Glutamate receptor mediated cell death caused by glucose deprivation in NT2-N cells

Maureen M. O'Rourke, David E Pleasure

Research output: Contribution to journalArticle

Abstract

Introduction: We tested the hypothesis that NMDA or non-NMDA receptor mediated mechanisms may predominate to induce neuronal injury following glucose deprivation in human neuronal cells (NT2-N). The mechanisms of glucose deprivation induced neuronal cell death have been studied in numerous in vitro systems, most of which use fetal rat or mouse neurons not human neurons. Methods: NT2-N cells were cultured. Cells were washed with DMEM with 0 mM glucose, 25 mM HEPES, 4 mM glutamine, 100 IU/ml penicillin, 100 μg/ml streptomycin. Wells were divided into 4 groups: 1) Control-6 hours of glucose deprivation at 37°C and 0.21 FiO2 followed by normoglycemic recovery; 2) MK801 (NMDA receptor antagonist)-same as control except MK801 for 6 hours; 3) CNQX-(AMPA receptor antagonist)-same as control except CNQX for 6 hours; 4) MK801 + CNQX-same as control except MK801 and CNQX for 6 hours. Release of the cytoplasmic enzyme lactate dehydrogenase (LDH) was used as a measure of cell viability at 24 hours (LDH supernatant/LDH total × 100). Data were analyzed by ANOVA. Results: %LDH is less in the glucose-deprived neuronal cells exposed to MK801 (p<0.05), CNQX (p<0.05), and combined MK801 and CNQX (p<0.05) compared to control. (Figure Presented) Conclusions: Glucose deprivation causes a large release of LDH in NT2-N cells. NMDA receptor blockade as well as non-NMDA receptor blockade with CNQX decreases the release. Both NMDA and non-NMDA receptors play a mechanistic role in cell damage in hypoglycemia.

Original languageEnglish (US)
JournalCritical Care Medicine
Volume27
Issue number1 SUPPL.
StatePublished - 1999
Externally publishedYes

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6-Cyano-7-nitroquinoxaline-2,3-dione
Glutamate Receptors
Cell Death
L-Lactate Dehydrogenase
Glucose
N-Methylaspartate
N-Methyl-D-Aspartate Receptors
HEPES
Neurons
AMPA Receptors
Streptomycin
Glutamine
Hypoglycemia
Penicillins
Cultured Cells
Cell Survival
Analysis of Variance
Control Groups
Wounds and Injuries
Enzymes

ASJC Scopus subject areas

  • Critical Care and Intensive Care Medicine

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Glutamate receptor mediated cell death caused by glucose deprivation in NT2-N cells. / O'Rourke, Maureen M.; Pleasure, David E.

In: Critical Care Medicine, Vol. 27, No. 1 SUPPL., 1999.

Research output: Contribution to journalArticle

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abstract = "Introduction: We tested the hypothesis that NMDA or non-NMDA receptor mediated mechanisms may predominate to induce neuronal injury following glucose deprivation in human neuronal cells (NT2-N). The mechanisms of glucose deprivation induced neuronal cell death have been studied in numerous in vitro systems, most of which use fetal rat or mouse neurons not human neurons. Methods: NT2-N cells were cultured. Cells were washed with DMEM with 0 mM glucose, 25 mM HEPES, 4 mM glutamine, 100 IU/ml penicillin, 100 μg/ml streptomycin. Wells were divided into 4 groups: 1) Control-6 hours of glucose deprivation at 37°C and 0.21 FiO2 followed by normoglycemic recovery; 2) MK801 (NMDA receptor antagonist)-same as control except MK801 for 6 hours; 3) CNQX-(AMPA receptor antagonist)-same as control except CNQX for 6 hours; 4) MK801 + CNQX-same as control except MK801 and CNQX for 6 hours. Release of the cytoplasmic enzyme lactate dehydrogenase (LDH) was used as a measure of cell viability at 24 hours (LDH supernatant/LDH total × 100). Data were analyzed by ANOVA. Results: {\%}LDH is less in the glucose-deprived neuronal cells exposed to MK801 (p<0.05), CNQX (p<0.05), and combined MK801 and CNQX (p<0.05) compared to control. (Figure Presented) Conclusions: Glucose deprivation causes a large release of LDH in NT2-N cells. NMDA receptor blockade as well as non-NMDA receptor blockade with CNQX decreases the release. Both NMDA and non-NMDA receptors play a mechanistic role in cell damage in hypoglycemia.",
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N2 - Introduction: We tested the hypothesis that NMDA or non-NMDA receptor mediated mechanisms may predominate to induce neuronal injury following glucose deprivation in human neuronal cells (NT2-N). The mechanisms of glucose deprivation induced neuronal cell death have been studied in numerous in vitro systems, most of which use fetal rat or mouse neurons not human neurons. Methods: NT2-N cells were cultured. Cells were washed with DMEM with 0 mM glucose, 25 mM HEPES, 4 mM glutamine, 100 IU/ml penicillin, 100 μg/ml streptomycin. Wells were divided into 4 groups: 1) Control-6 hours of glucose deprivation at 37°C and 0.21 FiO2 followed by normoglycemic recovery; 2) MK801 (NMDA receptor antagonist)-same as control except MK801 for 6 hours; 3) CNQX-(AMPA receptor antagonist)-same as control except CNQX for 6 hours; 4) MK801 + CNQX-same as control except MK801 and CNQX for 6 hours. Release of the cytoplasmic enzyme lactate dehydrogenase (LDH) was used as a measure of cell viability at 24 hours (LDH supernatant/LDH total × 100). Data were analyzed by ANOVA. Results: %LDH is less in the glucose-deprived neuronal cells exposed to MK801 (p<0.05), CNQX (p<0.05), and combined MK801 and CNQX (p<0.05) compared to control. (Figure Presented) Conclusions: Glucose deprivation causes a large release of LDH in NT2-N cells. NMDA receptor blockade as well as non-NMDA receptor blockade with CNQX decreases the release. Both NMDA and non-NMDA receptors play a mechanistic role in cell damage in hypoglycemia.

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