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Contribution of Extracellular Glutamine as An Anaplerotic Substrate to Neuronal Metabolism: A Re-Evaluation by Multinuclear NMR Spectroscopy in Primary Cultured Neurons

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Abstract

Multinuclear NMR spectroscopy is used to investigate the effect of glutamine on neuronal glucose metabolism. Primary neurons were incubated with [1-13C]glucose in the absence or presence of glutamine (2 mM) and/or NH4Cl (5 mM). After ammonia-treatment, the concentrations of high-energy phosphates decreased up to 84% of control, which was aggravated in glutamine-containing medium (up to 42% of control). These effects could not be attributed to changes in mitochondrial glucose oxidation. Withdrawal of glutamine decreased amino acid concentrations, e.g. of glutamate to 53%, but also considerably lessened the 13C enrichment in [4-13C]glutamate to 8.3% of control, and decreased the 13C-enrichment in acetyl-CoA entering the Krebs cycle (P<0.001). Thus, although glutamine is potent in replenishing neuronal glutamate stores, glutamate formation is mainly attributed to its de novo synthesis from glucose. Furthermore, mitochondrial glucose metabolism strongly depends on the supply of carbons from glutamine, indicating that exogenous glutamine is a well-suited substrate to replenish neuronal Krebs cycle intermediates.

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References

  1. R. P. Shank M. H. Aprison (1988) Glutamate as a neurotransmitter E. Kvamme (Eds) Glutamate and Glutamine in Mammals CRC Press Boca Raton, Florida 3–20

    Google Scholar 

  2. M. D. Norenberg A. Martinez-Hernandez (1979) ArticleTitleFine structural location of glutamine synthetase in astrocytes of rat brain Brain Res. 161 303–310 Occurrence Handle10.1016/0006-8993(79)90071-4 Occurrence Handle31966 Occurrence Handle1:STN:280:DyaE1M%2FotlKrsQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  3. J. C. Watkins R. H. Evans (1981) ArticleTitleExcitatory amino acid transmitters Ann. Rev. Pharmacol. Toxicol. 21 165–204 Occurrence Handle1:CAS:528:DyaL3MXitVarsbo%3D Occurrence Handle10.1146/annurev.pa.21.040181.001121

    Article  CAS  Google Scholar 

  4. F. Fonnum (1984) ArticleTitleGlutamate: A neurotransmitter in mammalian brain J. Neurochem. 42 1–11 Occurrence Handle6139418 Occurrence Handle1:CAS:528:DyaL2cXksFektQ%3D%3D

    PubMed  CAS  Google Scholar 

  5. T. Storck S. Shulte K. Hofmann W. Stoffel (1992) ArticleTitleStructure, expression, and functional analysis of Na-dependent glutamate/aspartate transporter from rat brain Proc. Natl. Acad. Sci. USA 89 10955–10959 Occurrence Handle1279699 Occurrence Handle1:CAS:528:DyaK3sXks1Gqsro%3D

    PubMed  CAS  Google Scholar 

  6. G. Pines N. C. Danbolt M. Bjoras (1992) ArticleTitleCloning and expression of a rat brain l-glutamate transporter Nature 360 464–467 Occurrence Handle10.1038/360464a0 Occurrence Handle1448170 Occurrence Handle1:CAS:528:DyaK3sXkvVKgu7w%3D

    Article  PubMed  CAS  Google Scholar 

  7. M. S. Patel (1974) ArticleTitleThe relative significance of CO2-fixing enzymes in the metabolism of rat brain J. Neurochem. 22 717–224 Occurrence Handle4152139 Occurrence Handle1:CAS:528:DyaE2cXltFOksb0%3D

    PubMed  CAS  Google Scholar 

  8. R. P. Shank G. S. Benneti S. O. Freytag G. L. Campbell (1985) ArticleTitlePyruvate carboxylase: An astrocyte-specific enzyme implicated in the replenishment of amino acid neurotransmitter pools Brain Res. 329 364–367 Occurrence Handle10.1016/0006-8993(85)90552-9 Occurrence Handle3884090 Occurrence Handle1:CAS:528:DyaL2MXhs1ChtLg%3D

    Article  PubMed  CAS  Google Scholar 

  9. G. A. Dienel L. Hertz (2001) ArticleTitleGlucose and lactate metabolism during brain activation J. Neurosci. Res. 66 824–838 Occurrence Handle11746408 Occurrence Handle1:CAS:528:DC%2BD3MXpt1SgtrY%3D

    PubMed  CAS  Google Scholar 

  10. A. Gjedde S. Marrett (2001) ArticleTitleGlycolysis in neurons, not astrocytes, delays oxidative metabolism of human visual cortex during sustained checkerboard stimulation in vivo J. Cereb. Blood Fow Metab. 21 1384–1392 Occurrence Handle1:CAS:528:DC%2BD38XjvV2isQ%3D%3D

    CAS  Google Scholar 

  11. L. Hertz L. Peng (1992) ArticleTitleEnergy metabolism at the cellular level of the CNS Can. J. Physiol. Pharmacol. 70 145–S157

    Google Scholar 

  12. H. F. Bradford H. K. Ward A. J. Thomas (1987) ArticleTitleGlutamine a major substrate for nerve endings J. Neurochem. 30 1453–1459

    Google Scholar 

  13. B. Hassel U. Sonnewald F. Fonnum (1995) ArticleTitleGlial-neuronal interactions as studied by cerebral metabolism of [2-13C]acetate and [1-13C]glucose: An ex vivo 13C NMR spectroscopic study J. Neurochem. 64 2773–2782 Occurrence Handle7760058 Occurrence Handle1:CAS:528:DyaK2MXlsl2lt7s%3D

    PubMed  CAS  Google Scholar 

  14. B. Hassel U. Sonnewald (1995) ArticleTitleGlial formation of pyruvate and lactate from TCA cycle intermediates: Implications for the inactivation of transmitter amino acids? J. Neurochem. 65 2227–2234 Occurrence Handle7595511 Occurrence Handle1:CAS:528:DyaK2MXovFGmtbY%3D

    PubMed  CAS  Google Scholar 

  15. C. Richter-Landesberg (1988) ArticleTitleNerve growth factor-inducible, large external (NILE) glycoprotein in developing rat cerebral cells in culture Cell Tissue Res. 252 181–190

    Google Scholar 

  16. A. Brand C. Richter-Landsberg D. Leibfritz (1997) ArticleTitleMetabolism of acetate in rat brain neurons, astrocytes and cocultures: Metabolic interactions between neurons and glia cells, monitored by NMR spectroscopy Cell. Mol. Biol. 43 645–657 Occurrence Handle9298588 Occurrence Handle1:CAS:528:DyaK2sXlvVejs7o%3D

    PubMed  CAS  Google Scholar 

  17. J. Goa (1953) ArticleTitleA micro biuret method for protein determination Scan. J. Clin. Lab. Invest. 5 218–222 Occurrence Handle1:CAS:528:DyaG2cXhtFKmsw%3D%3D Occurrence Handle10.3109/00365515309094189

    Article  CAS  Google Scholar 

  18. C. Zwingmann C. Richter-Landsberg A. Brand D. Leibfritz (2000) ArticleTitleNMR spectroscopic study on the metabolic fate of [3–(13)C]alanine in astrocytes, neurons, and cocultures: Implications for glia-neuron interactions in neurotransmitter metabolism Glia 32 286–303 Occurrence Handle10.1002/1098-1136(200012)32:3<286::AID-GLIA80>3.0.CO;2-P Occurrence Handle11102969 Occurrence Handle1:STN:280:DC%2BD3M%2FpslWisA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  19. C. Zwingmann D. Leibfritz (2003) ArticleTitleRegulation of glial metabolism studied by 13C-NMR NMR Biomed. 16 370–399 Occurrence Handle10.1002/nbm.850 Occurrence Handle14679501 Occurrence Handle1:CAS:528:DC%2BD2cXkslCksA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  20. J. C. Portais P. Voisin M. Merle P. Canioni (1996) ArticleTitleGlucose and glutamine metabolism in C6 glioma cells studied by carbon 13C NMR Biochimie. 78 155–164 Occurrence Handle10.1016/0300-9084(96)89500-9 Occurrence Handle8831946 Occurrence Handle1:CAS:528:DyaK28XkvF2ktLk%3D

    Article  PubMed  CAS  Google Scholar 

  21. M. Martin J. C. Portals P. Voisin N. Rousse P. Canioni M. Merle (1995) ArticleTitleComparative analysis of 13C-enriched metabolites released in the medium of cerebellar and cortical astrocytes incubated with [1-13C]glucose Eur. J. Biochem. 231 697–703 Occurrence Handle10.1111/j.1432-1033.1995.tb20750.x Occurrence Handle7649170 Occurrence Handle1:CAS:528:DyaK2MXns1Oisbc%3D

    Article  PubMed  CAS  Google Scholar 

  22. H. Chan C. Zwingmann M. Pannunzio R. F. Butterworth (2003) ArticleTitleEffects of ammonia on high affinity glutamate uptake and glutamate transporter EAAT3 expression in cultured rat cerebellar granule cells Neurochem. Int. 43 137–146 Occurrence Handle12620282 Occurrence Handle1:CAS:528:DC%2BD3sXhs1Kht7o%3D

    PubMed  CAS  Google Scholar 

  23. A. C. Yu J. Drejer L. Hertz A. Schousboe (1983) ArticleTitlePyruvate carboxylase activity in primary cultures of astrocytes and neurons J. Neurochem. 41 1484–1487 Occurrence Handle6619879 Occurrence Handle1:CAS:528:DyaL3sXlvFahurk%3D

    PubMed  CAS  Google Scholar 

  24. A. Gjedde S. Marrett M. Vafaee (2002) ArticleTitleOxidative and nonoxidative metabolism of excited neurons and astrocytes J. Cereb. Blood Flow Metab. 22 1–14 Occurrence Handle11807388 Occurrence Handle1:CAS:528:DC%2BD38XlsFWmsg%3D%3D

    PubMed  CAS  Google Scholar 

  25. D. Attwell S. B. Laughlin (2001) ArticleTitleAn energy budget for signaling in the grey matter of the brain J. Cereb. Blood Flow Metab. 21 1133–1145 Occurrence Handle11598490 Occurrence Handle1:CAS:528:DC%2BD3MXot1Ggsbc%3D

    PubMed  CAS  Google Scholar 

  26. C. Rae N. Hare W. A. Bubb S. R. McEwan A. Broer J. A. McQuillan V. J. Balcar A. D. Conigrave S. Broer (2003) ArticleTitleInhibition of glutamine transport depletes glutamate and GABA neurotransmitter pools: Further evidence for metabolic compartmentation J. Neurochem. 85 503–514 Occurrence Handle12675927 Occurrence Handle1:CAS:528:DC%2BD3sXjtlyls7Y%3D

    PubMed  CAS  Google Scholar 

  27. H. S. Waagepetersen U. Sonnewald O. M. Larsson A. Schousboe (2000) ArticleTitleA possible role of alanine for ammonia transfer between astrocytes and glutamatergic neurons J. Neurochem. 75 471–479 Occurrence Handle10.1046/j.1471-4159.2000.0750471.x Occurrence Handle10899921 Occurrence Handle1:CAS:528:DC%2BD3cXltFCltrw%3D

    Article  PubMed  CAS  Google Scholar 

  28. N. Chatauret C. Zwingmann C. Rose D. Leibfritz R. F. Butterworth (2003) ArticleTitleEffects of hypothermia on brain glucose metabolism in acute liver failure: A H/C-nuclear magnetic resonance study Gastroenterology 125 815–824 Occurrence Handle10.1016/S0016-5085(03)01054-0 Occurrence Handle12949727 Occurrence Handle1:CAS:528:DC%2BD3sXnvVSiu7w%3D

    Article  PubMed  CAS  Google Scholar 

  29. M. Swain R. F. Butterworth A. T. Blei (1992) ArticleTitleAmmonia and related amino acids in the pathogenesis of brain edema in acute ischemic liver failure in rats Hepatology 15 449–453 Occurrence Handle1544626 Occurrence Handle1:CAS:528:DyaK38XhslWnsL0%3D

    PubMed  CAS  Google Scholar 

  30. T. Holmin C. D. Agardh G. Alinder P. Herlin B. Hultberg (1983) ArticleTitleThe influence of total hepatectomy on cerebral energy state, ammonia-related amino acids of the brain and plasma amino acids in the rat Eur. J. Clin. Invest. 13 215–220 Occurrence Handle6135613 Occurrence Handle1:CAS:528:DyaL3sXktlKkt7s%3D

    PubMed  CAS  Google Scholar 

  31. A. M. Mans M. R. DeJoseph R. A. Hawkins (1994) ArticleTitleMetabolic abnormalities and grade of encephalopathy in acute hepatic failure J. Neurochem. 63 1829–1838 Occurrence Handle7931339 Occurrence Handle1:CAS:528:DyaK2cXmsFart7g%3D

    PubMed  CAS  Google Scholar 

  32. N. E. Deutz R. A. Chamuleau A. A. Graaf Particlede W. M. Bovee R. Beer Particlede (1988) ArticleTitleIn vivo 31P NMR spectroscopy of the rat cerebral cortex during acute hepatic encephalopathy NMR Biomed. 1 101–106 Occurrence Handle3275025 Occurrence Handle1:STN:280:DyaK3czltF2jsg%3D%3D

    PubMed  CAS  Google Scholar 

  33. S. P. Bessman A. N. Bessman (1955) ArticleTitleThe cerebral and peripheral uptake of ammonia in liver disease with a hypothesis for the mechanism of hepatic coma J. Clin. Invest. 34 622–628 Occurrence Handle14367516 Occurrence Handle1:CAS:528:DyaG1MXmvVClsA%3D%3D Occurrence Handle10.1172/JCI103111

    Article  PubMed  CAS  Google Scholar 

  34. H. F. Bradford S. P. Rose (1967) ArticleTitleIonic accumulation and membrane properties of enriched preparation of neurons and glia from mammalian cerebral cortex J. Neurochem. 14 373–375 Occurrence Handle6020464 Occurrence Handle1:CAS:528:DyaF2sXhtVWrsL4%3D

    PubMed  CAS  Google Scholar 

  35. J. H. Quastel (1970) ArticleTitleTransport processes at the brain cell membrane Neurosci. Res. 3 1–41 Occurrence Handle1:CAS:528:DyaE3cXkslCnsLc%3D

    CAS  Google Scholar 

  36. V. L. Rao C. R. Murthy R. F. Butterworth (1992) ArticleTitleGlutamatergic synaptic dysfunction in hyperammonemic syndromes Metab. Brain Dis. 7 1–20 Occurrence Handle1608363 Occurrence Handle1:CAS:528:DyaK3sXkslah

    PubMed  CAS  Google Scholar 

  37. H. Moser (1987) ArticleTitleElectrophysiological evidence for ammonium as a substitute for potassium in activating the sodium pump in a crayfish sensory neuron Can. J. Physiol. Pharmacol. 65 141–155 Occurrence Handle3567730 Occurrence Handle1:CAS:528:DyaL2sXhtlGlsLk%3D

    PubMed  CAS  Google Scholar 

  38. M. Dolinska B. Zablocka U. Sonnewald J. Albrecht (2004) ArticleTitleGlutamine uptake and expression of mRNA’s of glutamine transporting proteins in mouse cerebellar and cerebral cortical astrocytes and neurons Neurochem. Int. 44 75–81 Occurrence Handle10.1016/S0197-0186(03)00123-2 Occurrence Handle12971909 Occurrence Handle1:CAS:528:DC%2BD3sXntVGnu7k%3D

    Article  PubMed  CAS  Google Scholar 

  39. W. Walz S. Mukerji (1988) ArticleTitleLactate release from cultured astrocytes and neurons: A comparison Glia 1 366–370 Occurrence Handle10.1002/glia.440010603 Occurrence Handle2976396 Occurrence Handle1:STN:280:DyaL1M7lsVKitQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  40. A. T. Blei (1991) ArticleTitleCerebral edema and intracranial hypertension in acute liver failure: Distinct aspects of the same problem Hepatology 13 376–379 Occurrence Handle10.1016/0270-9139(91)92455-H Occurrence Handle1847353 Occurrence Handle1:STN:280:DyaK3M7jvFSksQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  41. A. T. Blei S. Olafsson G. Therrien R. F. Butterworth (1994) ArticleTitleAmmonia-induced brain edema and intracranial hypertension in rats after portacaval anastomosis Hepatology 19 1437–1444 Occurrence Handle10.1016/0270-9139(94)90240-2 Occurrence Handle8188174 Occurrence Handle1:STN:280:DyaK2c3ksFWjsw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  42. C. Zwingmann R. F. Butterworth (2005) ArticleTitleAn update on the role of brain glutamine synthesis and ist relation to cell-specific energy metabolism in the hyperammonemic brain: Further studies using NMR spectroscopy Neurochem. Int. 47 19–30 Occurrence Handle15916833 Occurrence Handle1:CAS:528:DC%2BD2MXlsl2ktL8%3D

    PubMed  CAS  Google Scholar 

  43. J. Cordoba F. Sanpedro J. Alonso A. Rovira (2002) ArticleTitle 1H magnetic resonance in the study of hepatic encephalopathy in humans Metab. Brain Dis. 17 415–429 Occurrence Handle12602517 Occurrence Handle1:CAS:528:DC%2BD3sXislGqsw%3D%3D

    PubMed  CAS  Google Scholar 

  44. A. Rovira J. Cordoba N. Raguer J. Alonso (2002) ArticleTitleMagnetic resonance imaging measurement of brain edema in patients with liver disease: Resolution after transplantation Curr. Opin. Neurol. 15 731–737 Occurrence Handle10.1097/00019052-200212000-00012 Occurrence Handle12447113

    Article  PubMed  Google Scholar 

  45. A. Michalak C. Rose J. Butterworth R. F. Butterworth (1996) ArticleTitleNeuroactive amino acids and glutamate (NMDA) receptors in frontal cortex of rats with experimental acute liver failure Hepatology 24 908–913 Occurrence Handle8855196 Occurrence Handle1:CAS:528:DyaK28XmvFOitrs%3D

    PubMed  CAS  Google Scholar 

  46. R. Debernardi P. J. Magistretti L. Pellerin (1999) ArticleTitleTrans-inhibition of glutamate transport prevents excitatory amino acid-induced glycolysis in astrocytes Brain Res. 850 39–46 Occurrence Handle10.1016/S0006-8993(99)02022-3 Occurrence Handle10629746 Occurrence Handle1:CAS:528:DC%2BD3cXjtVagtw%3D%3D

    Article  PubMed  CAS  Google Scholar 

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  1. Department of Organic Chemistry, University of Bremen, Leobener Strasse, 28334, Bremen, Germany

    Touraj Shokati, Claudia Zwingmann & Dieter Leibfritz

  2. Département de Médicine, Centre de Recherche, Hôpital Saint-Luc, CHUM, Université de Montreal, Montreal, Quebec, Canada

    Claudia Zwingmann

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Shokati, T., Zwingmann, C. & Leibfritz, D. Contribution of Extracellular Glutamine as An Anaplerotic Substrate to Neuronal Metabolism: A Re-Evaluation by Multinuclear NMR Spectroscopy in Primary Cultured Neurons. Neurochem Res 30, 1269–1281 (2005). https://doi.org/10.1007/s11064-005-8798-8

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