We are analyzing https://link.springer.com/article/10.1007/s11064-006-9161-4.

Title:
Complex Glutamate Labeling from [U-13C]glucose or [U-13C]lactate in Co-cultures of Cerebellar Neurons and Astrocytes | Neurochemical Research
Description:
Glutamate metabolism was studied in co-cultures of mouse cerebellar neurons (predominantly glutamatergic) and astrocytes. One set of cultures was superfused (90 min) in the presence of either [U-13C]glucose (2.5 mM) and lactate (1 mM) or [U-13C]lactate (1 mM) and glucose (2.5 mM). Other sets of cultures were incubated in medium containing [U-13C]lactate (1 mM) and glucose (2.5 mM) for 4 h. Regardless of the experimental conditions cell extracts were analyzed using mass spectrometry and nuclear magnetic resonance spectroscopy. 13C labeling of glutamate was much higher than that of glutamine under all experimental conditions indicating that acetyl-CoA from both lactate and glucose was preferentially metabolized in the neurons. Aspartate labeling was similar to that of glutamate, especially when [U-13C]glucose was the substrate. Labeling of glutamate, aspartate and glutamine was lower in the cells incubated with [U-13C]lactate. The first part of the pyruvate recycling pathway, pyruvate formation, was detected in singlet and doublet labeling of alanine under all experimental conditions. However, full recycling, detectable in singlet labeling of glutamate in the C-4 position was only quantifiable in the superfused cells both from [U-13C]glucose and [U-13C]lactate. Lactate and alanine were mostly uniformly labeled and labeling of alanine was the same regardless of the labeled substrate present and higher than that of lactate when superfused in the presence of [U-13C]glucose. These results show that metabolism of pyruvate, the precursor for lactate, alanine and acetyl-CoA is highly compartmentalized.
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Keywords {🔍}

article, google, scholar, pubmed, cas, astrocytes, schousboe, metabolism, sonnewald, neurons, waagepetersen, lactate, pyruvate, glutamate, cerebellar, cultures, glucose, spectroscopy, neurosci, neurochem, labeling, cultured, res, cells, recycling, primary, content, research, ucglucose, uclactate, mouse, spectrometry, alanine, metabolic, rat, culture, privacy, cookies, bak, mass, glutamine, access, acid, hertz, analysis, information, publish, search, cocultures, glutamatergic,

Topics {✒️}

month download article/chapter liquid chromatography-mass spectrometry 13c-enriched metabolites released [u-13c]glutamate metabolism cerebellar neuronal-astrocytic cultures complex glutamate labeling full article pdf extensive gaba synthesis glutamate-glutamine cycle activity gas-liquid chromatography privacy choices/manage cookies related subjects gamma-aminobutyric acid cerebellar granule cells 1h-nmr spectroscopy energy metabolism 13c-nmr spectroscopy 13c nmr spectroscopy astrocytic pyruvate recycling amino acid content generous financial support pyruvate recycling pathway article bak nmr spectroscopic study 1h nmr study c6 glioma cells european economic area experimental conditions indicating n-2-hydroxyethyl-piperazine goodman eh jr bakken ij organic chemical applications folin phenol reagent fine structural localization expert technical assistance novo nordisk foundations olav kyrres gate [1–13c]glucose metabolism tissue culture manual conditions privacy policy isotopically labeled molecules maintain neurotransmitter homeostasis garcia-espinosa ma author information authors multiple 13c labels de gannes fm [u-13c5] glutamine glia–neuron interactions perfused rat liver accepting optional cookies

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         headline:Complex Glutamate Labeling from [U-13C]glucose or [U-13C]lactate in Co-cultures of Cerebellar Neurons and Astrocytes
         description:Glutamate metabolism was studied in co-cultures of mouse cerebellar neurons (predominantly glutamatergic) and astrocytes. One set of cultures was superfused (90 min) in the presence of either [U-13C]glucose (2.5 mM) and lactate (1 mM) or [U-13C]lactate (1 mM) and glucose (2.5 mM). Other sets of cultures were incubated in medium containing [U-13C]lactate (1 mM) and glucose (2.5 mM) for 4 h. Regardless of the experimental conditions cell extracts were analyzed using mass spectrometry and nuclear magnetic resonance spectroscopy. 13C labeling of glutamate was much higher than that of glutamine under all experimental conditions indicating that acetyl-CoA from both lactate and glucose was preferentially metabolized in the neurons. Aspartate labeling was similar to that of glutamate, especially when [U-13C]glucose was the substrate. Labeling of glutamate, aspartate and glutamine was lower in the cells incubated with [U-13C]lactate. The first part of the pyruvate recycling pathway, pyruvate formation, was detected in singlet and doublet labeling of alanine under all experimental conditions. However, full recycling, detectable in singlet labeling of glutamate in the C-4 position was only quantifiable in the superfused cells both from [U-13C]glucose and [U-13C]lactate. Lactate and alanine were mostly uniformly labeled and labeling of alanine was the same regardless of the labeled substrate present and higher than that of lactate when superfused in the presence of [U-13C]glucose. These results show that metabolism of pyruvate, the precursor for lactate, alanine and acetyl-CoA is highly compartmentalized.
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      description:Glutamate metabolism was studied in co-cultures of mouse cerebellar neurons (predominantly glutamatergic) and astrocytes. One set of cultures was superfused (90 min) in the presence of either [U-13C]glucose (2.5 mM) and lactate (1 mM) or [U-13C]lactate (1 mM) and glucose (2.5 mM). Other sets of cultures were incubated in medium containing [U-13C]lactate (1 mM) and glucose (2.5 mM) for 4 h. Regardless of the experimental conditions cell extracts were analyzed using mass spectrometry and nuclear magnetic resonance spectroscopy. 13C labeling of glutamate was much higher than that of glutamine under all experimental conditions indicating that acetyl-CoA from both lactate and glucose was preferentially metabolized in the neurons. Aspartate labeling was similar to that of glutamate, especially when [U-13C]glucose was the substrate. Labeling of glutamate, aspartate and glutamine was lower in the cells incubated with [U-13C]lactate. The first part of the pyruvate recycling pathway, pyruvate formation, was detected in singlet and doublet labeling of alanine under all experimental conditions. However, full recycling, detectable in singlet labeling of glutamate in the C-4 position was only quantifiable in the superfused cells both from [U-13C]glucose and [U-13C]lactate. Lactate and alanine were mostly uniformly labeled and labeling of alanine was the same regardless of the labeled substrate present and higher than that of lactate when superfused in the presence of [U-13C]glucose. These results show that metabolism of pyruvate, the precursor for lactate, alanine and acetyl-CoA is highly compartmentalized.
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         Mass spectrometry
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         Neuron
         Astrocyte
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         Neurochemistry
         Biochemistry
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         Cell Biology
         Neurology
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