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To evaluate the effect of weight loss induced by dietetic treatment, with or without an intragastric balloon, on gastric emptying of obese subjects.20 morbidly obese subjects (21-54 years, 45.3-58.0 kg/m2) and 20 healthy controls (21-56 years, 20.3-24.8 kg/m2).Parallel study of a 4 month, low calorie dietetic treatment with or without a 500 ml intragastric balloon.In basal conditions, obese subjects had accelerated gastric emptying as compared to healthy controls. At the end of the dietetic treatment period, a significant decrease of body weight was obtained. Patients also showed a slowing of gastric emptying. Both the weight loss and the slowing of gastric emptying occurred irrespective of the presence or absence of the intragastric balloon.The present findings are compatible with the hypothesis that gastric emptying, food intake and body weight are integrated parameters in subjects with morbid obesity.

Early-life exposure to environmental toxins like tobacco can permanently re-program body structure and function. Here, we investigated the long-term effects on mouse adult sleep phenotype exerted by early-life exposure to nicotine or to its principal metabolite, cotinine. Moreover, we investigated whether these effects occurred together with a reprogramming of the activity of the hippocampus, a key structure to coordinate the hormonal stress response. Adult male mice born from dams subjected to nicotine (NIC), cotinine (COT) or vehicle (CTRL) treatment in drinking water were implanted with electrodes for sleep recordings. NIC and COT mice spent significantly more time awake than CTRL mice at the transition between the rest (light) and the activity (dark) period. NIC and COT mice showed hippocampal glucocorticoid receptor (GR) downregulation compared to CTRL mice, and NIC mice also showed hippocampal mineralocorticoid receptor downregulation. Hippocampal GR expression significantly and inversely correlated with the amount of wakefulness at the light-to-dark transition, while no changes in DNA methylation were found. We demonstrated that early-life exposure to nicotine (and cotinine) concomitantly entails long-lasting reprogramming of hippocampal activity and sleep phenotype suggesting that the adult sleep phenotype may be modulated by events that occurred during that critical period of life.

The loss of hypothalamic neurons that produce wake-promoting orexin (hypocretin) neuropeptides is responsible for narcolepsy type 1 (NT1). While the number of histamine neurons is increased in patients with NT1, results on orexin-deficient mouse models of NT1 are inconsistent. On the other hand, the effect of histamine deficiency on orexin neuron number has never been tested on mammals, even though histamine has been reported to be essential for the development of a functional orexin system in zebrafish. The aim of this study was to test whether histamine neurons are increased in number in orexin-deficient mice and whether orexin neurons are decreased in number in histamine-deficient mice. The hypothalamic neurons expressing L-histidine decarboxylase (HDC), the histamine synthesis enzyme, and those expressing orexin A were counted in four orexin knock-out mice, four histamine-deficient HDC knock-out mice, and four wild-type C57BL/6J mice. The number of HDC-positive neurons was significantly higher in orexin knock-out than in wild-type mice (2,502 ± 77 vs. 1,800 ± 213, respectively, one-tailed t-test, P = 0.011). Conversely, the number of orexin neurons was not significantly lower in HDC knock-out than in wild-type mice (2,306 ± 56 vs. 2,320 ± 120, respectively, one-tailed t-test, P = 0.459). These data support the view that orexin peptide deficiency is sufficient to increase histamine neuron number, supporting the involvement of the histamine waking system in the pathophysiology of NT1. Conversely, these data do not support a significant role of histamine in orexin neuron development in mammals.

Antihistamine medications have been suggested to elicit clinical features of restless legs syndrome. The available data are limited, particularly concerning periodic leg movements during sleep, which are common in restless legs syndrome and involve bursts of tibialis anterior electromyogram. Here, we tested whether the occurrence of tibialis anterior electromyogram bursts during non-rapid eye movement sleep is altered in histidine decarboxylase knockout mice with congenital histamine deficiency compared with that in wild-type control mice. We implanted six histidine decarboxylase knockout and nine wild-type mice to record neck muscle electromyogram, bilateral tibialis anterior electromyogram, and electroencephalogram during the rest (light) period. The histidine decarboxylase knockout and wild-type mice did not differ significantly in terms of sleep architecture. In both histidine decarboxylase knockout and wild-type mice, the distribution of intervals between tibialis anterior electromyogram bursts had a single peak for intervals < 10s. The total occurrence rate of tibialis anterior electromyogram bursts during non-rapid eye movement sleep and the occurrence rate of the tibialis anterior electromyogram bursts separated by intervals < 10s were significantly lower in histidine decarboxylase knockout than in wild-type mice. These data do not support the hypothesis that preventing brain histamine signalling may promote restless legs syndrome. Rather, the data suggest that limb movements during sleep, including those separated by short intervals, are a manifestation of subcortical arousal requiring the integrity of brain histamine signalling.

Under conditions of scarce food availability and cool ambient temperature, the mouse (Mus Musculus) enters into torpor, a state of transient metabolic suppression mediated in part by the autonomic nervous system. Hypothalamic orexins are involved in the coordination of behaviors and autonomic function. We tested whether orexins are necessary for the coordinated changes in physiological variables, which underlie torpor and represent its physiological signature. We performed simultaneous measurements of brain temperature, electroencephalographic, and electromyographic activity allowing objective assessment of wake-sleep behavior, and cardiovascular, respiratory, and metabolic variables in orexin knockout mice (ORX-KO) and wild-type mice (WT) during torpor bouts elicited by caloric restriction and mild cold stress. We found that torpor bouts in WT are characterized by an exquisitely coordinated physiological signature. The characteristics of torpor bouts in terms of duration and rate of change of brain temperature and electromyographic activity at torpor entrance and exit did not differ significantly between ORX-KO and WT, and neither did the cardiovascular, respiratory, and metabolic characteristics of torpor. ORX-KO and WT also had similar wake-sleep state changes associated with torpor bouts, with the exception of a significantly higher rapid-eye movement sleep time in ORX-KO at torpor entrance. Our results demonstrate that orexins are not necessary either for the normal physiological adaptations occurring during torpor in mice or for their coordination, suggesting that mechanisms different from orexin peptide signaling may be involved in the regulation and the coordination of these physiological responses.

The integrated activity of the autonomic nervous system (ANS) with the somatomotor and the neuroendocrine systems allows an animal to maintain internal homoeostasis and regulate its interaction with the external environment. Significant changes in ANS activity occur on passing from wakefulness to sleep, and this pattern of autonomic activation is profoundly different even between NREMS and REMS. While, during NREMS, the ANS works to maintain body homeostasis in accordance with the reduced metabolic needs of this state, during REMS ANS activity is greatly variable, and not visibly oriented to the maintenance of the stability of physiological variables. Sleep derangements determine an alteration of autonomic activity that can also extend to waking, favoring the development of pathological conditions. In this review, we summarize new insights into the impact of sleep derangements on ANS control of immune, respiratory and cardiovascular function, and into the role of ANS in thermoregulation and energy expenditure.

Study objectives: The use of mouse models in sleep apnea study is limited by the belief that central (CSA) but not obstructive sleep apneas (OSA) occur in rodents. We aimed to develop a protocol to investigate the presence of OSAs in wild-type mice and, then, to apply it to a validated model of Down syndrome (Ts65Dn), a human pathology characterized by a high incidence of OSAs. Methods: In a pilot study, nine C57BL/6J wild-type mice were implanted with electrodes for electroencephalography (EEG), neck electromyography (nEMG), and diaphragmatic activity (DIA), and then placed in a whole-body-plethysmographic (WBP) chamber for 8 h during the rest (light) phase to simultaneously record sleep and breathing activity. CSA and OSA were discriminated on the basis of WBP and DIA signals recorded simultaneously. The same protocol was then applied to 12 Ts65Dn mice and 14 euploid controls. Results: OSAs represented about half of the apneic events recorded during rapid-eye-movement-sleep (REMS) in each experimental group, while the majority of CSAs were found during non-rapid eye movement sleep. Compared with euploid controls, Ts65Dn mice had a similar total occurrence rate of apneic events during sleep, but a significantly higher occurrence rate of OSAs during REMS, and a significantly lower occurrence rate of CSAs during NREMS. Conclusions: Mice physiologically exhibit both CSAs and OSAs. The latter appear almost exclusively during REMS, and are highly prevalent in Ts65Dn. Mice may, thus, represent a useful model to accelerate the understanding of the pathophysiology and genetics of sleep-disordered breathing and to help the development of new therapies.

The loss of orexinergic neurons, releasing orexins, results in narcolepsy. Orexins participate in the regulation of many physiological functions, and their role as wake-promoting molecules has been widely described. Less is known about the involvement of orexins in body temperature and respiratory regulation. The aim of this study was to investigate whether orexin peptides modulate respiratory regulation as a function of ambient temperature (T°a) during different sleep stages. Respiratory phenotype of male orexin knockout (KO-ORX, n=9) and wild-type (WT, n=8) mice was studied at thermoneutrality (T°a=30°C) or during mild cold exposure (T°a=20°C) inside a whole-body plethysmography chamber. The states of wakefulness (W), non-rapid-eye-movement sleep (NREMS) and rapid-eye-movement sleep (REMS) were scored non-invasively, using a previously validated technique. Both in WT and KO-ORX mice T°a strongly and significantly affected ventilatory period and minute ventilation values during NREMS and REMS; moreover, the occurrence rate of sleep apneas in NREMS was significantly reduced at T°a=20°C compared to T°a=30°C. Overall, there were no differences in respiratory regulation during sleep between WT and KO-ORX mice, except for sigh occurrence rate, which was significantly increased at T°a=20°C with respect to T°a =30°C in WT mice, but not in KO-ORX mice. These results do not support a main role for orexin peptides in the temperature-dependent modulation of respiratory regulation during sleep. However, we showed that the occurrence rate of sleep apneas critically depends on T°a, without any significant effect of orexin peptides.

Sleep apneas can be categorized as post-sigh (prevailing in non-rapid eye movement sleep) or spontaneous (prevailing in rapid eye movement sleep) according to whether or not they are preceded by an augmented breath (sigh). Notably, the occurrence of these apnea subtypes changes differently in hypoxic/hypercapnic environments and in some genetic diseases, highlighting the importance of an objective discrimination. We aim to: (a) systematically review the literature comparing the criteria used in categorizing mouse sleep apneas; and (b) provide data-driven criteria for this categorization, with the final goal of reducing experimental variability in future studies. Twenty-two wild-type mice, instrumented with electroencephalographic/electromyographic electrodes, were placed inside a whole-body plethysmographic chamber to quantify sleep apneas and sighs. Wake-sleep states were scored on 4-s epochs based on electroencephalographic/electromyographic signals. Literature revision showed that highly different criteria were used for post-sigh apnea definition, the intervals for apnea occurrence after sigh ranging from 1 breath up to 20 s. In our data, the apnea occurrence rate during non-rapid eye movement sleep was significantly higher than that calculated before the sigh only in the 1st and 2nd 4-s epochs following a sigh. These data suggest that, in mice, apneas should be categorized as post-sigh only if they start within 8 s from a sigh; the choice of shorter or longer time windows might underestimate or slightly overestimate their occurrence rate, respectively.

Torpor is a peculiar mammalian behaviour, characterized by the active reduction of metabolic rate, followed by a drop in body temperature. To enter torpor, the activation of all thermogenic organs that could potentially defend body temperature must be prevented. Most of these organs, such as the brown adipose tissue, are controlled by the key thermoregulatory region of the Raphe Pallidus (RPa). Currently, it is not known which brain areas mediate the entrance into torpor. To identify these areas, the expression of the early gene c-Fos at torpor onset was assessed in different brain regions in mice injected with a retrograde tracer (Cholera Toxin subunit b, CTb) into the RPa region. The results show a network of hypothalamic neurons that are specifically activated at torpor onset and a direct torpor-specific projection from the Dorsomedial Hypothalamus to the RPa that could putatively mediate the suppression of thermogenesis during torpor.

Many small mammals, such as the laboratory mouse, utilize the hypometabolic state of torpor in response to caloric restriction. The signals that relay the lack of fuel to initiate a bout of torpor are not known. Because the mouse will only enter a torpid state when calorically challenged, it may be that one of the inputs for initiation into a bout of torpor is the lack of the primary fuel (glucose) used to power brain metabolism in the mouse. Using glucose telemetry in mice, we tested the hypotheses that 1) circulating glucose (GLC), core body temperature (Tb), and activity are significantly interrelated; and 2) that the level of GLC at the onset of torpor differs from both GLC during arousal from torpor and during feeding when there is no torpor. To test these hypotheses, six C57Bl/6J mice were implanted with glucose telemeters and exposed to different feeding conditions (ad libitum, fasting, limited food intake, and refeeding) to create different levels of GLC and Tb. We found a strong positive and linear correlation between GLC and Tb during ad libitum feeding. Furthermore, mice that were calorically restricted entered torpor bouts readily. GLC was low during torpor entry but did not drop precipitously as Tb did at the onset of a torpor bout. GLC significantly increased during arousal from torpor, indicating the presence of endogenous glucose production. While low GLC itself was not predictive of a bout of torpor, hyperactivity and low GLC preceded the onset of torpor, suggesting that this may be involved in triggering torpor.

Quest’opera illustra i fondamenti di anatomia e fisiologia del sistema nervoso centrale alla base dei comportamenti e delle funzioni psichiche nell’uomo ed è frutto del lavoro di un gruppo di eminenti esperti di neuroscienze. Per facilitare l’apprendimento, il testo è stato corredato da numerose illustrazioni e da materiali di approfondimento on line tra cui un tutorial per l’apprendimento dell’anatomia attraverso il disegno e sezioni di autoverifica. All’interno delle trattazioni sistematiche dei diversi argomenti si trovano inserti che illustrano metodiche di studio del sistema nervoso impiegate dalle moderne neuroscienze e conseguenze delle patologie neurologiche sul funzionamento del sistema nervoso.

Quest’opera illustra i fondamenti di anatomia e fisiologia del sistema nervoso centrale alla base dei comportamenti e delle funzioni psichiche nell’uomo ed è frutto del lavoro di un gruppo di eminenti esperti di neuroscienze. Per facilitare l’apprendimento, il testo è stato corredato da numerose illustrazioni e da materiali di approfondimento on line tra cui un tutorial per l’apprendimento dell’anatomia attraverso il disegno e sezioni di autoverifica. All’interno delle trattazioni sistematiche dei diversi argomenti si trovano inserti che illustrano metodiche di studio del sistema nervoso impiegate dalle moderne neuroscienze e conseguenze delle patologie neurologiche sul funzionamento del sistema nervoso.

Restless legs syndrome (RLS) is a neurological disorder that entails an urge to move with a circadian pattern during the evening/night. RLS may be accompanied by decreased sleep time and increased occurrence of periodic leg movements during sleep (PLMS), which involve bursts of tibialis anterior (TA) muscle electromyogram (EMG). Mild hypoxia and non-anemic iron deficiency, a highly prevalent nutritional deficiency, are relatively unexplored factors in RLS pathophysiology. We tested whether mice exposed to mild hypoxia, alone or in combination with non-anemic iron deficiency, show decreased sleep time particularly in the light (rest) period and increased occurrence of TA EMG phasic events similar to human PLMS. Female C57BL/6J mice were fed diets with low or normal iron for 6 months from weaning and instrumented with electrodes to record the electroencephalogram and the EMG of both TA muscles. Mice were recorded in a whole-body plethysmograph while breathing a normoxic or mildly hypoxic (15% O2) gas mixture for 48 h. Hypoxia increased minute ventilation during sleep. The low-iron diet decreased liver and serum iron, leaving blood hemoglobin and brainstem iron levels unaffected. Hypoxia, either alone or in combination with non-anemic iron deficiency, decreased non-rapid-eye-movement (non-REM) sleep time, but this occurred irrespective of the light/dark period and was not associated with increased occurrence of TA EMG events during non-REM sleep. These results do not support the hypothesis that mild hypoxia is sufficient to cause signs of RLS, either alone or in combination with non-anemic iron deficiency, pointing to the necessity of further susceptibility factors.

The validation of rodent models for restless legs syndrome (Willis-Ekbom disease) and periodic limb movements during sleep requires knowledge of physiological limb motor activity during sleep in rodents. This study aimed to determine the physiological time structure of tibialis anterior activity during sleep in mice and rats, and compare it with that of healthy humans. Wild-type mice (n = 9) and rats (n = 8) were instrumented with electrodes for recording the electroencephalogram and electromyogram of neck muscles and both tibialis anterior muscles. Healthy human subjects (31 ± 1 years, n = 21) underwent overnight polysomnography. An algorithm for automatic scoring of tibialis anterior electromyogram events of mice and rats during non-rapid eye movement sleep was developed and validated. Visual scoring assisted by this algorithm had inter-rater sensitivity of 92-95% and false-positive rates of 13-19% in mice and rats. The distribution of the time intervals between consecutive tibialis anterior electromyogram events during non-rapid eye movement sleep had a single peak extending up to 10 s in mice, rats and human subjects. The tibialis anterior electromyogram events separated by intervals

Background Scoring of wake–sleep states by trained investigators is a time-consuming task in many sleep experiments. We aimed to validate SCOPRISM, a new open-source algorithm for sleep scoring based on automatic graphical clustering of epoch distribution. Methods We recorded sleep and blood pressure signals of 36 orexin-deficient, 7 leptin knock-out, and 43 wild-type control mice in the PRISM laboratory. Additional groups of mice ( n = 14) and rats ( n = 6) recorded in independent labs were used to validate the algorithm across laboratories. Results The overall accuracy, specificity and sensitivity values of SCOPRISM (97%, 95%, and 94%, respectively) on PRISM lab data were similar to those calculated between human scorers (98%, 98%, and 94%, respectively). Using SCOPRISM, we replicated the main sleep and sleep-dependent cardiovascular findings of our previous studies. Finally, the cross-laboratory analyses showed that the SCOPRISM algorithm performed well on mouse and rat data. Comparison with existing methods SCOPRISM performed similarly or even better than recently reported algorithms. SCOPRISM is a very simple algorithm, extensively (cross)validated and with the possibility to evaluate its efficacy following a quick and easy visual flow chart. Conclusions We validated SCOPRISM, a new, automated and open-source algorithm for sleep scoring on a large population of mice, including different mutant strains and on subgroups of mice and rats recorded by independent labs. This algorithm should help accelerate basic research on sleep and integrative physiology in rodents.

"Neuroscienze cognitive. L’essenziale" introduce il lettore all'esplorazione scientifica della mente e del cervello approfondendo i meccanismi neurofisiologici sottostanti alle principali funzioni cognitive. Nonostante l’importanza che le neuroscienze cognitive attribuiscono alle basi biologiche dei fenomeni mentali, l’approccio utilizzato dall'autore non è la semplice descrizione dello stato dell’arte sull'argomento ma un percorso culturale finalizzato a rispondere alle perduranti domande su come l’esperienza, il pensiero e il comportamento nascono dalle attività cerebrali. Lo studente partecipa quindi attivamente al processo della scoperta scientifica. Il volume guida gli studenti e i dottorandi attraverso i temi e i principi fondamentali della disciplina in modo sintetico, organizzato e avvincente. Il testo è diviso in quattro sezioni: la prima, introduttiva, dedicata alla biologia del pensiero; la seconda focalizzata su sensazione, percezione, attenzione, e azione; la terza incentrata sulla rappresentazione mentale; la quarta che descrive le facoltà cognitive superiori.

The central neural mechanisms underlying differences in cardiovascular variability between wakefulness, non-rapid-eye-movement sleep (NREMS), and rapid-eye-movement sleep (REMS) remain poorly understood. These mechanisms may involve hypocretin (HCRT)/orexin signaling. HCRT signaling is linked to wake-sleep states, involved in central autonomic control, and impaired in narcoleptic patients. Thus, we investigated whether HCRT signaling plays a role in controlling cardiovascular variability during spontaneous behavior in HCRT-deficient mice. HCRT-ataxin3 transgenic mice lacking HCRT neurons (TG), knockout mice lacking HCRT peptides (KO), and wild-type controls (WT) were instrumented with electrodes for sleep recordings and a telemetric blood pressure transducer. Fluctuations of systolic blood pressure (SBP) and heart period (HP) during undisturbed wake-sleep behavior were analyzed with the sequence technique, cross-correlation functions, and coherent averaging of SBP surges. During NREMS, all mice had lower SBP variability, greater baroreflex contribution to HP control at low frequencies, and greater amplitude of the central autonomic and baroreflex changes in HP associated with SBP surges than during wakefulness. During REMS, all mice had higher SBP variability and depressed central autonomic and baroreflex HP controls relative to NREMS. HP variability during REMS was higher than during NREMS in WT only. TG and KO also had lower amplitude of the cardiac baroreflex response to SBP surges during REMS than WT. These results indicate that chronic lack of HCRT signaling may cause subtle alterations in the control of HP during spontaneous behavior. Conversely, the integrity of HCRT signaling is not necessary for the occurrence of physiological sleep-dependent changes in SBP variability.

Narcolepsy is a clinical condition characterized mainly by excessive sleepiness and cataplexy. Hypnagogic hallucinations and sleep paralysis complete the narcoleptic tetrad; disrupted night sleep, automatic behaviors and weight gain are also usual complaints. Different studies focus on autonomic changes or dysfunctions among narcoleptic patients, such as pupillary abnormalities, fainting spells, erectile dysfunction, night sweats, gastric problems, low body temperature, systemic hypotension, dry mouth, heart palpitations, headache and extremities dysthermia. Even if many studies lack sufficient standardization or their results have not been replicated, a non-secondary involvement of the autonomic nervous system in narcolepsy is strongly suggested, mainly by metabolic and cardiovascular findings. Furthermore, the recent discovery of a high risk for overweight and for metabolic syndrome in narcoleptic patients represents an important warning for clinicians in order to monitor and follow them up for their autonomic functions. We review here studies on autonomic functions and clinical disturbances in narcoleptic patients, trying to shed light on the possible contribute of alterations of the hypocretin system in autonomic pathophysiology.

In Spontaneously Hypertensive (SHR) and Wistar-Kyoto (WKY) normotensive rats, we quantified the extent to which spontaneous fluctuations of heart period (HP) may be determined from arterial pressure based on linear baroreflex properties. We analyzed time series (30-s length) of low-frequency (

We investigated whether the relative contribution of the baroreflex and central commands to the control of heart period differs between spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive rats (WKY) during physiological behavior. Rats were instrumented with an arterial catheter and with electrodes for discriminating wakefulness, nonrapid eye movement sleep (NREMS), and rapid eye movement sleep (REMS). The cross-correlation function (CCF) between spontaneous fluctuations of heart period and mean arterial pressure was computed at frequencies

During rapid-eye-movement (REM) sleep in adult subjects, the cerebral metabolic rate of oxygen consumption (CMRO(2)) is as high as that during wakefulness. We investigated whether CMRO(2) during active sleep is already at the waking level in newborn life, to support the role of active sleep as a state of endogenous brain activation during early postnatal development. Newborn lambs, 2-5 days old (n = 6), were instrumented with electrodes for sleep-state scoring, catheters for blood sample withdrawal and pressure monitoring, and a transit-time ultrasonic blood-flow probe around the superior sagittal sinus. At the age of 19 +/- 3 days, blood samples were obtained simultaneously from the carotid artery and the superior sagittal sinus during uninterrupted epochs of wakefulness, quiet sleep, and active sleep. The arteriovenous difference in blood oxygen concentration was multiplied by cerebral blood flow to determine CMRO(2). CMRO(2) during active sleep (47 +/- 5 micromol min(-1)) was similar to the value in wakefulness (44 +/- 6 micromol min(-1)) and significantly higher than in quiet sleep (39 +/- 5 micromol min(-1), P < 0.05). These data show that active sleep provides newborn lambs with brain activity at a level similar to that in wakefulness in terms of cerebral oxygen metabolism. The high CMRO(2) during active sleep supports its functional role during early postnatal development, when time spent in active sleep is at a lifetime maximum, albeit constituting a metabolic challenge for newborns, because of the impairment of systemic and cerebral vascular regulation in this sleep state.

Summary Hypocretin/orexin peptides are known for their role in the control of the wake–sleep cycle and narcolepsy–cataplexy pathophysiology. Recent studies suggested that hypocretin peptides also have a role in pregnancy. We tested this hypothesis by conducting a retrospective analysis on pregnancy complications in two different mouse models of hypocretin deficiency. We recorded 85 pregnancies of mice lacking either hypocretin peptides (knockout) or hypocretin-releasing neurons (transgenic) and their wild-type controls. Pregnancy was associated with unexplained dam death before delivery in 3/15 pregnancies in knockout mice, and in 3/23 pregnancies in transgenic mice. No casualties occurred in wild-type pregnant dams (P

Cannabinoid type 1 (CB1) receptors are highly expressed in the brain and play a role in behavior control. Endogenous cannabinoid signaling is modulated by high-fat diet (HFD). We investigated the consequences of congenital lack of CB1 receptors on sleep in mice fed standard diet (SD) and HFD. CB1 cannabinoid receptor knock-out (KO) and wild-type (WT) mice were fed SD or HFD for 4 months (n = 9-10 per group). Mice were instrumented with electroencephalographic (EEG) and electromyographic electrodes. Recordings were performed during baseline (48 hours), sleep deprivation (gentle handling, 6 hours), sleep recovery (18 hours), and after cage switch (insomnia model paradigm, 6 hours). We found multiple significant effects of genotype on sleep. In particular, KO spent more time awake and less time in non-rapid-eye-movement sleep (NREMS) and rapid-eye-movement sleep (REMS) than WT during the dark (active) period but not during the light (rest) period, enhancing the day-night variation of wake-sleep amounts. KO had slower EEG theta rhythm during REMS. REMS homeostasis after sleep deprivation was less effective in KO than in WT. Finally, KO habituated more rapidly to the arousing effect of the cage-switch test than WT. We did not find any significant effects of diet or of diet x genotype interaction on sleep. The occurrence of multiple sleep alterations in KO indicates important roles of CB1 cannabinoid receptors in limiting arousal during the active period of the day, in sleep regulation, and in sleep EEG in mice.

Body homeostasis and sleep homeostasis may both rely on the complex integrative activity carried out by the hypothalamus. Thus, the three main wake-sleep (WS) states (i.e. wakefulness, NREM sleep, and REM sleep) may be better understood if the different cardio-respiratory and metabolic parameters, which are under the integrated control of the autonomic and the endocrine systems, are studied during sleep monitoring. According to this view, many physiological events can be considered as an expression of the activity that physiological regulations should perform in order to cope with the need to fulfill body and sleep homeostasis. This review is aimed at making an assessment of data showing the existence of a physiological interplay between body homeostasis and sleep homeostasis, starting from the spontaneous changes observed in the somatic and autonomic activity during sleep, through evidence showing the deep changes occurring in the central integration of bodily functions during the different WS states, to the changes in the WS states observed when body homeostasis is challenged by the external environment and when the return to normal ambient conditions allows sleep homeo- stasis to run without apparent physiological restrictions. The data summarized in this review suggest that an approach to the dichotomy between NREM and REM sleep based on physiological regulations may offer a framework within which observations that a traditional behavioral approach may overlook can be interpreted. The study of the interplay between body and sleep homeostasis appears, therefore, to be a way to understand the function of complex organisms beyond that of the specific regulations.

Cannabinoid type 1 (CB1) receptors are expressed in the nervous and cardiovascular systems. In mice, CB1 receptor deficiency protects from metabolic consequences of a high-fat diet (HFD), increases sympathetic activity to brown fat, and entails sleep anomalies. We investigated whether sleep-wake and diet-dependent cardiorespiratory control is altered in mice lacking CB1 receptors. CB1 receptor knock-out (KO) and intact wild-type (WT) mice were fed standard diet or a HFD for 3 months, and implanted with a telemetric arterial pressure transducer and electrodes for sleep scoring. Sleep state was assessed together with arterial pressure and heart rate (home cage), or breathing (whole-body plethysmograph). Increases in arterial pressure and heart rate on passing from the light (rest) to the dark (activity) period in the KO were significantly enhanced compared with the WT. These increases were unaffected by cardiac (β1) or vascular (α1) adrenergic blockade. The breathing rhythm of the KO during sleep was also more irregular than that of the WT. A HFD increased heart rate, impaired cardiac vagal modulation, and blunted the central autonomic cardiac control during sleep. A HFD also decreased cardiac baroreflex sensitivity in the KO but not in the WT. In conclusion, we performed the first systematic study of cardiovascular function in CB1 receptor deficient mice during spontaneous wake-sleep behavior, and demonstrated that CB1 receptor KO alters cardiorespiratory control particularly in the presence of a HFD. The CB1 receptor signaling may thus play a role in physiological cardiorespiratory regulation and protect from some adverse cardiovascular consequences of a HFD.

The central neural pathways underlying the physiological coordination between thermoregulation and the controls of the wake-sleep behavior and cardiovascular function remain insufficiently understood. Growing evidence supports the involvement of hypocretin (orexin) peptides in behavioral, cardiovascular, and thermoregulatory functions. We investigated whether the effects of ambient temperature on wake-sleep behavior and cardiovascular control depend on the hypothalamic neurons that release hypocretin peptides. Orexin-ataxin3 transgenic mice with genetic ablation of hypocretin neurons (n = 11) and wild-type controls (n = 12) were instrumented with electrodes for sleep scoring and a telemetric blood pressure transducer. Simultaneous sleep and blood pressure recordings were performed on freely-behaving mice at ambient temperatures ranging between mild cold (20°C) and the thermoneutral zone (30°C). In both mouse groups, the time spent awake and blood pressure were higher at 20°C than at 30°C. The cold-related increase in blood pressure was significantly smaller in rapid-eye-movement sleep (REMS) than either in non-rapid-eye-movement sleep (NREMS) or wakefulness. Blood pressure was higher in wakefulness than either in NREMS or REMS at both ambient temperatures. This effect was significantly blunted in orexin-ataxin3 mice irrespective of ambient temperature and particularly during REMS. These data demonstrate that hypocretin neurons are not a necessary part of the central pathways that coordinate thermoregulation with wake-sleep behavior and cardiovascular control. Data also support the hypothesis that hypocretin neurons modulate changes in blood pressure between wakefulness and the sleep states. These concepts may have clinical implications in patients with narcolepsy with cataplexy, who lack hypocretin neurons.

A daily rhythm of blood pressure (BP), with maximum values in the activity period, carries important prognostic information. The extent to which this rhythm depends on behavioral factors remains debated. Mice are the species of choice for functional genomics. In mice, episodes of wakefulness and sleep are not restricted to particular daily periods, allowing BP in each wake-sleep state to be measured at each time of day. The aim of this study was to investigate whether a circadian rhythm of BP is manifest in each wake-sleep state in mice. Mice with B6 genetic background (n = 26) were implanted with a telemetric BP transducer and electrodes to discriminate wake-sleep states and recorded while housed under a 12:12 h light-dark period. For each mouse, 8 values of BP were obtained in each wake-sleep state (wakefulness, non-rapid-eye-movement sleep, and rapid-eye-movement sleep) by averaging over successive 3-h time bins. Analysis of variance evidenced a significant time effect in each wake-sleep state as well as a significant wake-sleep state × time interaction effect. In an additional group of mice (n = 3) recorded in constant darkness, the Lomb-Scargle periodogram also revealed a significant circadian rhythm of BP in each wake-sleep state. These findings demonstrate that during each wake-sleep state, mice show daily and circadian rhythms of BP in conditions of entrainment to the light-dark cycle and in free-running conditions of constant darkness, respectively.

Neurons that release hypocretin (HCRT; orexin) peptides control wake–sleep states and autonomic functions, and are lost in patients with narcolepsy with cataplexy. Bursts of high-amplitude electroencephalographic (EEG) activity have been reported during behavioural arrests and rapid eye movement sleep (REMS) episodes at sleep onset in HCRT-deficient narcoleptic mice. Quantitative information on these EEG phenomena is lacking. We aimed to quantify EEG frequency, occurrence rate, daily rhythm and cardiovascular correlates of high-amplitude EEG bursts during REMS and cataplexy. Twenty HCRT-deficient mice and 15 congenic wild-type controls were instrumented with electrodes for sleep recordings and a telemetric blood pressure transducer. Short (1–2 s) high-amplitude bursts of pointed theta waves (7 Hz) occurred during either REMS or cataplexy in 80% of HCRT-deficient mice without any significant accompanying modification in systolic blood pressure or heart period. Theta bursts were significantly more likely to occur during the dark period and in the last third of REMS episodes. Similar EEG events were detected in a significantly lower fraction (27%) of wild-type mice and with a significantly lower occurrence rate (0.8 versus 5 per hour of REMS). These data demonstrate that occurrence of high-amplitude theta bursts is facilitated during REMS and cataplexy in narcoleptic mice. Analysis of EEG frequency and daily and intra-episode patterns of event occurrence do not support interpretation of theta bursts as temporally displaced pre-REMS spindles. Facilitation of high-amplitude theta bursts may thus represent a novel neurophysiological abnormality associated with chronic HCRT deficiency.

Hypothalamic neurons releasing hypocretin (HCRT) control wake-sleep behavior, cardiovascular system and body temperature and their loss entails narcolepsy. We investigated whether HCRT neurons mediate sleep-dependent cardiovascular adaptations to changes in ambient temperature (Ta). Narcoleptic mice with genetic ablation of HCRT neurons (n=11) and wild-type controls (n=12) were implanted with a telemetric blood pressure transducer and electrodes to discriminate wake-sleep behavior. Recordings were performed in each mouse at ambient temperatures of 25°C (acclimation temperature), 30°C, and 20°C in random order. Mean blood pressure (MBP) and heart rate (HR) were computed in each wake-sleep behavior and analyzed with 3-way analysis of variance and t-tests (significance at P < 0.05). Results revealed a significant interaction between the wake-sleep behavior and Ta on MBP and HR in both mouse strains, with MBP and HR rising at 20 °C, particularly during wakefulness and non-rapid-eye-movement sleep. No effect involving the mouse strain was statistically significant. These results demonstrate that hypothalamic HCRT neurons are not necessary for sleep-dependent cardiovascular adaptations to changes in Ta.

This Chapter is aimed to provide a general overview of the hypothalamus and its functions. The main features of hypothalamic functional neuroanatomy will be addressed. This will lay the ground to discuss the role of hypothalamic neural pathways in different body functions, such as the regulation of wake-sleep behavior, body temperature, body fluid osmolarity, energy balance, and the cardiovascular system. In this picture, the role of hypothalamic hypocretin neurons in orchestrating behavioral and autonomic responses to environmental challenges will stand out. Knowledge of such integrative physiologic role of hypocretin neurons is a prerequisite to understand the pathophysiology of patients with narcolepsy-cataplexy, in whom hypocretin neurons are functionally lost.

Blood pressure (BP) is higher in the activity than in the rest period. This daily BP rhythm has prognostic significance and is mainly attributed to behavioral factors, BP being higher in wakefulness than in sleep. A circadian BP rhythm independent of sleep is debated. Wake-sleep (W-S) episodes occur throughout the 24 hour day in rodents, allowing BP measurements in each W-S state at each time of day. Aim of this study was to investigate whether circadian BP rhythms occur in each W-S state in mice, which are the species of choice for functional genomics. Mice with B6 genetic background (n=26) were implanted with a telemetric BP transducer and electrodes to discriminate W-S states and recorded with 12:12 hours light-dark period. Mean BP values in wakefulness were averaged over 3-hour time bins and analyzed as z-scores. The same procedure was performed in non-rapid eye movement sleep and rapid eye movement (REM) sleep. Analysis of variance evidenced a significant time effect on BP in each W-S state as well as a significant W-S state x time interaction effect. The standard deviation of BP values among time bins was the highest in REM sleep. In an additional group of mice (n=3) recorded for 7 days in constant darkness, Lomb-Scargle periodogram demonstrated significant circadian rhythms of BP in each W-S state in free-running conditions. These findings provide evidence that circadian rhythms of BP occur during each W-S state in mice.

Hypothalamic hypocretin (HCRT) neurons modulate wake-sleep behavior and autonomic functions, and their loss entails narcolepsy. In HCRT-deficient narcoleptic mice, high-amplitude and phasic electroencephalographic (EEG) bursts have been described during rapid-eye-movement sleep (REMS) and cataplexy and interpreted as temporally displaced sleep spindles. We tested this interpretation by performing a detailed quantitative analysis of these phasic EEG events. Twenty HCRT-deficient mice and 15 wild-type controls were implanted with EEG and electromyographic electrodes and a telemetric blood pressure transducer. Data were analyzed with Chi-Square, Mann-Whitney e Wilcoxon tests and significance at P < 0.05. Short (1-2 s) high-amplitude bursts of pointed theta waves (7 Hz) occurred during either REMS or cataplexy in 80% of HCRT-deficient mice. Similar EEG events were detected in a significantly lower fraction (27%) of wild-type mice and with a significantly lower occurrence rate (0.8 vs. 5 per hour of REMS). Theta bursts were significantly more likely to occur during the dark period and in the last third of REMS episodes, and were not associated with any significant cardiovascular change. The frequency and pattern of occurrence of theta bursts are not compatible with sleep spindles. Thus, theta wave bursts may represent a novel neurophysiological abnormality associated with HCRT deficiency.

With the onset to air-breathing at birth, the newborn circulation and its neural control undergoes dramatic transformation from the fetal pattern, yet remains substantially distinct from and less mature than the adult circulation. The newborn (particularly the preterm newborn) faces a risk of hypoxic-ischemic injury to vital organs as its blood pressure is at lifetime minimum, and mal-adaptation of the respiratory system leading to hypoxaemia is common. The brain is particularly vulnerable as cerebral blood flow is also at a lifetime minimum at birth. Coincidentally, REM sleep time is at a maximum in the newborn period. Organ perfusion may be problematic in REM sleep as central autonomic commands can induce blood pressure variations, while protective respiratory, circulatory and arousal reflexes can be ineffective. Local regulation of blood flow becomes critical if reflex control becomes ineffective. However this too may be ineffective for ensuring cerebral perfusion in REM sleep as cerebral autoregulation is slow in this sleep state compared to NREM sleep and wakefulness. Key factors modifying the cerebral autoregulatory curve are vasoconstrictor effects arising from cerebral sympathetic nerve activity (SNA), and counterbalancing vasodilator actions of endothelial-derived factors including nitric oxide (NO). Both SNA vasoconstriction and NO vasodilatation are prominently expressed in the newborn, particularly during REM sleep. Recent work suggests that each of these counterbalancing factors may normally be protective of the newborn brain (SNA through limiting microcirculatory pressure, flow and distension; NO by promoting perfusion). With further growth of the newborn in the neonatal and infant periods there is significant adaptation of the circulation and its autonomic neural control. Notably, and perhaps significant for the pathogenesis of sudden infant death syndrome (SIDS), adaptation occurs in postnatal age-related phases, rather than progressively with age. Moreover, this period of development occurs along with the elaboration of sleep-wake cycles, specifically a decline in REM sleep and an increase in NREM sleep, with circulatory control differing significantly between the two sleep phases. This chapter will describe the postnatal development of cardiovascular function and its physiological control in sleep during these critical early life phases.

STUDY OBJECTIVES: sleep deeply affects cardiac autonomic control, the impairment of which is associated with cardiovascular mortality. Obesity entails increased cardiovascular risk and derangements in sleep and cardiac autonomic control. We investigated whether cardiac autonomic control is impaired during sleep in ob/ob mice with morbid obesity caused by congenital leptin deficiency. DESIGN: indexes of cardiac autonomic control based on spontaneous cardiovascular fluctuations were compared between ob/ob and lean wild-type (+/+) mice during wakefulness, non-rapid eye movement sleep (NREMS), and rapid eye movement sleep (REMS). SETTING: N/A PATIENTS OR PARTICIPANTS: 7 ob/ob and 11 +/+ male mice. INTERVENTIONS: instrumentation with electrodes for sleep recordings and a telemetric transducer for measuring blood pressure and heart period. MEASUREMENTS AND RESULTS: In ob/ob mice, the variability of heart period and cardiac baroreflex sensitivity (sequence technique) were significantly lower than in +/+ mice during each wake-sleep state. The vagal modulation of heart period was significantly weaker in ob/ob than in +/+ mice during NREMS and REMS. In ob/ob mice, the cross-correlation function between heart period and blood pressure suggested that the baroreflex contribution to cardiac control was lower than in +/+ mice during wakefulness and NREMS, whereas the contribution of central autonomic commands was lower than in +/+ mice during NREMS and REMS. CONCLUSIONS: These data indicate a dysregulation of cardiac autonomic control during sleep in ob/ob mice. Ob/ob mice may represent a useful tool to understand the molecular pathways that lead to cardiac autonomic dysregulation during sleep in obesity.