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

Magnetic Resonance Spectroscopy Investigations of Alzheimer Disease

Alzheimer disease is a progressively devastating neurodegenerative disease of the brain that impairs cognition and is ultimately fatal. Cholinesterase inhibitors are the current standard treatment for Alzheimer disease and they can alleviate some of the symptoms and thus improve quality of life. Cognitive measures aid in the diagnosis and monitoring of individuals with Alzheimer disease, but they do not directly measure disease pathophysiology. The purpose of this thesis is to investigate metabolic changes measured with proton magnetic resonance spectroscopy within the hippocampus and posterior cingulate, two brain regions known to be effected in Alzheimer disease, following cholinesterase inhibitor treatment. Such treatment is aimed at increasing the deficit of acetylcholine in Alzheimer disease. Secondly, to develop a 7 Tesla proton magnetic resonance spectroscopy data acquisition and metabolite quantification protocol to be used for future studies. In one study, proton magnetic resonance spectroscopy at 4 Tesla was used to measure the effects of four months of galantamine treatment (a cholinesterase inhibitor). An increase in the excitatory neurotransmitter glutamate was detected in the right hippocampus, and was associated with increased cognitive performance. In a second study, proton magnetic resonance spectroscopy at 3 Tesla was used to measure the effects of rivastigmine (a second cholinesterase inhibitor). The ratio of the neuronal marker N-acetylaspartate to creatine was decreased in the bilateral posterior cingulate cortex, which was associated with cognition. Finally, a quantitative proton magnetic resonance spectroscopy protocol at 7 Tesla was developed that incorporates subject-specific macromolecule removal. Absolute in vivo metabolite concentrations measured were in agreement with previous studies, and this protocol is ideal for applications in diseased conditions where macromolecule contributions may deviate from the norm

Association Among Neurophysiology, Cognition And Mobility In Older Adults

Age-related structural and molecular changes in older adults have been shown to significantly affect their cognitive and motor functions (Trollor and Valenzuela, 2001). Given the growing population of older adults, it is imperative to bridge the gaps in scientific understanding of cognition and motor capabilities in healthy older adults’ population. This study investigates five major neurochemicals and their potential correlations with gait, balance, executive function, and attention in healthy older adults. Additionally, we explore the interplay between cognition and motor performance in our participants. Furthermore, we hypothesize that the neurochemical values of interest may serve as predictive indicators for motor performance and cognition in healthy older adults

Advances in proton MR spectroscopy for quantifying pain associated metabolic changes in the human brain

In this work non-invasive in vivo detection of excitatory neurotransmitter glutamate andother cortical metabolites and their changes in the presence of acute and chronic pain wasperformed in the human brain with proton magnetic resonance spectroscopy (1H-MRS).This information can be used to better understand biochemical processes of cerebral painprocessing. Following introductory material, the first part of this thesis describes theimplemented method for post-processing of MR spectroscopic data to estimate absoluteconcentrations of the brain metabolites by considering the heterogeneous tissue compositionin the spectroscopic voxel. Phantom and in vivo brain studies demonstrated theadvantage of this method by reduced inter-individual variation of calculated metabolicconcentrations as well as enhanced quantitation accuracy. The second part of this workpresents the implemented method for the stimulus triggered data sampling permittingthe acquisition of in vivo 1H-MR spectra with a time resolution of few seconds. It wasshown that this method enables detection of changes of the neurotransmitter glutamateinduced by short acute pain stimuli. Considering these data, it was possible to characterisechanges of the glutamatergig neurotransmission associated with the sensation ofthe acute pain. The third part describes in vivo measurements on chronic pain patientsand healthy controls aiming to evaluate the changes of several brain metabolites in thedifferent cerebral pain processing regions associated with chronic pain. Patients revealeddecreased concentrations of the metabolic cell density markers and neurotransmitters indicatingthe degenerative processes as well as neurotransmitter dysfunctions, respectively.Results of this thesis indicate that pain induced metabolic changes in the human brainare traceable with the 1H-MRS by using experimental environment as it is used in clinicalroutine. This offers a broad spectrum of further applications aiming to explore thecerebral pain processing as well as to improve the specificity of the diagnostic assessmentof the chronic pain disease.Die vorliegende Arbeit beschreibt die Anwendung der Protonenmagnetresonanzspektroskopie(1H-MRS) zum nicht invasiven Nachweis von schmerzinduzierten Änderungen des erregenden Neurotransmitters Glutamat sowie anderer Metaboliten im menschlichen Gehirn. Diese Informationen könnten zu einem tieferen Verständnis der biochemischen Prozesse während der zerebralen Schmerzverarbeitung beitragen. Nach einer kurzen Einführung in die Problematik der Schmerzforschung sowie in die Grundlagen der MRSTechnikwird eine im Rahmen dieser Arbeit implementierte Methode zur Berechnung absoluter Metabolitenkonzentrationen unter Berücksichtigung der heterogenen Gewebezusammensetzung im spektroskopischen Volumen beschrieben. Der Vorteil dieses Verfahrens in Bezug auf die Verbesserung der Quantifizierungsgenauigkeit wird anhand von Ergebnissen spektroskopischer Messungen in einem Phantom sowie in Gehirnen gesunder Probanden belegt. Der zweite Teil befasst sich mit der Implementierung einer Technik zur reizgetriggerten Akquisition von MR Spektren, welche eine Abtastung verschiedener Stimulationszustände mit einer zeitlichen Auflösung von wenigen Sekunden zulässt und somit die Detektion dynamischer Änderungen von Metaboliten im Gehirn ermöglicht. Durch die Anwendung dieser Methode bei Messungen an gesunden Probanden konnten Änderungen im Glutamatstoffwechsel infolge einer Stimulation mit kurzen akuten Schmerzreizen nachgewiesen werden. Im dritten Teil der Arbeit wird schließlich eine an gesunden Probanden und Patienten mit chronischen Schmerzen durchgeführte Studievorgestellt, innerhalb derer die Auswirkungen der Schmerzchronifizierung auf den Metabolismus in schmerzverarbeitenden kortikalen Regionen untersucht wurden. Die Ergebnisse dieser Studie belegen die Hypothese, dass chronischer Schmerz mit Veränderungen imNeurotransmitterstoffwechsel sowie mit degenerativen Prozessen auf zellulärer Ebene einhergeht. Zusammenfassend lässt sich sagen, dass es mit der 1H-MRS möglich ist, schmerzinduzierte Änderungen der Metaboliten im menschlichen Gehirn unter Verwendung von klinischen Standartverfahren zu quantifizieren. Dies wiederum eröffnet ein breites Feld für weitere Untersuchungen, welche zur Erforschung der zerebralen Schmerzverarbeitung sowie zur Verbesserung der Spezifität diagnostischer Verfahren bei chronischen Schmerzen beitragen könnten

A longitudinal, multi-parametric functional MRI study to determine age-related changes in the rodent brain

As the population ages, the incidence of age-related neurological diseases and cognitive decline increases. To further understand disease-related changes in brain function it is advantageous to examine brain activity changes in healthy aging rodent models to permit mechanistic investigation. Here, we examine the suitability, in rodents, of using a novel, minimally invasive anaesthesia protocol in combination with a functional MRI protocol to assess alterations in neuronal activity due to physiological aging. 11 Wistar Han female rats were studied at 7, 9, 12, 15 and 18 months of age. Under an intravenous infusion of propofol, animals underwent functional magnetic resonance imaging (fMRI) and functional magnetic resonance spectroscopy (fMRS) with forepaw stimulation to quantify neurotransmitter activity, and resting cerebral blood flow (CBF) quantification using arterial spin labelling (ASL) to study changes in neurovascular coupling over time. Animals showed a significant decrease in size of the active region with age (P [less than] 0.05). fMRS results showed a significant decrease in glutamate change with stimulation (?Glu) with age (P < 0.05), and ?Glu became negative from 12 months onwards. Global CBF remained constant for the duration of the study. This study shows age related changes in the blood oxygen level dependent (BOLD) response in rodents that correlate with those seen in humans. The results also suggest that a reduction in synaptic glutamate turnover with age may underlie the reduction in the BOLD response, while CBF is preserved

Glutamatergic Metabolites and Gray Matter Losses in Schizophrenia: A Longitudinal Study Using In Vivo Proton Magnetic Resonance Spectroscopy

Approximately one in hundred people suffer from schizophrenia. Current medications partially improve the symptoms. There is no cure. Glutamate, an excitatory neurotransmitter, is a possible cause of the schizophrenia symptoms. Excessive glutamate release eventually leads to neurodegeneration. Longitudinal studies are necessary to observe the neurodegenerative process. Seventeen schizophrenia patients and 17 healthy volunteers underwent proton magnetic resonance spectroscopy (MRS) and imaging to measure neurochemical and structural changes in vivo. Metabolite levels were measured from a 1.5cm3 voxel in the anterior cingulate and thalamus using the stimulated echo acquisition mode sequence. Gray matter (GM) was assessed with voxel-based morphometry and ANALYZE. Total glutamatergic metabolite (tGL), N-acetylaspartate (NAA), and GM were significantly decreased in schizophrenia over 80 months. Reduced tGL and NAA levels were significantly correlated with GM changes. tGL loss was negatively correlated with social functioning. Significantly decreased tGL levels were possibly associated with GM loss in the spectroscopy voxel. Metabolite signal-to-noise ratio, but not quantification, was decreased as a function of MR system age. These findings demonstrate the feasibility of long-term MRS studies and implications for the pathophysiology of schizophrenia. tGL and GM losses were consistent with neurodegeneration but the effects of an early neurodevelopmental lesion or the effects of chronic medication cannot be ruled out. Structural and metabolite changes in these patients implicate glutamate as a possible target of medication in this disorder. The association between tGL loss and social functioning suggests it might be possible to arrest deterioration with pharmaceuticals that target glutamate

Energy failure following traumatic brain injury: Potential mechanisms and impact of normobaric hyperoxia

Cerebral ischaemia is a frequent finding in post mortem studies following traumatic brain injury (TBI), but clinical studies using 15oxygen positron emission tomography (15O PET) suggest that classical ischaemia is uncommon beyond the first 24 hours after injury. Evidence of metabolic failure in the absence of classical ischaemia may represent ongoing neuronal dysfunction and progressive neuronal loss. Any therapeutic intervention that mitigates such metabolic derangements before they result in irreversible neuronal injury may improve tissue fate and improve the functional outcome for patients. Energy failure was spatially defined, characterised, and mapped using 15O and 18Fluoromisinidazole ([18F] FMISO) positron emission tomography. This enabled differentiation of classical ischaemia, diffusion hypoxia, and established infarction, and provided data on the dominant local mechanism at any given time after TBI. My thesis also aimed to examine the utility of diffusion tensor imaging and whole-brain proton MR spectroscopy (WB 1H MRS) as imaging biomarkers to investigate normobaric hyperoxia as a therapeutic option following traumatic brain injury (TBI). Using ([18F] FMISO PET evidence of tissue hypoxia consistent with microvascular ischaemia was found across the injured brain. The impact of normobaric hyperoxia (NBH) was examined in a clinical TBI cohort using diffusion tensor imaging and WB 1H MRS. Some evidence of benefit was found within the perilesional brain, but further studies should examine the value of a longer period of exposure to NBH and whether this has implications for functional outcome.AAGBI, MRC, Wellcome trus

Physiological underpinnings of healthy brain ageing

Changes in cerebral perfusion or metabolism can occur as a result of healthy ageing, and in conditions of impaired ageing such as mild cognitive impairment (MCI) or Alzheimer’s disease (AD). Overarchingly, this thesis aimed to explore physiological magnetic resonance imaging (MRI) measures to study both cerebral perfusion and metabolism in the healthy ageing brain. Specifically, arterial spin labelling (ASL) and functional magnetic resonance spectroscopy (1H-fMRS) were employed in the elucidation of healthy ageing. Investigation of cerebral functionality is clinically important, enabling understanding of healthy ageing and disease pathology beyond that provided by structural measures. Given the necessity for tightly-regulated tissue perfusion in the delivery of oxygen to the brain, assessment of brain perfusion can enable elucidation of related brain health. Firstly, this thesis focused on changes in brain perfusion within a cross-sectional retrospective cohort of healthy subjects. This study aimed to assess the utility of univariate and multivariate pattern analysis (MVPA) techniques, and determine whether spatial coefficient of variation (sCoV) measures – which provide a method for inferring spatial heterogeneity of blood flow from single post-label delay (PLD) ASL data – are more significantly associated with age than standard perfusion metrics (ml/100g/min values). The impact of data processing steps on quantification of perfusion was initially assessed. Particularly, the influence of partial volume effect (PVE) correction and how this affected quantification of cerebral perfusion was of interest. The relationship between measures of cerebral perfusion – in regions of interest, vascular territories, and grey matter – and age were assessed, before grey matter (GM) spatial covariance patterns were identified, with MVPA hypothesised to elucidate more subtle age-related change than univariate, voxel-wise methodology. The executive control network (ECN) was the only network exhibiting a significant decline in perfusion with age, after controlling for relevant covariates. Interestingly, whilst the PCA approach resulted in a pattern of both positive and negative associations with age across cerebral GM, the surviving clusters in voxel-wise approaches were deemed spurious. Five-fold cross validation of PCA findings was used to assess whether the resultant spatial covariance patterns were able to predict subject age. This prediction was successful, with related r2 values of between 0.5316 and 0.7297 (p < 0.001 for all), however validation of these findings in an unseen dataset is required. The utility of the sCoV metric was also compared with standard tissue perfusion values, finding that sCoV may be more closely associated with ageing than ml/100g/min in certain regions. Particularly, a significant increase in whole GM sCoV with age was notable, given the absence of significant changes in perfusion with age in the same region. Additionally, a MVPA approach was used to establish the complex unknown relationship between cerebral perfusion and the Montreal Cognitive Assessment (MoCA), before graph visualisation was used to further understand the regional relatedness of the spatial covariance pattern. PCA resulted in a model which provided a moderate explanation of the aforementioned relationship, but this may be improved by inclusion of additional covariates in subsequent work, such as those pertaining to genetic status, such as apolipoprotein E (APOE). This study also replicates an FDG PET cognitive resilience signature in an ASL cohort for the first time, with a trend towards declining perfusion with age found (p = .08). Lastly, as ageing is associated with metabolic failure in the brain, which is often investigated using methodology which employs ionising radiation, the final study was motivated to investigate possible metabolic markers of brain ageing which can be measured using MRI. Metabolic-functional coupling can be studied using functional stimulation, and functional magnetic resonance spectroscopy (fMRS) is perfectly poised to elucidate certain metabolic behaviour. Given the close relationship between glucose (Glc) – the key fuel for cerebral functionality – and lactate (Lac) metabolism, an optimised long echo time (TE) semi-localized by adiabatic selective refocusing (semi-LASER) sequence (TE=144ms) with optimised J-modulation selection at 7T was employed to assess the effects of age on the dynamic behaviour of Lac, and determine its absolute concentrations throughout the time course, whilst a visual stimulation paradigm was viewed. Successful quantification of metabolite concentrations – including Lac, tCr and tNAA – was achieved in both the young and old cohorts, and their Lac peaks clearly visually identifiable throughout the time course. A significant increase in Lac concentration was observed between rest and stimulation, but not stimulation and recovery, in the young cohort. No significant Lac time course changes were identified in the full old cohort. This thesis concluded by summarising and contextualising the key findings herein, and discussion of possible directions for further associated research. The findings of this thesis broaden the field of knowledge around healthy ageing, and therefore may contribute to subsequent translation efforts for both clinical diagnostics and treatment approaches

Brain Changes in Long-Term Zen Meditators Using Proton Magnetic Resonance Spectroscopy and Diffusion Tensor Imaging: A Controlled Study

Introduction: This work aimed to determine whether 1H magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), diffusion-weighted imaging (DWI) and diffusion tensor imaging (DTI) are correlated with years of meditation and psychological variables in long-term Zen meditators compared to healthy non-meditator controls. Materials and Methods: Design. Controlled, cross-sectional study. Sample. Meditators were recruited from a Zen Buddhist monastery. The control group was recruited from hospital staff. Meditators were administered questionnaires on anxiety, depression, cognitive impairment and mindfulness. 1H-MRS (1.5 T) of the brain was carried out by exploring four areas: both thalami, both hippocampi, the posterior superior parietal lobule (PSPL) and posterior cingulate gyrus. Predefined areas of the brain were measured for diffusivity (ADC) and fractional anisotropy (FA) by MR-DTI. Results: Myo-inositol (mI) was increased in the posterior cingulate gyrus and Glutamate (Glu), N-acetyl-aspartate (NAA) and N-acetyl-aspartate/Creatine (NAA/Cr) was reduced in the left thalamus in meditators. We found a significant positive correlation between mI in the posterior cingulate and years of meditation (r = 0.518; p = .019). We also found significant negative correlations between Glu (r =20.452; p = .045), NAA (r =20.617; p = .003) and NAA/Cr (r =20.448; P = .047) in the left thalamus and years of meditation. Meditators showed a lower Apparent Diffusion Coefficient (ADC) in the left posterior parietal white matter than did controls, and the ADC was negatively correlated with years of meditation (r =20.4850, p = .0066). Conclusions: The results are consistent with the view that mI, Glu and NAA are the most important altered metabolites. This study provides evidence of subtle abnormalities in neuronal function in regions of the white matter in meditators