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Titolo:
Intracellular compartmentation of pyruvate in primary cultures of corticalneurons as detected by C-13 NMR spectroscopy with multiple C-13 labels
Autore:
Cruz, F; Villalba, M; Garcia-Espinosa, MA; Ballesteros, P; Bogonez, E; Satrustegui, J; Cerdan, S;
Indirizzi:
Univ Autonoma Madrid, CSIC, Inst Invest Biomed, E-28029 Madrid, Spain UnivAutonoma Madrid Madrid Spain E-28029 Biomed, E-28029 Madrid, Spain Univ Autonoma Madrid, Ctr Biol Mol Severo Ochoa, Dept Mol Biol, E-28029 Madrid, Spain Univ Autonoma Madrid Madrid Spain E-28029 ol Biol, E-28029 Madrid, Spain UNED, Dept Quim Organ & Biol, Madrid, Spain UNED Madrid SpainUNED, Dept Quim Organ & Biol, Madrid, Spain
Titolo Testata:
JOURNAL OF NEUROSCIENCE RESEARCH
fascicolo: 5, volume: 66, anno: 2001,
pagine: 771 - 781
SICI:
0360-4012(200112)66:5<771:ICOPIP>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
BLOOD-BRAIN-BARRIER; C6 GLIOMA-CELLS; GLUCOSE-METABOLISM; CEREBRAL METABOLISM; LACTATE; ASTROCYTES; RATS; GLIA; GLUTAMATE; TRANSPORT;
Keywords:
cortical neurons; glucose; pyruvate; malate; C-13 NMR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
54
Recensione:
Indirizzi per estratti:
Indirizzo: Cerdan, S Univ Autonoma Madrid, CSIC, Inst Invest Biomed, C Arturo Duperier 4, E-28029 Madrid, Spain Univ Autonoma Madrid C Arturo Duperier 4 MadridSpain E-28029 n
Citazione:
F. Cruz et al., "Intracellular compartmentation of pyruvate in primary cultures of corticalneurons as detected by C-13 NMR spectroscopy with multiple C-13 labels", J NEUROSC R, 66(5), 2001, pp. 771-781

Abstract

The intracellular compartmentation of pyruvate in primary cultures of cortical neurons was investigated by high resolution C-13 NMR using mixtures ofdifferent pyruvate precursors conveniently labeled with C-13 or unlabeled. Cells were incubated with 1-5 MM (1-C-13, 1,2-C-13(2) or U-C-13(6)) glucose only or with mixtures containing 1.5 mM (1-C-13 or U-C-13(6)) glucose, 0.25-2.5 mM (2-C-13 or 3-C-13) pyruvate and 1 mM malate. Extracts from cells and incubation media were analyzed by C-13 NMR to determine the relative contributions of the different precursors to the intracellular pyruvate pool. When (C-13) glucose was used as the sole substrate fractional C-13 enrichments and C-13 lsotopomer populations in lactate and glutamate carbons were compatible with a unique intracellular pool of pyruvate. When mixtures of (C-13) glucose, (C-13) pyruvate and malate were used, however, the fractional C-13 enrichments of the C2 and C3 carbons of lactate were higher than those of the C2 and C3 carbons of alanine and depicted a different C-13 isotopomer distribution. Moreover, neurons incubated with 1 mM (1,2-C-13(2)) glucose and 0.25-5 mM (3-C-13) pyruvate produced exclusively (3-C-13) lactate, revealing that extracellular pyruvate is the unique precursor of lactate under these conditions. These results reveal the presence of two different pools of intracellular pyruvate; one derived from extracellular pyruvate, used mainly for lactate and alanine production and one derived from glucose used primarily for oxidation. A red-ox switch using the cytosolic NAD(+)/NADHratio is proposed to modulate glycolytic flux, controlling which one of the two pyruvate pools is metabolized in the tricarboxylic acid cycle when substrates more oxidized or reduced than glucose are used. (C) 2001 Wiley-Liss, Inc.

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Documento generato il 02/04/20 alle ore 02:59:18