Optimized fat oxidizing potentials -
Essentially, each compound was corrected in run-day blocks by registering the medians to equal one 1. Such evidence included 1 biological relevance given the genetic background context; 2 inclusion in a common pathway with a highly significant compound; 3 residing in a similar functional biochemical family with other significant compounds or 4 correlation with other experimental approaches.
Graphs corresponding to statistical analysis were carried out with GraphPad v. The resolved proteins were transferred electrophoretically to nitrocellulose membranes 0. The protein abundances of all western blots per condition were measured by densitometry of the bands, in the linear phase of the exposure without reaching saturation, on the films or on the scanned autoradiograms using ImageJ v.
At least three biologically independent replicates were always performed, although only one representative western blot is usually shown in the main figures. After electrophoresis, in-gel NADH dehydrogenase activity was evaluated allowing the identification of individual complex I and complex I-containing SC bands due to the formation of purple precipitated at the location of complex I ref.
Briefly, gels were incubated in 0. Next, a direct electrotransfer was performed followed by immunoblotting against mitochondrial complex I antibody NDUFS1 and complex III antibody UQCRC2. Mitochondrial ROS were determined with the fluorescent probe MitoSox Life Technologies.
MitoSox fluorescence intensity was assessed by flow cytometry FACScalibur flow cytometer, BD Biosciences and expressed in arbitrary units. For H 2 O 2 assessments, AmplexRed Life Technologies was used.
Slopes were used for calculations of the rates of H 2 O 2 formation. Brain blocks were rinsed three times with 0. After being rinsed with phosphate buffer solution, sections were mounted with Fluoromount Sigma aqueous mounting medium and cover slips Thermo Fisher.
Sections were examined with epifluorescence and the appropriate filter sets under an Operetta CLS high-content imaging system PerkinElmer.
An ANY-box core was used, which contained a light grey base and an adjustable perpendicular stick holding a camera and an infrared photo-beam array to track the animal movement and to detect rearing behaviour, respectively.
Mouse movements were tracked with the ANY-maze software and the AMi-maze interface to register all parameters described subsequently. A rotarod test Rotarod apparatus, Model , Ugo Basile was used to analyse motor balance and coordination. To score zone entries that consider the exploration of an object we consider the size of the object 3.
Along its perimeter there were 20 evenly spaced holes. The maze has one removable escape box that could be fitted under any of the holes and was filled with the animal bedding before each experiment. Black and white patterned pictures were used as spatial visual cues. The test consisted of three phases.
Finally, for the probe phase, mice were tested for spatial memory. In this session the escape box was removed, and the platform was virtually divided into four quadrants, each containing five holes. For other multiple-values comparisons, we used two-way analysis of variance ANOVA followed by Tukey tests.
All tests used are indicated in each figure legend. We carried out the statistical analysis using the GraphPad Prism v. The number of biologically independent culture preparations or animals used per experiment are indicated in the figure legends.
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.
Source data are provided with this paper. Hosli, L. et al. Direct vascular contact is a hallmark of cerebral astrocytes. Cell Rep. Article CAS PubMed Google Scholar.
Kacem, K. Structural organization of the perivascular astrocyte endfeet and their relationship with the endothelial glucose transporter: a confocal microscopy study. Glia 23 , 1—10 Almeida, A. Nitric oxide switches on glycolysis through the AMP protein kinase and 6-phosphofructokinase pathway.
Cell Biol. Barros, L. Why glucose transport in the brain matters for PET. Trends Neurosci. Magistretti, P. A cellular perspective on brain energy metabolism and functional imaging. Neuron 86 , — Zimmer, E. Article CAS PubMed PubMed Central Google Scholar. Bonvento, G. Astrocyte-neuron metabolic cooperation shapes brain activity.
Cell Metab. Eraso-Pichot, A. GSEA of mouse and human mitochondriomes reveals fatty acid oxidation in astrocytes. Glia 66 , — Article PubMed Google Scholar. Fecher, C. Cell-type-specific profiling of brain mitochondria reveals functional and molecular diversity.
Blazquez, C. Role of carnitine palmitoyltransferase I in the control of ketogenesis in primary cultures of rat astrocytes. Ioannou, M. Neuron-astrocyte metabolic coupling protects against activity-induced fatty acid toxicity. Cell , — e14 Qi, G. ApoE4 impairs neuron-astrocyte coupling of fatty acid metabolism.
Jogl, G. Crystal structure of carnitine acetyltransferase and implications for the catalytic mechanism and fatty acid transport. Cell , — Tong, L. Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery.
Life Sci. Barber, M. Structure and regulation of acetyl-CoA carboxylase genes of metazoa. Acta , 1—28 Wang, Y. Evidence for physical association of mitochondrial fatty acid oxidation and oxidative phosphorylation complexes.
Kruszynska, Y. Glucose kinetics during acute and chronic treatment of rats with 2[6 4-chloro-phenoxy hexyl]oxiranecarboxylate, etomoxir. Lopez-Fabuel, I.
Complex I assembly into supercomplexes determines differential mitochondrial ROS production in neurons and astrocytes. Natl Acad. USA , — Schoors, S. Fatty acid carbon is essential for dNTP synthesis in endothelial cells.
Nature , — Vicente-Gutierrez, C. An ex vivo approach to assess mitochondrial ROS by flow cytometry in AAV-tagged astrocytes in adult mice. Bio Protoc. Morillas, M. Structural model of the catalytic core of carnitine palmitoyltransferase I and carnitine octanoyltransferase COT : mutation of CPT I histidine and alanine and COT alanine impairs the catalytic activity.
Ding, L. Peroxisomal beta-oxidation acts as a sensor for intracellular fatty acids and regulates lipolysis. Fernandez-Vizarra, E. Two independent respiratory chains adapt OXPHOS performance to glycolytic switch. e6 Lapuente-Brun, E.
Supercomplex assembly determines electron flux in the mitochondrial electron transport chain. Science , — Lobo-Jarne, T. Respiratory chain supercomplexes: structures, function and biogenesis.
Cell Dev. Vercellino, I. The assembly, regulation and function of the mitochondrial respiratory chain. Maranzana, E. Mitochondrial respiratory supercomplex association limits production of reactive oxygen species from complex I.
Redox Signal 19 , — Astrocytic mitochondrial ROS modulate brain metabolism and mouse behaviour. The AMP-activated protein kinase is involved in the regulation of ketone body production by astrocytes.
Guzman, M. Is there an astrocyte-neuron ketone body shuttle? Trends Endocrinol. Schulz, J. Glial beta-oxidation regulates Drosophila energy metabolism. Silva, B. Glia fuel neurons with locally synthesized ketone bodies to sustain memory under starvation.
Pellerin, L. Glutamate uptake into astrocytes stimulates aerobic glycolysis: a mechanism coupling neuronal activity to glucose utilization. USA 91 , — Jimenez-Blasco, D. Glucose metabolism links astroglial mitochondria to cannabinoid effects. Bullitt, E. Expression of c-fos-like protein as a marker for neuronal activity following noxious stimulation in the rat.
Epstein, I. The Arc of cognition: signaling cascades regulating Arc and implications for cognitive function and disease. Carrasco, P. Ceramide levels regulated by carnitine palmitoyltransferase 1C control dendritic spine maturation and cognition.
The contribution of astrocytes to the 18Fdeoxyglucose signal in PET activation studies. Psychiatry 1 , — CAS PubMed Google Scholar. Hertz, L. Blood Flow. Dienel, G. Reevaluation of astrocyte-neuron energy metabolism with astrocyte volume fraction correction: impact on cellular glucose oxidation rates, glutamate-glutamine cycle energetics, glycogen levels and utilization rates vs.
Metabolic recruitment in brain tissue. Chan, K. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Yardeni, T. Retro-orbital injections in mice.
Article Google Scholar. Schonfeld, P. Brain energy metabolism spurns fatty acids as fuel due to their inherent mitotoxicity and potential capacity to unleash neurodegeneration.
Herrero-Mendez, A. Ragan, C. in Mitochondria: A Practical Approach eds Rickwood D. King, T. Preparation of succinate cytochrome c reductase and the cytochrome b-c1 particle, and reconstitution of succinate cytochrome c reductase. Methods Enzymol. Article CAS Google Scholar.
Wharton, D. Cytochrome oxidase from beef heart mitochondria. Shepherd, J. Exercise calorimetry in sedentary patients: procedures based on short 3 min steps underestimate carbohydrate oxidation and overestimate lipid oxidation. Diabetes Metab. PubMed Abstract CrossRef Full Text Google Scholar.
Brun, J. Sports 26, 57— Croci, I. Reproducibility of Fatmax and fat oxidation rates during exercise in recreationally trained males. PLoS One 9:e Frayn, K. Calculation of substrate oxidation rates in vivo from gaseous exchange.
Galgani, J. Metabolic flexibility and insulin resistance. Goodpaster, B. Metabolic flexibility in health and disease. Cell Metab. Hultman, E. Skeletal muscle energy metabolism and fatigue during intense exercise in man.
PubMed Abstract Google Scholar. Jeukendrup, A. Measurement of substrate oxidation during exercise by means of gas exchange measurements. Sports Med. MacFarlane, D. Open-circuit respirometry: a historical review of portable gas analysis systems.
Maunder, E. Contextualising maximal fat oxidation during exercise: determinants and normative values. Sanchez-Delgado, G. Activating brown adipose tissue through exercise ACTIBATE in young adults: rationale, design and methodology. Trials 45, — Shephard, R. Measurement of human energy expenditure, with particular reference to field studies: an historical perspective.
Stisen, A. Maximal fat oxidation rates in endurance trained and untrained women. Citation: Amaro-Gahete FJ, Sanchez-Delgado G, Helge JW and Ruiz JR Optimizing Maximal Fat Oxidation Assessment by a Treadmill-Based Graded Exercise Protocol: When Should the Test End? Received: 10 April ; Accepted: 02 July ; Published: 23 July Copyright © Amaro-Gahete, Sanchez-Delgado, Helge and Ruiz.
This is an open-access article distributed under the terms of the Creative Commons Attribution License CC BY. The use, distribution or reproduction in other forums is permitted, provided the original author s and the copyright owner s are credited and that the original publication in this journal is cited, in accordance with accepted academic practice.
No use, distribution or reproduction is permitted which does not comply with these terms. Amaro-Gahete, amarof ugr. Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers.
Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. Top bar navigation.
About us About us. Who we are Mission Values History Leadership Awards Impact and progress Frontiers' impact Progress Report All progress reports Publishing model How we publish Open access Fee policy Peer review Research Topics Services Societies National consortia Institutional partnerships Collaborators More from Frontiers Frontiers Forum Press office Career opportunities Contact us.
Sections Sections. About journal About journal. Article types Author guidelines Editor guidelines Publishing fees Submission checklist Contact editorial office. Endurance training induces a multitude of adaptations that result in increased fat oxidation.
The duration and intensity of exercise training required to induce changes in fat oxidation is currently unknown. Ingestion of carbohydrate in the hours before or on commencement of exercise reduces the rate of fat oxidation significantly compared with fasted conditions, whereas fasting longer than 6 h optimizes fat oxidation.
Fat oxidation rates have been shown to decrease after ingestion of high-fat diets, partly as a result of decreased glycogen stores and partly because of adaptations at the muscle level.
Whole-body fat oxidation pptentials exercise can be measured non-invasively during athlete profiling. Gaps in understanding exist in potetials relationships between fat oxidation during incremental fasted Body composition goals and skeletal O;timized parameters, potentialw performance, and fat Optimized fat oxidizing potentials during vat Optimized fat oxidizing potentials exercise. Seventeen Optkmized Optimized fat oxidizing potentials underwent a i fasted, incremental cycling test to assess peak whole-body fat oxidation PFOii resting vastus lateralis microbiopsy, and iii min maximal-effort cycling time-trial preceded by 2-h of fed-state moderate-intensity cycling to assess endurance performance and fed-state metabolism on separate occasions within one week. PFO 0. Addition of PFO to a traditional model of endurance peak oxygen uptake, power at 4 mmol. These associations suggest non-invasive measures of whole-body fat oxidation during exercise may be useful in the physiological profiling of endurance athletes.
Wacker, welche nötige Wörter..., der prächtige Gedanke
Nach meiner Meinung irren Sie sich. Schreiben Sie mir in PM.
Ja also!