Metabolism of lactate in cultured GABAergic neurons studied by 13C nuclear magnetic resonance spectroscopy

Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism
H S WaagepetersenA Schousboe

Abstract

Primary cultures of mouse cerebral cortical neurons (GABAergic) were incubated for 4 hours in media without glucose containing 1.0 mmol/L [U-13C]lactate in the absence or presence of 0.5 mmol/L glutamine. Redissolved, lyophilized cell extracts were analyzed by 13C nuclear magnetic resonance spectroscopy to investigate neuronal metabolism of lactate and by HPLC for determination of the total amounts of glutamate (Glu), gamma-aminobutyric acid (GABA), and aspartate (Asp). The 13C nuclear magnetic resonance spectra of cell extracts exhibited multiplets for Glu, GABA, and Asp, indicating pronounced recycling of labeled tricarboxylic acid cycle constituents. There was extensive incorporation of 13C label into amino acids in neurons incubated without glutamine, with the percent enrichments being approximately 60% for Glu and Asp, and 27% for GABA. When 0.5 mmol/L glutamine was added to the incubation medium, the enrichments for Asp, Glu, and GABA were 25%, 35%, and 25%, respectively. This strongly suggests that glutamine is readily converted to Glu and Asp but that conversion to GABA may be complex. The observation that enrichment in GABA was identical in the absence and presence of glutamine whereas cycling was decreased in the pres...Continue Reading

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Citations

Dec 23, 2003·Neuroscience Research·Masaaki TanakaYasuyoshi Watanabe
Sep 26, 2013·Trends in Endocrinology and Metabolism : TEM·Owen Chan, Robert Sherwin
Jun 25, 2003·Biochemical and Biophysical Research Communications·Masaaki TanakaYasuyoshi Watanabe
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Methods Mentioned

BETA
nuclear magnetic
NMR
nuclear magnetic resonance
deamination

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