Evaluation of the importance of transamination versus deamination in astrocytic metabolism of [U-13C] glutamate

Westergaard, Niels; Drejer, Joan; Schousboe, Arne; Sonnewald, Ursula

doi:10.1002/(sici)1098-1136(199606)17:2<160::aid-glia7>3.0.co;2-6

Cited by 103 publications

(43 citation statements)

References 37 publications

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“…The contents of amino acids in these cells are similar to previously published values for astrocytes cultured from neonatal brain [32][33][34] albeit variations between studies exist. However, the present results indicate that these astrocytes are comparable to astrocytes derived from brains of newborn mice.…”

Section: Discussionsupporting

confidence: 83%

Characterization of Primary and Secondary Cultures of Astrocytes Prepared from Mouse Cerebral Cortex

et al. 2010

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Astrocyte cultures were prepared from cerebral cortex of new-born and 7-day-old mice and additionally, the cultures from new-born animals were passaged as secondary cultures. The cultures were characterized by immunostaining for the astrocyte markers glutamine synthetase (GS), glial fibrillary acidic protein, and the glutamate transporters EAAT1 and EAAT2. The cultures prepared from 7-day-old animals were additionally characterized metabolically using (13)C-labeled glucose and glutamate as well as (15)N-labeled glutamate as substrates. All types of cultures exhibited pronounced immunostaining of the astrocyte marker proteins. The metabolic pattern of the cultures from 7-day-old animals of the labeled substrates was comparable to that seen previously in astrocyte cultures prepared from new-born mouse brain showing pronounced glycolytic and oxidative metabolism of glucose. Glutamate was metabolized both via the GS pathway and oxidatively via the tricarboxylic acid cycle as expected. Additionally, glutamate underwent pronounced transamination to aspartate and alanine and the intracellular pools of alanine and pyruvate exhibited compartmentation. Altogether the results show that cultures prepared from cerebral cortex of 7-day-old mice have metabolic and functional properties indistinguishable from those of classical astrocyte cultures prepared from neocortex of new-born animals. This provides flexibility with regard to preparation and use of these cultures for a variety of purposes.

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Section: Discussionsupporting

confidence: 83%

Characterization of Primary and Secondary Cultures of Astrocytes Prepared from Mouse Cerebral Cortex

et al. 2010

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“…Indeed, GDH, which is an anaplerotic enzyme, induces oxidative deamination of glutamate in astrocytes, leading to ␣-ketoglutarate, which can enter the TCA cycle and supply cells with energetic metabolites, such as lactate (Westergaard et al, 1996;Sonnewald et al, 1997). Moreover, in human epileptic tissue, Sherwin et al (1984) have shown that an increase in GDH expression correlates with an increase in consumption of glucose and neuronal activity.…”

Section: Discussionmentioning

confidence: 95%

Specific alteration in the expression of glial fibrillary acidic protein, glutamate dehydrogenase, and glutamine synthetase in rats with genetic absence epilepsy

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et al. 2000

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“…Both neurons and glial cells are capable of metabolising glucose via oxidative phosphorylation into CO 2 and water (Itoh et al 2003). It is also known that glial cells can utilise glucose for the generation of glutamate via transamination of the TCA cycle intermediate a-ketoglutarate Westergaard et al 1996). Glutamate is the main excitatory neurotransmitter in the mammalian CNS and is therefore strictly regulated to prevent neuronal over-excitability and excitotoxicity (Choi 1988;Greene and Greenamyre 1996;Michaelis 1998).…”

Section: Immunocytochemistry Of N1e-115 Cells Confirms Protein Expresmentioning

confidence: 93%

The potential of metabolomic analysis techniques for the characterisation of α1-adrenergic receptors in cultured N1E-115 mouse neuroblastoma cells

et al. 2015

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Several studies of neuropathic pain have linked abnormal adrenergic signalling to the development and maintenance of pain, although the mechanisms underlying this are not yet fully understood. Metabolomic analysis is a technique that can be used to give a snapshot of biochemical status, and can aid in the identification of the mechanisms behind pathological changes identified in cells, tissues and biological fluids. This study aimed to use gas chromatographymass spectrometry-based metabolomic profiling in combination with reverse transcriptase-polymerase chain reaction and immunocytochemistry to identify functional a 1 -adrenergic receptors on cultured N1E-115 mouse neuroblastoma cells. The study was able to confirm the presence of mRNA for the a 1D subtype, as well as protein expression of the a 1 -adrenergic receptor. Furthermore, metabolomic data revealed changes to the metabolite profile of cells when exposed to adrenergic pharmacological intervention. Agonist treatment with phenylephrine hydrochloride (10 lM) resulted in altered levels of several metabolites including myo-inositol, glucose, fructose, alanine, leucine, phenylalanine, valine, and n-acetylglutamic acid. Many of the changes observed in N1E-115 cells by agonist treatment were modulated by additional antagonist treatment (prazosin hydrochloride, 100 lM). A number of these changes reflected what is known about the biochemistry of a 1 -adrenergic receptor activation. This preliminary study therefore demonstrates the potential of metabolomic profiling to confirm the presence of functional receptors on cultured cells.

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Evaluation of the importance of transamination versus deamination in astrocytic metabolism of [U-13C] glutamate

Cited by 103 publications

References 37 publications

Characterization of Primary and Secondary Cultures of Astrocytes Prepared from Mouse Cerebral Cortex

Characterization of Primary and Secondary Cultures of Astrocytes Prepared from Mouse Cerebral Cortex

Specific alteration in the expression of glial fibrillary acidic protein, glutamate dehydrogenase, and glutamine synthetase in rats with genetic absence epilepsy

The potential of metabolomic analysis techniques for the characterisation of α1-adrenergic receptors in cultured N1E-115 mouse neuroblastoma cells

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