Sleep-wake sensitive mechanisms of adenosine release in the basal forebrain of rodents : an in vitro study

Adenosine acting in the basal forebrain is a key mediator of sleep homeostasis. Extracellular adenosine concentrations increase during wakefulness, especially during prolonged wakefulness and lead to increased sleep pressure and subsequent rebound sleep. The release of endogenous adenosine during the sleep-wake cycle has mainly been studied in vivo with microdialysis techniques. The biochemical changes that accompany sleep-wake status may be preserved in vitro. We have therefore used adenosine-sensitive biosensors in slices of the basal forebrain (BFB) to study both depolarization-evoked adenosine release and the steady state adenosine tone in rats, mice and hamsters. Adenosine release was evoked by high K+, AMPA, NMDA and mGlu receptor agonists, but not by other transmitters associated with wakefulness such as orexin, histamine or neurotensin. Evoked and basal adenosine release in the BFB in vitro exhibited three key features: the magnitude of each varied systematically with the diurnal time at which the animal was sacrificed; sleep deprivation prior to sacrifice greatly increased both evoked adenosine release and the basal tone; and the enhancement of evoked adenosine release and basal tone resulting from sleep deprivation was reversed by the inducible nitric oxide synthase (iNOS) inhibitor, 1400 W. These data indicate that characteristics of adenosine release recorded in the BFB in vitro reflect those that have been linked in vivo to the homeostatic control of sleep. Our results provide methodologically independent support for a key role for induction of iNOS as a trigger for enhanced adenosine release following sleep deprivation and suggest that this induction may constitute a biochemical memory of this state

Sleep deprivation (SD) on focal brain ischemia in the rat : effects of different SD protocols

Sleep-wake disturbances are frequently observed in stroke patients and are associated with poorer functional outcome. Until now the effects of sleep on stroke evolution are unknown. The purpose of the present study was to evaluate the effects of three sleep deprivation (SD) protocols on brain damages after focal cerebral ischemia in a rat model. Permanent occlusion of distal branches of the middle cerebral artery was induced in adult rats. The animals were then subjected to 6h SD, 12h SD or sleep disturbances (SDis) in which 3 x 12h sleep deprivation were performed by gentle handling. Infarct size and brain swelling were assessed by Cresyl violet staining, and the number of damaged cells was measured by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining. Behavioral tests, namely tape removal and cylinder tests, were performed for assessing sensorimotor function. In the 6h SD protocol, no significant difference (P > 0.05) was found either in infarct size (42.5 ± 30.4 mm3 in sleep deprived animals vs. 44.5 ± 20.5 mm3 in controls, mean ± s.d.), in brain swelling (10.2 ± 3.8 % in sleep deprived animals vs. 11.3 ± 2.0 % in controls) or in number of TUNEL-positive cells (21.7 ± 2.0/mm2 in sleep deprived animals vs. 23.0 ± 1.1/mm2 in controls). In contrast, 12h sleep deprivation increased infarct size by 40 % (82.8 ± 10.9 mm3 in SD group vs. 59.2 ± 13.9 mm3 in control group, P = 0.008) and number of TUNEL-positive cells by 137 % (46.8 ± 15/mm in SD group vs. 19.7 ± 7.7/mm2 in control group, P = 0.003). There was no significant difference (P > 0.05) in brain swelling (12.9 ± 6.3 % in sleep deprived animals vs. 11.6 ± 6.0 % in controls). The SDis protocol also increased infarct size by 76 % (3 x 12h SD 58.8 ± 20.4 mm3 vs. no SD 33.8 ± 6.3 mm3, P = 0.017) and number of TUNEL-positive cells by 219 % (32.9 ± 13.2/mm2 vs. 10.3 ± 2.5/mm2, P = 0.008). Brain swelling did not show any difference between the two groups (24.5 ± 8.4 % in SD group vs. 16.7 ± 8.9 % in control group, p > 0.05). Both behavioral tests did not show any concluding results. In summary, we demonstrate that sleep deprivation aggravates brain damages in a rat model of stroke. Further experiments are needed to unveil the mechanisms underlying these effects

Sleep deprivation and brain energy metabolism : in vivo studies in rats and humans

Sleep deprivation leads to increased subsequent sleep length and depth and to deficits in cognitive performance in humans. In animals extreme sleep deprivation is eventually fatal. The cellular and molecular mechanisms causing the symptoms of sleep deprivation are unclear. This thesis was inspired by the hypothesis that during wakefulness brain energy stores would be depleted, and they would be replenished during sleep. The aim of this thesis was to elucidate the energy metabolic processes taking place in the brain during sleep deprivation. Endogenous brain energy metabolite levels were assessed in vivo in rats and in humans in four separate studies (Studies I-IV). In the first part (Study I) the effects of local energy depletion on brain energy metabolism and sleep were studied in rats with the use of in vivo microdialysis combined with high performance liquid chromatography. Energy depletion induced by 2,4-dinitrophenol infusion into the basal forebrain was comparable to the effects of sleep deprivation: both increased extracellular concentrations of adenosine, lactate, and pyruvate, and elevated subsequent sleep. This result supports the hypothesis of a connection between brain energy metabolism and sleep. The second part involved healthy human subjects (Studies II-IV). Study II aimed to assess the feasibility of applying proton magnetic resonance spectroscopy (1H MRS) to study brain lactate levels during cognitive stimulation. Cognitive stimulation induced an increase in lactate levels in the left inferior frontal gyrus, showing that metabolic imaging of neuronal activity related to cognition is possible with 1H MRS. Study III examined the effects of sleep deprivation and aging on the brain lactate response to cognitive stimulation. No physiologic, cognitive stimulation-induced lactate response appeared in the sleep-deprived and in the aging subjects, which can be interpreted as a sign of malfunctioning of brain energy metabolism. This malfunctioning may contribute to the functional impairment of the frontal cortex both during aging and sleep deprivation. Finally (Study IV), 1H MRS major metabolite levels in the occipital cortex were assessed during sleep deprivation and during photic stimulation. N-acetyl-aspartate (NAA/H2O) decreased during sleep deprivation, supporting the hypothesis of sleep deprivation-induced disturbance in brain energy metabolism. Choline containing compounds (Cho/H2O) decreased during sleep deprivation and recovered to alert levels during photic stimulation, pointing towards changes in membrane metabolism, and giving support to earlier observations of altered brain response to stimulation during sleep deprivation. Based on these findings, it can be concluded that sleep deprivation alters brain energy metabolism. However, the effects of sleep deprivation on brain energy metabolism may vary from one brain area to another. Although an effect of sleep deprivation might not in all cases be detectable in the non-stimulated baseline state, a challenge imposed by cognitive or photic stimulation can reveal significant changes. It can be hypothesized that brain energy metabolism during sleep deprivation is more vulnerable than in the alert state. Changes in brain energy metabolism may participate in the homeostatic regulation of sleep and contribute to the deficits in cognitive performance during sleep deprivation.Valvotus lisää korvausunen määrää ja johtaa ihmisillä kognitiivisen suorituskyvyn heikkenemiseen. Eläimillä on havaittu äärimmilleen pitkitetyn valvotuksen johtavan lopulta kuolemaan. Unen puutteen aiheuttamien oireiden taustalla olevat solu- ja molekyylitason mekanismit tunnetaan puutteellisesti. Väitöskirja Sleep deprivation and brain energy metabolism in vivo studies in rats and humans on saanut innoituksensa hypoteesista, jonka mukaan aivojen energiavarastot ehtyisivät valveen ja palautuisivat ennalleen unen aikana. Työssä tutkittiin aivojen energia-aineenvaihdunnan muutoksia valvotuksen aikana rotilla ja ihmisillä. Ensimmäisessä osatyössä tutkittiin aivojen paikallisen energiavajeen vaikutuksia rottien uneen ja aivojen energia-aineenvaihduntaan. Kokeellinen energiavaje etuaivojen pohjaosissa oli verrattavissa unen puutteen vaikutuksiin: molemmat aiheuttivat energia-aineenvaihduntatuotteiden (adenosiinin, laktaatin ja pyruvaatin) solunulkoisten pitoisuuksien kasvua sekä korvausunen lisääntymistä. Muissa osatöissä tutkittiin ihmisiä. Toisessa osatyössä todettiin protonispektroskopialla (1H MRS) kognitiivisen tehtävän suorittamisen nostavan terveiden aivojen laktaattipitoisuutta paikallisesti vasemmassa otsalohkossa (ns. laktaattivaste). Kolmannessa osatyössä todettiin, että tämä laktaattivaste ei tule esiin ikääntyvillä eikä valvotetuilla koehenkilöillä. Voidaan tulkita, että laktaattivasteen puuttuminen johtuu normaalin energia-aineenvaihdunnan häiriintymisestä. Pitkittyneen valveen sekä ikääntymisen aikana havaitut otsalohkon toiminnan häiriöt saattavat osin selittyä tämän havainnon pohjalta. Viimeisessä osatyössä todettiin näköaivokuoren N-asetyyliaspartaattipitoisuuden laskevan unen puutteen aikana, mikä tukee hypoteesia unen puutteen aikaisesta aivojen energia-aineenvaihdunnan häiriöstä. Myös koliiniyhdisteiden määrä näköaivokuorella laski unen puutteen aikana mutta palautui lähtötasolle näköärsytyksen myötä. Jälkimmäinen havainto viittaa solukalvojen aineenvaihdunnan muutoksiin unen puutteen aikana ja tukee aiempia havaintoja aivojen ärsytysvasteen muuttumisesta valvotetuilla koehenkilöillä. Yhteenvetona väitöskirjatyön tulosten perusteella voidaan päätellä, että valvominen muuttaa aivojen energia-aineenvaihduntaa. Unen puutteen vaikutus aivojen energia-aineenvaihduntaan voi kuitenkin vaihdella eri aivoalueiden välillä. Vaikka muutos ei aina tulekaan ilmi lepotilassa, se voi ilmetä kognitiivisen tehtävän suorittamisen tai näköärsytyksen aikana. Voidaan olettaa, että aivojen energia-aineenvaihdunta on unen puutteen aikana haavoittuvaisempi kuin virkeänä. Aivojen energia-aineenvaihdunnan muutokset saattavat osallistua unen säätelyyn sekä vaikuttaa kognitiivisen suorituskyvyn heikkenemiseen unen puutteen aikana

BEHAVIOURAL STATE CYCLING AND RELATED CHANGES IN THE CEREBRAL BLOOD FLOW IN THE OVINE FETUS NEAR TERM

The low voltage (LV)/rapid eye movement (REM) behavioural state the high voltage(HV)/non-rapid eye movement (NREM) behavioural state are each suggested to have a unique functional role in fetal neurodevelopment, therefore requiring the existence of both behavioural states in appropriate proportions for optimal maturation. The present study examined the behavioural state cycling pattern in the ovine fetus near term and characterized the related changes in cerebral blood flow velocity (CBFV), utilizing a 20-MHz piezoelectric Doppler crystal transducer. Our results demonstrated the HV/NREM epoch duration to be positively correlated with that of the prior LV/REM epoch, as well as with that of the subsequent LV/REM epoch, suggesting a possible homeostatic behavioural state control mechanism in the fetus. Changes in CBFV were consistent with those previously demonstrated, suggesting the piezoelectric Doppler crystal transducer may be used to continuously measure CBFV under resting conditions and provide a relative measure of cerebral blood flow changes

Is it time to turn our attention toward central mechanisms for post-exertional recovery strategies and performance?

• Central fatigue is accepted as a contributor to overall athletic performance, yet little research directly investigates post-exercise recovery strategies targeting the brain • Current post-exercise recovery strategies likely impact on the brain through a range of mechanisms, but improvements to these strategies is needed • Research is required to optimize post-exercise recovery with a focus on the brain Post-exercise recovery has largely focused on peripheral mechanisms of fatigue, but there is growing acceptance that fatigue is also contributed to through central mechanisms which demands that attention should be paid to optimizing recovery of the brain. In this narrative review we assemble evidence for the role that many currently utilized recovery strategies may have on the brain, as well as potential mechanisms for their action. The review provides discussion of how common nutritional strategies as well as physical modalities and methods to reduce mental fatigue are likely to interact with the brain, and offer an opportunity for subsequent improved performance. We aim to highlight the fact that many recovery strategies have been designed with the periphery in mind, and that refinement of current methods are likely to provide improvements in minimizing brain fatigue. Whilst we offer a number of recommendations, it is evident that there are many opportunities for improving the research, and practical guidelines in this area

Daily rhythm of cerebral blood flow velocity

BACKGROUND: CBFV (cerebral blood flow velocity) is lower in the morning than in the afternoon and evening. Two hypotheses have been proposed to explain the time of day changes in CBFV: 1) CBFV changes are due to sleep-associated processes or 2) time of day changes in CBFV are due to an endogenous circadian rhythm independent of sleep. The aim of this study was to examine CBFV over 30 hours of sustained wakefulness to determine whether CBFV exhibits fluctuations associated with time of day. METHODS: Eleven subjects underwent a modified constant routine protocol. CBFV from the middle cerebral artery was monitored by chronic recording of Transcranial Doppler (TCD) ultrasonography. Other variables included core body temperature (CBT), end-tidal carbon dioxide (EtCO2), blood pressure, and heart rate. Salivary dim light melatonin onset (DLMO) served as a measure of endogenous circadian phase position. RESULTS: A non-linear multiple regression, cosine fit analysis revealed that both the CBT and CBFV rhythm fit a 24 hour rhythm (R(2 )= 0.62 and R(2 )= 0.68, respectively). Circadian phase position of CBT occurred at 6:05 am while CBFV occurred at 12:02 pm, revealing a six hour, or 90 degree difference between these two rhythms (t = 4.9, df = 10, p < 0.01). Once aligned, the rhythm of CBFV closely tracked the rhythm of CBT as demonstrated by the substantial correlation between these two measures (r = 0.77, p < 0.01). CONCLUSION: In conclusion, time of day variations in CBFV have an approximately 24 hour rhythm under constant conditions, suggesting regulation by a circadian oscillator. The 90 degree-phase angle difference between the CBT and CBFV rhythms may help explain previous findings of lower CBFV values in the morning. The phase difference occurs at a time period during which cognitive performance decrements have been observed and when both cardiovascular and cerebrovascular events occur more frequently. The mechanisms underlying this phase angle difference require further exploration

Impact of Sleep and Circadian Disruption on Energy Balance and Diabetes: A Summary of Workshop Discussions

A workshop was held at the National Institute for Diabetes and Digestive and Kidney Diseases with a focus on the impact of sleep and circadian disruption on energy balance and diabetes. The workshop identified a number of key principles for research in this area and a number of specific opportunities. Studies in this area would be facilitated by active collaboration between investigators in sleep/circadian research and investigators in metabolism/diabetes. There is a need to translate the elegant findings from basic research into improving the metabolic health of the American public. There is also a need for investigators studying the impact of sleep/circadian disruption in humans to move beyond measurements of insulin and glucose and conduct more in-depth phenotyping. There is also a need for the assessments of sleep and circadian rhythms as well as assessments for sleep-disordered breathing to be incorporated into all ongoing cohort studies related to diabetes risk. Studies in humans need to complement the elegant short-term laboratory-based human studies of simulated short sleep and shift work etc. with studies in subjects in the general population with these disorders. It is conceivable that chronic adaptations occur, and if so, the mechanisms by which they occur needs to be identified and understood. Particular areas of opportunity that are ready for translation are studies to address whether CPAP treatment of patients with pre-diabetes and obstructive sleep apnea (OSA) prevents or delays the onset of diabetes and whether temporal restricted feeding has the same impact on obesity rates in humans as it does in mice

Neuroprotection, Photoperiod, and Sleep

After an acquired brain injury, responses that induce cell death are activated; however, neuroprotective mechanisms are also activated. The relation between these responses determines the destination of the damaged tissue. This relation presents variations throughout the day; numerous studies have shown that the onset of a stroke occurs preferably in the morning. In the rat, ischemia causes more damage when it is induced during the night. The damage caused by a traumatic brain injury (TBI), in the rat, varies depending on the time of day it is induced. Minor behavioral damage has been reported when the TBI occurs during the night, a period that coincides with the wakefulness of the rat. It also has been observed that sleep deprivation accelerates the recovery. Our group has documented that this is due, in part, to a difference in the degree of activation of cannabinergic, GABAergyc, and glutamatergic systems

Attention in the Brain Under Conditions of Sub-Optimal Alertness: Neurobiological Effects and Individual Differences

Sleep deprivation (SD) is a prevalent problem in modern society, and one that can have serious adverse consequences for health and safety. Critically, even short periods of SD can lead to relatively large decrements in attention, which may in turn cause an individual to neglect important environmental stimuli. In this thesis, I report the results of three experiments designed to investigate the neural bases of attentional declines under conditions of sleep loss and mental fatigue. In two experiments using arterial spin labeled fMRI, a technique that enables the quantification of absolute levels of cerebral blood flow (CBF), it was found that CBF patterns in the resting brain differed significantly based on arousal levels (Study #1) and prior cognitive workload (Study #2). These findings are a departure from prior neuroimaging studies, which have typically taken neural activity during non-task periods as static and inseparable baseline. In a test of sustained attention, performance declines were observed both following SD (Study #1) and when performing the task for an extended period of time while well-rested (Study #2). These decrements were primarily mediated by hypoactivation in a fronto-parietal attentional circuit. Furthermore, resting baseline levels of cerebral blood flow in the thalamus and prefrontal cortex before the start of the task were predictive of interindividual differences in subsequent performance decline (Study #2). In Study #3, an experiment using standard BOLD fMRI, it was found that performance declines in a test of selective attention following SD were accompanied by reduced functional connectivity between top-down control areas and regions of ventral visual cortex, as well as reductions in activation to targets in object-selective areas. Taken together, these results further our understanding of the neural basis of attention under conditions when this system is taxed beyond its normal limits