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      SNR optimized 31P functional MRS to detect mitochondrial and extracellular pH change during visual stimulation

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          Abstract

          Summary

          Energy metabolism of the human visual cortex was investigated by performing 31P functional MRS.

          Introduction

          The human brain is known to be the main glucose demanding organ of the human body and neuronal activity can increase this energy demand. In this study we investigate whether alterations in pH during activation of the brain can be observed with MRS, focusing on the mitochondrial inorganic phosphate (Pi) pool as potential marker of energy demand.

          Methods

          Six participants were scanned with 16 consecutive 31P‐MRSI scans, which were divided in 4 blocks of 8:36 minutes of either rest or visual stimulation. Since the signals from the mitochondrial compartments of Pi are low, multiple approaches to achieve high SNR 31P measurements were combined. This included: a close fitting 31P RF coil, a 7 T‐field strength, Ernst angle acquisitions and a stimulus with a large visual angle allowing large spectroscopy volumes containing activated tissue.

          Results

          The targeted resonance downfield of the main Pi peak could be distinguished, indicating the high SNR of the 31P spectra. The peak downfield of the main Pi peak is believed to be connected to mitochondrial performance. In addition, a BOLD effect in the PCr signal was observed as a signal increase of 2–3% during visual stimulation as compared to rest. When averaging data over multiple volunteers, a small subtle shift of about 0.1 ppm of the downfield Pi peak towards the main Pi peak could be observed in the first 4 minutes of visual stimulation, but no longer in the 4 to 8 minute scan window. Indications of a subtle shift during visual stimulation were found, but this effect remains small and should be further validated.

          Conclusion

          Overall, the downfield peak of Pi could be observed, revealing opportunities and considerations to measure specific acidity (pH) effects in the human visual cortex.

          Abstract

          Energy metabolism of the human visual cortex was investigated by performing 31P functional MRS, focused on mitochondrial inorganic phosphate (Pi mi/ex) as potential marker of energy demand. Since the concentration is low, multiple approaches to acquire high SNR 31P measurements were combined. Results show that the mitochondrial Pi resonance could be detected. During visual stimulation a small subtle shift was observed, especially in the first 4 minutes. Overall, this reveals opportunities and considerations to measure pH effects in the visual cortex.

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          Most cited references37

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          Sugar for the brain: the role of glucose in physiological and pathological brain function.

          The mammalian brain depends upon glucose as its main source of energy, and tight regulation of glucose metabolism is critical for brain physiology. Consistent with its critical role for physiological brain function, disruption of normal glucose metabolism as well as its interdependence with cell death pathways forms the pathophysiological basis for many brain disorders. Here, we review recent advances in understanding how glucose metabolism sustains basic brain physiology. We synthesize these findings to form a comprehensive picture of the cooperation required between different systems and cell types, and the specific breakdowns in this cooperation that lead to disease. Copyright © 2013 Elsevier Ltd. All rights reserved.
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            pH difference across the outer mitochondrial membrane measured with a green fluorescent protein mutant.

            In this study we have generated a EYFP targeted to the mitochondrial intermembrane space (MIMS-EYFP) to determine for the first time the pH within this compartment. The fragment encoding HAI-tagged EYFP was fused with the C-terminus of glycerol-phosphate dehydrogenase, an integral protein of the inner mitochondrial membrane. Human ECV304 cells transiently transfected with MIMS-EYFP showed the typical mitochondrial network, co-localized with MitoTracker Red. Following the calibration procedure, an estimation of the pH value in the intermembrane space was obtained. This value (6.88+/-0.09) was significantly lower than that determined in the cytosol after transfection with a cytosolic EYFP (7.59+/-0.01). Further, the pH of the mitochondrial matrix, determined with a EYFP targeted to this subcompartment, was 0.9 pH units higher than that in the intermembrane space. In conclusion, MIMS-EYFP represents a novel powerful tool to monitor pH changes in the mitochondrial intermembrane space of live cells.
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              Sustained neuronal activation raises oxidative metabolism to a new steady-state level: evidence from 1H NMR spectroscopy in the human visual cortex.

              To date, functional 1H NMR spectroscopy has been utilized to report the time courses of few metabolites, primarily lactate. Benefiting from the sensitivity offered by ultra-high magnetic field (7 T), the concentrations of 17 metabolites were measured in the human visual cortex during two paradigms of visual stimulation lasting 5.3 and 10.6 mins. Significant concentration changes of approximately 0.2 micromol/g were observed for several metabolites: lactate increased by 23%+/-5% (P<0.0005), glutamate increased by 3%+/-1% (P<0.01), whereas aspartate decreased by 15%+/-6% (P<0.05). Glucose concentration also manifested a tendency to decrease during activation periods. The lactate concentration reached the new steady-state level within the first minute of activation and came back to baseline only after the stimulus ended. The changes of the concentration of metabolites implied a rise in oxidative metabolism to a new steady-state level during activation and indicated that amino-acid homeostasis is affected by physiological stimulation, likely because of an increased flux through the malate-aspartate shuttle.
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                Author and article information

                Contributors
                a.d.hendriks-6@umcutrecht.nl
                d.w.j.klomp-2@umcutrecht.nl
                Journal
                NMR Biomed
                NMR Biomed
                10.1002/(ISSN)1099-1492
                NBM
                Nmr in Biomedicine
                John Wiley and Sons Inc. (Hoboken )
                0952-3480
                1099-1492
                22 July 2019
                November 2019
                : 32
                : 11 ( doiID: 10.1002/nbm.v32.11 )
                : e4137
                Affiliations
                [ 1 ] Department of Radiology University Medical Center Utrecht Utrecht the Netherlands
                Author notes
                [*] [* ] Correspondence

                Arjan Hendriks and Dennis Klomp, UMC Utrecht, Room Q.02.4.312, Heidelberglaan 100 (P.O. Box 85500), 3584 CX Utrecht, The Netherlands.

                Email: a.d.hendriks-6@ 123456umcutrecht.nl ; d.w.j.klomp-2@ 123456umcutrecht.nl

                Author information
                https://orcid.org/0000-0002-0363-2471
                https://orcid.org/0000-0003-1123-2352
                Article
                NBM4137 NBM-18-0171.R2
                10.1002/nbm.4137
                6900119
                31329342
                1ccd7d37-a861-400f-9f59-95674f040b06
                © 2019 The Authors NMR in Biomedicine Published by John Wiley & Sons Ltd

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

                History
                : 27 July 2018
                : 11 June 2019
                : 12 June 2019
                Page count
                Figures: 8, Tables: 0, Pages: 10, Words: 4978
                Funding
                Funded by: Dutch Research Council (NWO) , open-funder-registry 10.13039/501100003246;
                Award ID: 13339
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                November 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.2 mode:remove_FC converted:05.12.2019

                Radiology & Imaging
                mitochondrial pi,31p mrs,fmrs,ph,phosphorus spectroscopy,visual cortex
                Radiology & Imaging
                mitochondrial pi, 31p mrs, fmrs, ph, phosphorus spectroscopy, visual cortex

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