Expression of the human isoform of glutamate dehydrogenase, hGDH2, augments TCA cycle capacity and oxidative metabolism of glutamate during glucose deprivation in astrocytes

Jakob D. Nissen, Kasper Lykke, Jaroslaw Bryk, Malin H. Stridh, Ioannis Zaganas, Dorte M. Skytt, Arne Schousboe, Lasse K. Bak, Wolfgang Enard, Svante Pääbo, Helle S. Waagepetersen

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Abstract

A key enzyme in brain glutamate homeostasis is glutamate dehydrogenase (GDH) which links carbohydrate and amino acid metabolism mediating glutamate degradation to CO2 and expanding tricarboxylic acid (TCA) cycle capacity with intermediates, i.e. anaplerosis. Humans express two GDH isoforms, GDH1 and 2, whereas most other mammals express only GDH1. hGDH1 is widely expressed in human brain while hGDH2 is confined to astrocytes. The two isoforms display different enzymatic properties and the nature of these supports that hGDH2 expression in astrocytes potentially increases glutamate oxidation and supports the TCA cycle during energy-demanding processes such as high intensity glutamatergic signaling. However, little is known about how expression of hGDH2 affects the handling of glutamate and TCA cycle metabolism in astrocytes. Therefore, we cultured astrocytes from cerebral cortical tissue of hGDH2-expressing transgenic mice. We measured glutamate uptake and metabolism using [3H]glutamate, while the effect on metabolic pathways of glutamate and glucose was evaluated by use of 13C and 14C substrates and analysis by mass spectrometry and determination of radioactively labeled metabolites including CO2, respectively. We conclude that hGDH2 expression increases capacity for uptake and oxidative metabolism of glutamate, particularly during increased workload and aglycemia. Additionally, hGDH2 expression increased utilization of branched-chain amino acids (BCAA) during aglycemia and caused a general decrease in oxidative glucose metabolism. We speculate, that expression of hGDH2 allows astrocytes to spare glucose and utilize BCAAs during substrate shortages. These findings support the proposed role of hGDH2 in astrocytes as an important fail-safe during situations of intense glutamatergic activity.

LanguageEnglish
Pages474-488
Number of pages15
JournalGLIA
Volume65
Issue number3
Early online date29 Dec 2016
DOIs
Publication statusPublished - Mar 2017

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Glutamate Dehydrogenase
Citric Acid Cycle
Astrocytes
Glutamic Acid
Protein Isoforms
Glucose
Branched Chain Amino Acids
Brain
Metabolic Networks and Pathways
Workload
Transgenic Mice
Mammals
Mass Spectrometry
Homeostasis
Carbohydrates
Amino Acids
Enzymes

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Nissen, Jakob D. ; Lykke, Kasper ; Bryk, Jaroslaw ; Stridh, Malin H. ; Zaganas, Ioannis ; Skytt, Dorte M. ; Schousboe, Arne ; Bak, Lasse K. ; Enard, Wolfgang ; Pääbo, Svante ; Waagepetersen, Helle S. / Expression of the human isoform of glutamate dehydrogenase, hGDH2, augments TCA cycle capacity and oxidative metabolism of glutamate during glucose deprivation in astrocytes. In: GLIA. 2017 ; Vol. 65, No. 3. pp. 474-488.
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abstract = "A key enzyme in brain glutamate homeostasis is glutamate dehydrogenase (GDH) which links carbohydrate and amino acid metabolism mediating glutamate degradation to CO2 and expanding tricarboxylic acid (TCA) cycle capacity with intermediates, i.e. anaplerosis. Humans express two GDH isoforms, GDH1 and 2, whereas most other mammals express only GDH1. hGDH1 is widely expressed in human brain while hGDH2 is confined to astrocytes. The two isoforms display different enzymatic properties and the nature of these supports that hGDH2 expression in astrocytes potentially increases glutamate oxidation and supports the TCA cycle during energy-demanding processes such as high intensity glutamatergic signaling. However, little is known about how expression of hGDH2 affects the handling of glutamate and TCA cycle metabolism in astrocytes. Therefore, we cultured astrocytes from cerebral cortical tissue of hGDH2-expressing transgenic mice. We measured glutamate uptake and metabolism using [3H]glutamate, while the effect on metabolic pathways of glutamate and glucose was evaluated by use of 13C and 14C substrates and analysis by mass spectrometry and determination of radioactively labeled metabolites including CO2, respectively. We conclude that hGDH2 expression increases capacity for uptake and oxidative metabolism of glutamate, particularly during increased workload and aglycemia. Additionally, hGDH2 expression increased utilization of branched-chain amino acids (BCAA) during aglycemia and caused a general decrease in oxidative glucose metabolism. We speculate, that expression of hGDH2 allows astrocytes to spare glucose and utilize BCAAs during substrate shortages. These findings support the proposed role of hGDH2 in astrocytes as an important fail-safe during situations of intense glutamatergic activity.",
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Nissen, JD, Lykke, K, Bryk, J, Stridh, MH, Zaganas, I, Skytt, DM, Schousboe, A, Bak, LK, Enard, W, Pääbo, S & Waagepetersen, HS 2017, 'Expression of the human isoform of glutamate dehydrogenase, hGDH2, augments TCA cycle capacity and oxidative metabolism of glutamate during glucose deprivation in astrocytes' GLIA, vol. 65, no. 3, pp. 474-488. https://doi.org/10.1002/glia.23105

Expression of the human isoform of glutamate dehydrogenase, hGDH2, augments TCA cycle capacity and oxidative metabolism of glutamate during glucose deprivation in astrocytes. / Nissen, Jakob D.; Lykke, Kasper; Bryk, Jaroslaw; Stridh, Malin H.; Zaganas, Ioannis; Skytt, Dorte M.; Schousboe, Arne; Bak, Lasse K.; Enard, Wolfgang; Pääbo, Svante; Waagepetersen, Helle S.

In: GLIA, Vol. 65, No. 3, 03.2017, p. 474-488.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Expression of the human isoform of glutamate dehydrogenase, hGDH2, augments TCA cycle capacity and oxidative metabolism of glutamate during glucose deprivation in astrocytes

AU - Nissen,Jakob D.

AU - Lykke,Kasper

AU - Bryk,Jaroslaw

AU - Stridh,Malin H.

AU - Zaganas,Ioannis

AU - Skytt,Dorte M.

AU - Schousboe,Arne

AU - Bak,Lasse K.

AU - Enard,Wolfgang

AU - Pääbo,Svante

AU - Waagepetersen,Helle S.

N1 - Not OA compliant. No full text in Eprints HN 07/09/2017

PY - 2017/3

Y1 - 2017/3

N2 - A key enzyme in brain glutamate homeostasis is glutamate dehydrogenase (GDH) which links carbohydrate and amino acid metabolism mediating glutamate degradation to CO2 and expanding tricarboxylic acid (TCA) cycle capacity with intermediates, i.e. anaplerosis. Humans express two GDH isoforms, GDH1 and 2, whereas most other mammals express only GDH1. hGDH1 is widely expressed in human brain while hGDH2 is confined to astrocytes. The two isoforms display different enzymatic properties and the nature of these supports that hGDH2 expression in astrocytes potentially increases glutamate oxidation and supports the TCA cycle during energy-demanding processes such as high intensity glutamatergic signaling. However, little is known about how expression of hGDH2 affects the handling of glutamate and TCA cycle metabolism in astrocytes. Therefore, we cultured astrocytes from cerebral cortical tissue of hGDH2-expressing transgenic mice. We measured glutamate uptake and metabolism using [3H]glutamate, while the effect on metabolic pathways of glutamate and glucose was evaluated by use of 13C and 14C substrates and analysis by mass spectrometry and determination of radioactively labeled metabolites including CO2, respectively. We conclude that hGDH2 expression increases capacity for uptake and oxidative metabolism of glutamate, particularly during increased workload and aglycemia. Additionally, hGDH2 expression increased utilization of branched-chain amino acids (BCAA) during aglycemia and caused a general decrease in oxidative glucose metabolism. We speculate, that expression of hGDH2 allows astrocytes to spare glucose and utilize BCAAs during substrate shortages. These findings support the proposed role of hGDH2 in astrocytes as an important fail-safe during situations of intense glutamatergic activity.

AB - A key enzyme in brain glutamate homeostasis is glutamate dehydrogenase (GDH) which links carbohydrate and amino acid metabolism mediating glutamate degradation to CO2 and expanding tricarboxylic acid (TCA) cycle capacity with intermediates, i.e. anaplerosis. Humans express two GDH isoforms, GDH1 and 2, whereas most other mammals express only GDH1. hGDH1 is widely expressed in human brain while hGDH2 is confined to astrocytes. The two isoforms display different enzymatic properties and the nature of these supports that hGDH2 expression in astrocytes potentially increases glutamate oxidation and supports the TCA cycle during energy-demanding processes such as high intensity glutamatergic signaling. However, little is known about how expression of hGDH2 affects the handling of glutamate and TCA cycle metabolism in astrocytes. Therefore, we cultured astrocytes from cerebral cortical tissue of hGDH2-expressing transgenic mice. We measured glutamate uptake and metabolism using [3H]glutamate, while the effect on metabolic pathways of glutamate and glucose was evaluated by use of 13C and 14C substrates and analysis by mass spectrometry and determination of radioactively labeled metabolites including CO2, respectively. We conclude that hGDH2 expression increases capacity for uptake and oxidative metabolism of glutamate, particularly during increased workload and aglycemia. Additionally, hGDH2 expression increased utilization of branched-chain amino acids (BCAA) during aglycemia and caused a general decrease in oxidative glucose metabolism. We speculate, that expression of hGDH2 allows astrocytes to spare glucose and utilize BCAAs during substrate shortages. These findings support the proposed role of hGDH2 in astrocytes as an important fail-safe during situations of intense glutamatergic activity.

KW - Amino acid

KW - Anaplerosis

KW - Brain

KW - Hypoglycemic

KW - Isotopic labeling

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