectively) and PS (+42.45% and +55.25%, respectively). Immobilisation, however, induced an increased number of PS episodes, whereas PSD resulted in longer PS episodes. Cold stress produced an exclusive rebound of SWS (+14.23%) and footshock promoted sustained alertness during the animals resting period (+47.18%). These results indicate that different stimuli altered the sleep pattern in a distinct manner; and these alterations might be related to the state of the HPA axis activation.

Pituitary-adrenal axis and behavioural responses of maternally deprived juvenile rats to the open field.

Adult rats submitted to maternal deprivation (DEP) on post-natal day (pnd) 11 show smaller corticosterone (CORT) response to a saline injection than non-deprived (NDEP) rats, mainly at 30 days of age. In the present study we sought to investigate the pituitary-adrenal axis response of 30-day-old DEP rats to a 5-min open-field session, with lights and sound on. Hormone levels were assessed immediately or 20 min after the end of the stress, and were compared to basal levels. The immediate ACTH response of DEP females was the highest; this difference was no longer observed at 20 min, but hormone levels were still higher than basal. Both males and females showed an augmented CORT secretion immediately after the open field session, although only the response of NDEP females was higher than that of male counterparts. Moreover, DEP females showed a lower CORT response than their NDEP counterparts immediately after the stress. The CORT stress response remained equally elevated in males and females at the 20-min time point. Finally, DEP animals, regardless of the gender, ambulated more in the centre of the open field and displayed less grooming behaviour than NDEP pups, suggesting that DEP rats are less emotional than NDEP animals.

Sleep rebound in animals deprived of paradoxical sleep by the modified multiple platform method.

The objective of the present study was to assess the sleep rebound of animals exposed to the modified multiple platform method (MMPM), in which cage-mate rats were placed onto narrow platforms (NP=6.5 cm in diameter), onto wide platforms (WP=14 cm in diameter) or onto a grid (GR). The last two groups were included as environmental controls for the deprivation method. Animals were implanted with bipolar electrodes in the cortex, hippocampus and neck muscle. Baseline sleep was recorded for 6 h, after which the animals were placed in one of the above-mentioned settings for 90 h and their sleep was again recorded. Comparison between baseline and post-GR recordings revealed no sleep differences in these animals. Placement of animals onto WP resulted in augmented sleep time (16%), time spent in PS (+99%), duration of PS episodes (+77%), sleep efficiency (+16%), and in reduced latency to PS (-84.8%). Finally, NP animals exhibited a dramatic increase in sleep time (+34.3%), time spent in PS (+184.7%), duration of PS episodes (+106%), and in sleep efficiency (+34.4%). Moreover, sleep latency (-52.2%) and time spent in SWS (-12.2%) were reduced. Based on the results of sleep rebound, the data indicated that placement of animals onto narrow platforms in the MMPM was an effective PS deprivation method and the grid should be considered as an adequate environmental control.

Paradoxical sleep deprivation increases plasma endothelin levels.

The endothelins (ET-1, 2 and 3) constitute a family of 21 amino acid peptides with potent biological activities. ET-1 is one of the most potent endogenous vasoconstrictors so far identified and its increased concentration in plasma appears to be closely related to the pathogenesis of arterial hypertension as well as to obstructive sleep apnea (OSA). OSA patients exhibit repetitive episodes of apnea and hypopnea that result in hypoxia and consecutive arousals. These patients are chronically sleep deprived, which may aggravate the hypertensive features, since literature data show that sleep deprivation results in hypertension both in humans and in animals. Based on the reported relationship between ET-1, hypertension and sleep deprivation consequences, the purpose of the present study was to determine plasma ET concentrations in paradoxical sleep-deprived animals. Male Wistar rats, 3 to 4 months old (N = 10 per group), were deprived of sleep for 24 and 96 h by the platform technique and plasma ET-(1/2) was measured by radioimmunoassay. Analysis of plasma revealed that 96 h of sleep deprivation induced a significant increase in ET-(1/2) release (6.58 fmol/ml) compared to control (5.07 fmol/ml). These data show that sleep deprivation altered plasma ET-(1/2) concentrations, suggesting that such an increase may participate in the genesis of arterial hypertension and cardiorespiratory changes observed after sleep deprivation.

Repercussões imunológicas dos distúrbios do sono: o eixo hipotálamo-pituitária-adrenal como fator modulador

OBJECTIVE: To review the literature on the interaction between sleep and the immune system. METHOD: A search on Web of Science and Pubmed database including the keywords sleep, sleep deprivation, stress, hypothalamic-pituitary-adrenal axis, immune system, and autoimmune diseases. RESULTS: On Web of Science, 588 publications were retrieved; 61 references, more significant and closer to our objective, were used, including original articles and review papers. CONCLUSION: Sleep deprivation and immune system exert a bidirectional influence on each other. Since sleep deprivation is considered a stressor, inasmuch as it induces elevation of cortisol or corticosterone levels in humans and rodents, respectively, and given the well-known immunosuppressive effect of glucocorticoids, we propose that increased activation of the hypothalamic-pituitary-adrenal axis is a major mediator of the immune alterations observed in patients with insomnia or in sleep deprived subjects.

The effects of acute cocaine administration in paradoxical sleep‐deprived rats

Recent studies demonstrate the action of cocaine on reward pathways, which are activated by pleasant stimuli. Cocaines mechanism of action involves the blockade of dopamine and norepinephrine reuptake by the presynaptic terminal. Paradoxical sleep (PS) deprivation is known to induce several behavioural alterations most of which suggest the occurrence of supersensitivity of D 2 and a subsensitivity of b 1 and b 2 receptors.The present study sought to examine the effects of PS deprivation on the actions of cocaine on the erection and ejaculation behaviours in rats. Four different doses of cocaine (3.5, 7.0, 15.0 and 30.0 mg/kg) were acutely administered to Wistar male rats, at the end of a 4‐day period of PS deprivation or at the equivalent time‐point to control animals. Moreover, 15 mg/kg of cocaine were administered in animals submitted to immobilization, footshock and forced swimming. The data were analysed by the Chi‐square test, and revealed that only PS‐deprived animals exhibited penile erections and ejaculation behaviours, which were absent in control animals. PS deprivation increases the sexual behaviour of male rats, probably due to its action on dopaminergic systems. However, further studies need to be carried out in order to clarify the mechanisms involved between PS deprivation and cocaine.

Effects of early handling on basal and stress-induced sleep parameters in rats

Exposure of humans and animals to stressful events early in life leads to significant and often permanent behavioural, neuroendocrine and central alterations. Early handling consists of removing the litter from the nest for 15 min/day, from post-natal days 2 to 14 and results in lowered ACTH and corticosterone stress response and reduced anxiety-like and fear behaviours. Stress-induced sleep alterations usually consists of increased sleep time, known as sleep rebound. In the present study, basal and stress-induced sleep pattern of control non-manipulated (CTL) and early handled (EH) adult male rats was investigated. Sleep was evaluated by 21-h polysomnographic recordings (from 10:00 to 07:00 h of the next day) before and after a 1-h session of restraint stress. The results showed that in the first 3 h following stress, both CTL and EH animals exhibited an impairment of sleep, with a reduction of sleep efficiency, duration of slow wave sleep and of paradoxical sleep. On the contrary, time awake and awakening bouts were augmented in this period. Sleep rebound was observed mainly in the dark period of the light-dark cycle. Stress-induced sleep changes were similar between CTL and EH animals for most sleep parameters. However, EH animals exhibited more bouts of paradoxical sleep on the night following stress exposure and longer bouts of paradoxical sleep in the light period that followed restraint stress. These data indicate that stress-induced alterations of sleep in early handled animals are similar to that observed in control animals, except for some parameters related to paradoxical sleep.

Inflammatory markers are associated with inhibitory avoidance memory deficit induced by sleep deprivation in rats

Sleep deprivation (SD) causes detrimental effects to the body, such as memory impairment and weight loss. SD also changes the concentration of inflammatory mediators such as cytokines, which, in turn, can affect cognitive functioning. Thus, the objective of this study was to investigate the involvement of these inflammatory mediators in inhibitory avoidance memory deficit in sleep-deprived rats. Male Wistar rats were deprived of sleep by the modified multiple platform method for 96 h, while their respective controls remained in their housing cages. To assess memory after SD, all animals underwent training, followed by the inhibitory avoidance task test 24h later. Also, the weight of each animal was recorded daily. In the first experiment, animals received an acute administration of lipopolysaccharide (LPS, 50 or 75 μg/kg i.p.) 3h before the inhibitory avoidance training. In the experiment 2, the animals received acute or chronic administration of anti-IL-6 antibody (Ab, 2 μg/kg i.p.). The acute administration was performed 3h before the inhibitory avoidance training, while the chronic treatment administrations were performed daily during the SD period. The 75 μg/kg dose of LPS, but not the 50 μg/kg dose, caused a significant attenuation of memory impairment in the sleep-deprived animals. Although the treatments with the anti-IL-6 Ab did not produce any significant changes in cognitive performance, the Ab attenuated weight loss in sleep-deprived animals. Taken together, these results suggest the involvement of inflammatory mediators in the modulation of memory deficit and weight loss that are observed in sleep-deprived rats.

Effect of Sleep Deprivation on the Corticosterone Secretion in an Experimental Model of Autoimmune Disease

Objective: Sleep disturbances have been observed in a number of chronic inflammatory conditions, such as systemic lupus erythematosus. Previous results from our laboratory showed that when NZB/NZWF1 mice, an experimental model of lupus, are submitted to sleep deprivation (SD), they exhibit an earlier onset of the disease. Sleep disturbances have far-reaching effects on the endocrine and immune system, changes that may be linked to disease manifestation. Immunoendocrine communication via the hypothalamic-pituitary-adrenal axis has been proposed as an important modulatory factor for the development of autoimmune disease. We hypothesized here that corticosterone (CORT) could be involved in earlier onset of the disease in sleep-deprived NZB/NZWF1 mice. Methods: The profile of CORT secretion was measured immediately after the end of SD (platform method) and during the development of the disease. Also, we analyzed the effects of SD on CORT secretion of Swiss albino mice, which do not present immune alterations. Results: The results showed that NZB/NZWF1 mice exhibited a CORT response to SD similar to Swiss albino mice. However, CORT levels remained elevated throughout the whole period of evaluation. There was an increase in circulating levels of CORT in the NZB/NZWF1 mice as the disease progressed, but this effect was more evident in the sleep-deprived mice. Conclusion: According to these results, we suggest that elevated CORT levels are involved in the earlier onset of the disease.

Effects on prolactin secretion and binding to dopaminergic receptors in sleep-deprived lupus-prone mice

Sleep disturbances have far-reaching effects on the neuroendocrine and immune systems and may be linked to disease manifestation. Sleep deprivation can accelerate the onset of lupus in NZB/NZWF(1) mice, an animal model of severe systemic lupus erythematosus. High prolactin (PRL) concentrations are involved in the pathogenesis of systemic lupus erythematosus in human beings, as well as in NZB/NZWF(1) mice. We hypothesized that PRL could be involved in the earlier onset of the disease in sleep-deprived NZB/NZWF(1) mice. We also investigated its binding to dopaminergic receptors, since PRL secretion is mainly controlled by dopamine. Female NZB/NZWF(1) mice aged 9 weeks were deprived of sleep using the multiple platform method. Blood samples were taken for the determination of PRL concentrations and quantitative receptor autoradiography was used to map binding of the tritiated dopaminergic receptor ligands [3H]-SCH23390, [3H]-raclopride and [3H]-WIN35,428 to D(1) and D(2) dopaminergic receptors and dopamine transporter sites throughout the brain, respectively. Sleep deprivation induced a significant decrease in plasma PRL secretion (2.58 +/- 0.95 ng/mL) compared with the control group (25.25 +/- 9.18 ng/mL). The binding to D(1) and D(2) binding sites was not significantly affected by sleep deprivation; however, dopamine transporter binding was significantly increased in subdivisions of the caudate-putamen--posterior (16.52 +/- 0.5 vs 14.44 +/- 0.6), dorsolateral (18.84 +/- 0.7 vs 15.97 +/- 0.7) and ventrolateral (24.99 +/- 0.5 vs 22.54 +/- 0.7 microCi/g), in the sleep-deprived mice when compared to the control group. These results suggest that PRL is not the main mechanism involved in the earlier onset of the disease observed in sleep-deprived NZB/NZWF(1) mice and the reduction of PRL concentrations after sleep deprivation may be mediated by modifications in the dopamine transporter sites of the caudate-putamen.