bims-medebr Biomed News
on Metabolism of the developing brain
Issue of 2022–07–03
thirteen papers selected by
Regina F. Fernández, Johns Hopkins University



  1. Methods Mol Biol. 2022 ;2515 17-28
      Mitochondria are dynamic organelles that rely on a balance of opposing fission and fusion events to sustain mitochondrial function and efficiently meet the energy demands of a cell. As high-energy demanding cells, neurons rely heavily on optimally functional mitochondria with balanced mitochondrial dynamics, to ensure a sufficient energy supply required to maintain cell survival, establish membrane excitability and partake in processes of neurotransmission and plasticity. As such, many neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease) and stress conditions (e.g., stroke) leading to neuronal dysfunction or death are often associated with impaired mitochondrial function and dynamics, characterized by excessive mitochondrial fragmentation. For this reason, the assessment of mitochondrial morphology in neurons and within the brain can provide valuable information. The dynamic nature of mitochondria is not only observed in shape changes, but also changes in mitochondrial network connectivity and in cristae architecture. In this chapter, we will describe how mitochondrial morphology can be examined in vitro using hippocampal neuronal cultures and in vivo using mouse brain sections by immunocytochemistry, immunohistochemistry, and electron microscopy techniques.
    Keywords:  Confocal and electron microscopy; Cristae; Hippocampus; Mitochondrial dynamics; Mitochondrial dysfunction; Mitochondrial fission; Mitochondrial fusion; Mitochondrial morphology; Neurodegenerative diseases; Neuronal cultures
    DOI:  https://doi.org/10.1007/978-1-0716-2409-8_2
  2. Ann Agric Environ Med. 2022 Jun 24. pii: 145069. [Epub ahead of print]29(2): 201-206
       INTRODUCTION AND OBJECTIVE: The ketogenic diet (KD) is a high-fat, adequate-protein, low-carbohydrate diet that is getting more and more widespread in medicine. This dietary intervention causes changes in cerebral metabolism, which are considered potentially beneficial in patients with neurological disorders, but its impact should be controlled and assessed individually. The aim of this review is to provide an update of existing evidence concerning the utility of magnetic resonance spectroscopy (MRS) in monitoring shifts in the cerebral metabolism during ketogenic diet in patients with neurological disorders.
    REVIEW METHODS: The latest available literature was reviewed by May 13, 2021 using the PubMed, Web of Science and Google Scholar databases. There were 13 papers selected for analysis after reading the title, abstracts and whole text, meeting the assumed criteria.
    ABBREVIATED DESCRIPTION OF THE STATE OF KNOWLEDGE: MRS is a non-invasive imaging method providing information about the metabolism of brain tissues and playing an increasingly important role in monitoring the concentrations of cerebral metabolites in the course of such neurological disorders as primary brain tumour, epilepsy during KD. Recent trials prove that inverse correlation between serum β-hydroxybutyrate levels and N-acetylaspartate in brain tissue confirm antiepileptogenic properties of KD. Furthermore, ketone concentrations including β-hydroxybutyrate and acetone in both lesional and contralateral brain are referred to as correlating with average ketonuria in patients with primary brain tumou.
    SUMMARY: MRS is a feasible tool for detecting cerebral metabolic shifts linked to a ketogenic diet. However, further studies confirming MR spectroscopy utility in monitoring ketogenic diet treatment in patients with neurological disorders are needed.
    Keywords:  cerebral metabolism; ketogenic diet; magnetic resonance spectroscopy
    DOI:  https://doi.org/10.26444/aaem/145069
  3. Front Integr Neurosci. 2022 ;16 821850
      We report in a companion paper that in the mouse brain, in contrast to the 1,000-fold variation in local neuronal densities across sites, capillary density (measured both as capillary volume fraction and as density of endothelial cells) show very little variation, of the order of only fourfold. Here we confirm that finding in the rat brain and, using published rates of local blood flow and glucose use at rest, proceed to show that what small variation exists in capillary density across sites in the rat brain is strongly and linearly correlated to variations in local rates of brain metabolism at rest. Crucially, we show that such variations in local capillary density and brain metabolism are not correlated with local variations in neuronal density, which contradicts expectations that use-dependent self-organization would cause brain sites with more neurons to have higher capillary densities due to higher energetic demands. In fact, we show that the ratio of endothelial cells per neuron serves as a linear indicator of average blood flow and glucose use per neuron at rest, and both increase as neuronal density decreases across sites. In other words, because of the relatively tiny variation in capillary densities compared to the large variation in neuronal densities, the anatomical infrastructure of the brain is such that those sites with fewer neurons have more energy supplied per neuron, which matches a higher average rate of energy use per neuron, compared to sites with more neurons. Taken together, our data support the interpretation that resting brain metabolism is not demand-based, but rather limited by its capillary supply, and raise multiple implications for the differential vulnerability of diverse brain areas to disease and aging.
    Keywords:  brain energetics; brain vasculature; capillary density; metabolism; neuronal density
    DOI:  https://doi.org/10.3389/fnint.2022.821850
  4. J Neuropathol Exp Neurol. 2022 Jul 01. pii: nlac054. [Epub ahead of print]
      White matter degradation in the frontal lobe is one of the earliest detectable changes in aging and Alzheimer disease. The ε4 allele of apolipoprotein E (APOE4) is strongly associated with such myelin pathology but the underlying cellular mechanisms remain obscure. We hypothesized that, as a lipid transporter, APOE4 directly triggers pathology in the cholesterol-rich myelin sheath independent of AD pathology. To test this, we performed immunohistochemistry on brain tissues from healthy controls, sporadic, and familial Alzheimer disease subjects. While myelin basic protein expression was largely unchanged, in frontal cortex the number of oligodendrocytes (OLs) was significantly reduced in APOE4 brains independent of their Braak stage or NIA-RI criteria. This high vulnerability of OLs was confirmed in humanized APOE3 or APOE4 transgenic mice. A gradual decline of OL numbers was found in the aging brain without associated neuronal loss. Importantly, the application of lipidated human APOE4, but not APOE3, proteins significantly reduced the formation of myelinating OL in primary cell culture derived from Apoe-knockout mice, especially in cholesterol-depleted conditions. Our findings suggest that the disruption of myelination in APOE4 carriers may represent a direct OL pathology, rather than an indirect consequence of amyloid plaque formation or neuronal loss.
    Keywords:  APOE4; Alzheimer disease; Amyloid-independent; Lipid transport; Myelin; Oligodendrocyte
    DOI:  https://doi.org/10.1093/jnen/nlac054
  5. Front Aging Neurosci. 2022 ;14 893159
      Sporadic Alzheimer's disease (sAD) is the commonest cause of age-related neurodegeneration and dementia globally, and a leading cause of premature disability and death. To date, the quest for a disease-modifying therapy for sAD has failed, probably reflecting our incomplete understanding of aetiology and pathogenesis. Drugs that target aggregated Aβ/tau are ineffective, and metabolic defects are now considered to play substantive roles in sAD pathobiology. We tested the hypothesis that the recently identified, pervasive cerebral deficiency of pantothenate (vitamin B5) in sAD, might undermine brain energy metabolism by impairing levels of tricarboxylic acid (TCA)-cycle enzymes and enzyme complexes, some of which require the pantothenate-derived cofactor, coenzyme A (CoA) for their normal functioning. We applied proteomics to measure levels of the multi-subunit TCA-cycle enzymes and their cytoplasmic homologues. We analysed six functionally distinct brain regions from nine sAD cases and nine controls, measuring 33 cerebral proteins that comprise the nine enzymes of the mitochondrial-TCA cycle. Remarkably, we found widespread perturbations affecting only two multi-subunit enzymes and two enzyme complexes, whose function is modulated, directly or indirectly by CoA: pyruvate dehydrogenase complex, isocitrate dehydrogenase, 2-oxoglutarate dehydrogenase complex, and succinyl-CoA synthetase. The sAD cases we studied here displayed widespread deficiency of pantothenate, the obligatory precursor of CoA. Therefore, deficient cerebral pantothenate can damage brain-energy metabolism in sAD, at least in part through impairing levels of these four mitochondrial-TCA-cycle enzymes.
    Keywords:  coenzyme A (CoA); human brain; pantothenic acid/vitamin B5; pyruvate dehydrogenase complex; sporadic Alzheimer’s disease; tricarboxylic acid cycle (TCA cycle)
    DOI:  https://doi.org/10.3389/fnagi.2022.893159
  6. Methods Mol Biol. 2022 ;2497 63-72
      Mitochondria participate in many physiological and pathological processes in the cells, including cellular energy supply, regulation of calcium homeostasis, apoptosis, and ROS generation. Alterations of mitochondrial functions, especially the opening of mitochondrial permeability transition pore (mPTP) are the main mechanisms responsible for the ischemic brain damage. Recently, the inhibitors of the Complex I of mitochondrial respiratory chain emerged as promising suppressors of mitochondrial ROS generation and mPTP opening. Here we describe the assay that can be implemented easily to evaluate the protective effects of rotenone or other potential inhibitors of the Complex I of mitochondrial respiratory chain against acute ischemia-induced injuries in brain.
    Keywords:  Brain ischemia; Calcium retention capacity; Complex I; Complex II; Isolated brain mitochondria; Mitochondrial permeability transition pore; Mitochondrial respiration; ROS generation; Rotenone
    DOI:  https://doi.org/10.1007/978-1-0716-2309-1_3
  7. J Vis Exp. 2022 Jun 08.
      Mitochondria play an important role in cellular ATP production, reactive oxygen species regulation, and Ca2+ concentration control. Mitochondrial dysfunction has been implicated in the pathogenesis of multiple neurodegenerative diseases, including Parkinson's disease (PD), Huntington's disease, and Alzheimer's disease. To study the role of mitochondria in models of these diseases, we can measure mitochondrial respiration via oxygen consumption rate (OCR) as a proxy for mitochondrial function. OCR has already been successfully measured in cell cultures, as well as isolated mitochondria. However, these techniques are less physiologically relevant than measuring OCR in acute brain slices. To overcome this limitation, the authors developed a new method using a Seahorse XF analyzer to directly measure the OCR in acute striatal slices from adult mice. The technique is optimized with a focus on the striatum, a brain area involved in PD and Huntington's disease. The analyzer performs a live cell assay using a 24-well plate, which allows the simultaneous kinetic measurement of 24 samples. The method uses circular-punched pieces of striatal brain slices as samples. We demonstrate the effectiveness of this technique by identifying a lower basal OCR in striatal slices of a mouse model of PD. This method will be of broad interest to researchers working in the field of PD and Huntington's disease.
    DOI:  https://doi.org/10.3791/63379
  8. Neurotherapeutics. 2022 Jun 30.
      Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease that primarily affects motor neurons and causes muscle atrophy, paralysis, and death. While a great deal of progress has been made in deciphering the underlying pathogenic mechanisms, no effective treatments for the disease are currently available. This is mainly due to the high degree of complexity and heterogeneity that characterizes the disease. Over the last few decades of research, alterations to bioenergetic and metabolic homeostasis have emerged as a common denominator across many different forms of ALS. These alterations are found at the cellular level (e.g., mitochondrial dysfunction and impaired expression of monocarboxylate transporters) and at the systemic level (e.g., low BMI and hypermetabolism) and tend to be associated with survival or disease outcomes in patients. Furthermore, an increasing amount of preclinical evidence and some promising clinical evidence suggests that targeting energy metabolism could be an effective therapeutic strategy. This review examines the evidence both for and against these ALS-associated metabolic alterations and highlights potential avenues for therapeutic intervention.
    Keywords:  ALS; ATP; Glucose; Glycolysis; Lipids; Metabolism
    DOI:  https://doi.org/10.1007/s13311-022-01262-3
  9. Front Physiol. 2022 ;13 910567
      
    Keywords:  GPR81/HCAR1; MCT; brain; lactate; metabolism; muscle
    DOI:  https://doi.org/10.3389/fphys.2022.910567
  10. Compr Psychoneuroendocrinol. 2021 May;6 100055
       Aims: The communication between brain and peripheral homeostatic systems is a central element of ingestive control. We set out to explore which parts of the brain have strong functional connections to peripheral signalling molecules in a physiological context. It was hypothesised that associations can be found between endocrine response to glucose ingestion and preceding brain activity in dependence of the nutritional status of the body.
    Materials and methods: Young, healthy male participants underwent both a 38 ​h fasting and a control condition with standardized meals. On the second day of the experiment, participants underwent fMRI scanning followed by ingestion of glucose solution in both conditions. Subsequent endocrine responses relevant to energy metabolism were assessed. Associations between preceding brain activation and endocrine responses were examined.
    Results: In both fasting and non-fasting conditions, brain activity was associated with subsequent endocrine responses after glucose administration, but relevant brain areas differed substantially between the conditions. In the fasting condition relations between the caudate nucleus and the orbitofrontal regions with insulin and C-peptide were prevailing, whereas in the non-fasting condition associations between various brain regions and adiponectin and cortisol were the predominant significant outcome.
    Conclusion: Connections between endocrine response following a glucose challenge and prior brain activity suggests that the brain is playing an active role in the networks regulating food intake and associated endocrine signals. Further studies are needed to demonstrate causation.
    Keywords:  Cortisol; Food intake; Imaging; Magnetic resonance imaging
    DOI:  https://doi.org/10.1016/j.cpnec.2021.100055
  11. Nat Metab. 2022 Jun;4(6): 683-692
      Phospholipid levels are influenced by peripheral metabolism. Within the central nervous system, synaptic phospholipids regulate glutamatergic transmission and cortical excitability. Whether changes in peripheral metabolism affect brain lipid levels and cortical excitability remains unknown. Here, we show that levels of lysophosphatidic acid (LPA) species in the blood and cerebrospinal fluid are elevated after overnight fasting and lead to higher cortical excitability. LPA-related cortical excitability increases fasting-induced hyperphagia, and is decreased following inhibition of LPA synthesis. Mice expressing a human mutation (Prg-1R346T) leading to higher synaptic lipid-mediated cortical excitability display increased fasting-induced hyperphagia. Accordingly, human subjects with this mutation have higher body mass index and prevalence of type 2 diabetes. We further show that the effects of LPA following fasting are under the control of hypothalamic agouti-related peptide (AgRP) neurons. Depletion of AgRP-expressing cells in adult mice decreases fasting-induced elevation of circulating LPAs, as well as cortical excitability, while blunting hyperphagia. These findings reveal a direct influence of circulating LPAs under the control of hypothalamic AgRP neurons on cortical excitability, unmasking an alternative non-neuronal route by which the hypothalamus can exert a robust impact on the cortex and thereby affect food intake.
    DOI:  https://doi.org/10.1038/s42255-022-00589-7
  12. Glia. 2022 Jun 28.
      Oligodendrocytes (ODCs) are myelinating cells of the central nervous system (CNS) supporting neuronal survival. Oxidants and mitochondrial dysfunction have been suggested as the main causes of ODC damage during neuroinflammation as observed in multiple sclerosis (MS). Nonetheless, the dynamics of this process remain unclear, thus hindering the design of neuroprotective therapeutic strategies. To decipher the spatio-temporal pattern of oxidative damage and dysfunction of ODC mitochondria in vivo, we created a novel mouse model in which ODCs selectively express the ratiometric H2 O2 biosensor mito-roGFP2-Orp1 allowing for quantification of redox changes in their mitochondria. Using 2-photon imaging of the exposed spinal cord, we observed significant mitochondrial oxidation in ODCs upon induction of the MS model experimental autoimmune encephalomyelitis (EAE). This redox change became already apparent during the preclinical phase of EAE prior to CNS infiltration of inflammatory cells. Upon clinical EAE development, mitochondria oxidation remained detectable and was associated with a significant impairment in organelle density and morphology. These alterations correlated with the proximity of ODCs to inflammatory lesions containing activated microglia/macrophages. During the chronic progression of EAE, ODC mitochondria maintained an altered morphology, but their oxidant levels decreased to levels observed in healthy mice. Taken together, our study implicates oxidative stress in ODC mitochondria as a novel pre-clinical sign of MS-like inflammation and demonstrates that evolving redox and morphological changes in mitochondria accompany ODC dysfunction during neuroinflammation.
    Keywords:  experimental autoimmune encephalomyelitis; mitochondria; multiple sclerosis; myelin; neurodegeneration; oligodendrocyte; reactive species
    DOI:  https://doi.org/10.1002/glia.24235
  13. Methods Mol Biol. 2022 ;2497 73-81
      Mitochondrial impairment stands to be a major factor which contributes to the onset and pathogenesis of several neurodegenerative disorders, of which Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD) are among the notable ones. Extensive researches suggest the probable role of mitochondrial complex II and III dysfunction as underlying players in the pathogenesis of AD, PD, and HD. Present scenario of the world in occurrence of neurodegenerative disorders demands more research and development in this field. The development of enzyme histochemistry as an analytical technique has eased the assessment of mitochondrial complex activity at both qualitative and quantitative levels. Based on the principle of redox reactions of chromogenic substrates catalyzed by the enzymes in question, this histochemical analysis has been applied by researchers worldwide and has proved to be reliable. The present chapter hereby discusses the methods followed in performing histoenzymology of mitochondrial complex II and III activity. The chapter also puts light on the precautions which should be followed while performing histoenzymology in order to yield significant results.
    Keywords:  Alzheimer’s disease; Coenzyme Q-cytochrome c oxidoreductase; Enzyme histochemistry; Huntington’s disease; Mitochondrial complex dysfunction; Parkinson’s disease; Succinate dehydrogenase
    DOI:  https://doi.org/10.1007/978-1-0716-2309-1_4