Utilization of [15N]glutamate by Cultured Astrocytes

Overview

Journal Biochem J

Specialty Biochemistry

Date 1986 Feb 15

PMID 2871831

Citations 22

Authors

M Yudkoff

I Nissim

K Hummeler

M Medow

D Pleasure

Affiliations

Soon will be listed here.

Abstract

The metabolism of 0.25 mM-[15N]glutamic acid in cultured astrocytes was studied with gas chromatography-mass spectrometry. Almost all 15N was found as [2-15N]glutamine, [2-15N]glutamine, [5-15N]glutamine and [15N]alanine after 210 min of incubation. Some incorporation of 15N into aspartate and the 6-amino position of the adenine nucleotides also was observed, the latter reflecting activity of the purine nucleotide cycle. After the addition of [15N]glutamate the ammonia concentration in the medium declined, but the intracellular ATP concentration was unchanged despite concomitant ATP consumption in the glutamine synthetase reaction. Some potential sources of glutamate nitrogen were identified by incubating the astrocytes for 24 h with [5-15N]glutamine, [2-15N]glutamine or [15N]alanine. Significant labelling of glutamate was noted with addition of glutamine labelled on either the amino or the amide moiety, reflecting both glutaminase activity and reductive amination of 2-oxoglutarate in the glutamate dehydrogenase reaction. Alanine nitrogen also is an important source of glutamate nitrogen in this system.

Citing Articles

A metabolic profile of xenon and metabolite associations with 6-month mortality after out-of-hospital cardiac arrest: A post-hoc study of the randomised Xe-Hypotheca trial.

Nummela A, Scheinin H, Perola M, Joensuu A, Laitio R, Arola O PLoS One. 2024; 19(6):e0304966.

PMID: 38833442 PMC: 11149864. DOI: 10.1371/journal.pone.0304966.

Disturbance of the Glutamate-Glutamine Cycle, Secondary to Hepatic Damage, Compromises Memory Function.

Limon I, Angulo-Cruz I, Sanchez-Abdon L, Patricio-Martinez A Front Neurosci. 2021; 15:578922.

PMID: 33584185 PMC: 7873464. DOI: 10.3389/fnins.2021.578922.

Glutamine-Glutamate Cycle Flux Is Similar in Cultured Astrocytes and Brain and Both Glutamate Production and Oxidation Are Mainly Catalyzed by Aspartate Aminotransferase.

Hertz L, Rothman D Biology (Basel). 2017; 6(1).

PMID: 28245547 PMC: 5372010. DOI: 10.3390/biology6010017.

Glutamate pays its own way in astrocytes.

McKenna M Front Endocrinol (Lausanne). 2014; 4:191.

PMID: 24379804 PMC: 3863760. DOI: 10.3389/fendo.2013.00191.

De novo Synthesis of Glial Glutamate and Glutamine in Young Mice Requires Aspartate Provided by the Neuronal Mitochondrial Aspartate-Glutamate Carrier Aralar/AGC1.

Pardo B, Contreras L, Satrustegui J Front Endocrinol (Lausanne). 2013; 4:149.

PMID: 24133485 PMC: 3796713. DOI: 10.3389/fendo.2013.00149.

References

Shank R, Aprison M . Present status and significance of the glutamine cycle in neural tissues. Life Sci. 1981; 28(8):837-42. DOI: 10.1016/0024-3205(81)90044-8. View

Norenberg M . Distribution of glutamine synthetase in the rat central nervous system. J Histochem Cytochem. 1979; 27(3):756-62. DOI: 10.1177/27.3.39099. View

Freed B, Gelbard A . Distribution of 13N following intravenous injection of [13N]ammonia in the rat. Can J Physiol Pharmacol. 1982; 60(1):60-7. DOI: 10.1139/y82-008. View

RUSCAK M, Orlicky J, Zubor V, Hager H . Alanine aminotransferase in bovine brain: purification and properties. J Neurochem. 1982; 39(1):210-6. DOI: 10.1111/j.1471-4159.1982.tb04720.x. View

Ramaharobandro N, Borg J, Mandel P, Mark J . Glutamine and glutamate transport in cultured neuronal and glial cells. Brain Res. 1982; 244(1):113-21. DOI: 10.1016/0006-8993(82)90909-x. View

Drejer J, Larsson O, Schousboe A . Characterization of L-glutamate uptake into and release from astrocytes and neurons cultured from different brain regions. Exp Brain Res. 1982; 47(2):259-69. DOI: 10.1007/BF00239385. View

Yu A, Schousboe A, Hertz L . Metabolic fate of 14C-labeled glutamate in astrocytes in primary cultures. J Neurochem. 1982; 39(4):954-60. DOI: 10.1111/j.1471-4159.1982.tb11482.x. View

Gordon R, Balazs R . Characterization of separated cell types from the developing rat cerebellum: transport of glutamate and aspartate by preparations enriched in Purkinje cells, granule neurones, and astrocytes. J Neurochem. 1983; 40(4):1090-9. DOI: 10.1111/j.1471-4159.1983.tb08097.x. View

Yudkoff M, Nissim I, Kim S, Pleasure D, Hummeler K, Segal S . [15N] leucine as a source of [15N] glutamate in organotypic cerebellar explants. Biochem Biophys Res Commun. 1983; 115(1):174-9. DOI: 10.1016/0006-291x(83)90985-3. View

10.

Kim S, Stern J, Kim M, Pleasure D . Culture of purified rat astrocytes in serum-free medium supplemented with mitogen. Brain Res. 1983; 274(1):79-86. DOI: 10.1016/0006-8993(83)90522-x. View

11.

Jones B, Gilligan J . o-Phthaldialdehyde precolumn derivatization and reversed-phase high-performance liquid chromatography of polypeptide hydrolysates and physiological fluids. J Chromatogr. 1983; 266:471-82. DOI: 10.1016/s0021-9673(01)90918-5. View

12.

Yudkoff M, Nissim I, Kim S, Pleasure D, Segal S . Metabolism of 15NH3 in organotypic cerebellar explants and cultured astrocytes: studies with gas chromatography-mass spectrometry. J Neurochem. 1984; 42(1):283-6. DOI: 10.1111/j.1471-4159.1984.tb09731.x. View

13.

Yu A, Schousboe A, Hertz L . Influence of pathological concentrations of ammonia on metabolic fate of 14C-labeled glutamate in astrocytes in primary cultures. J Neurochem. 1984; 42(2):594-7. DOI: 10.1111/j.1471-4159.1984.tb02721.x. View

14.

Swain J, HINES J, Sabina R, Harbury O, Holmes E . Disruption of the purine nucleotide cycle by inhibition of adenylosuccinate lyase produces skeletal muscle dysfunction. J Clin Invest. 1984; 74(4):1422-7. PMC: 425310. DOI: 10.1172/JCI111553. View

15.

Nissim I, Yudkoff M, Lapidot A . Simultaneous determination of [2-15N]- and [5-15N]glutamine with gas chromatography-mass spectroscopy: applications to nitrogen metabolic studies. Anal Biochem. 1984; 143(1):14-20. DOI: 10.1016/0003-2697(84)90550-5. View

16.

Lewis S, Yudkoff M . Gas chromatographic-mass spectrometric determination of isotopic enrichment of 6-15NH2 in adenine nucleotides. Anal Biochem. 1985; 145(2):354-61. DOI: 10.1016/0003-2697(85)90374-4. View

17.

BERL S, TAKAGAKI G, Clarke D, WAELSCH H . Metabolic compartments in vivo. Ammonia and glutamic acid metabolism in brain and liver. J Biol Chem. 1962; 237:2562-9. View

18.

BERL S . Glutamine synthetase. Determination of its distribution in brain during development. Biochemistry. 1966; 5(3):916-22. DOI: 10.1021/bi00867a016. View

19.

Logan W, Snyder S . High affinity uptake systems for glycine, glutamic and aspaspartic acids in synaptosomes of rat central nervous tissues. Brain Res. 1972; 42(2):413-31. DOI: 10.1016/0006-8993(72)90540-9. View

20.

Henn F, GOLDSTEIN M, Hamberger A . Uptake of the neurotransmitter candidate glutamate by glia. Nature. 1974; 249(458):663-4. DOI: 10.1038/249663a0. View