Pathways involved in the generation of reactive oxygen and nitrogen species during glucose deprivation and its role on the death of cultured hippocampal neurons

Paramo, Blanca; Hernández-Fonseca, Karla; Estrada‐Sánchez, Ana María; Jiménez, N.; Hernández‐Cruz, Arturo; Massieu, Lourdes

doi:10.1016/j.neuroscience.2010.02.074

Cited by 42 publications

(33 citation statements)

References 47 publications

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“…Cell death induced by glucose deprivation was previously reported in SN56 cells [21,22] and neurons in culture [23]. SN56 is a line of cholinergic neurons which are highly sensitive to several stressors and to metabolic alterations.…”

Section: Measurements In Glucose Deprivationmentioning

confidence: 82%

Targeting of multiple metabolites in neural cells monitored by using protein-based carbon nanotubes

Boero

Carrara

Vecchio

et al. 2011

Sensors and Actuators B: Chemical

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a b s t r a c tMicrodevices dedicated to monitor metabolite levels have recently enabled many applications in the field of cell analysis, to monitor cell growth and development of numerous cell lines. By combining the traditional technology used for electrochemical biosensors with nanoscale materials, it is possible to develop miniaturized metabolite biosensors with unique properties of sensitivity and detection limit. In particular, enzymes tend to adsorb onto carbon nanotubes and their optical or electrical activity can perturb the electronic properties. In the present work we propose multi-walled carbon nanotube-based biosensors to monitor a cell line highly sensitive to metabolic alterations, in order to evaluate lactate production and glucose uptake during different cell states. We achieve sensors for both lactate and glucose, with sensitivities of 40.1 A mM −1 cm −2 and 27.7 A mM −1 cm −2 , and detection limits of 28 M and 73 M, respectively. This nano-biosensing technology is used to provide new information on cell line metabolism during proliferation and differentiation, which are unprecedented in cell biology.

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Section: Measurements In Glucose Deprivationmentioning

confidence: 82%

Targeting of multiple metabolites in neural cells monitored by using protein-based carbon nanotubes

Boero

Carrara

Vecchio

et al. 2011

Sensors and Actuators B: Chemical

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“…Cell survival was monitored after 18 hours recovery by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (Sigma-Aldrich) reduction assay and the lactic acid dehydrogenase release assay, as previously described. 20 Briefly, after 18 hours of GR cells were incubated with MTT (150 μmol/L) for 1 hour at 37°C in 5%CO 2 /95% air atmosphere; the medium was aspirated and the precipitated formazan salts were solubilized in acidic isopropanol. Absorbance was monitored spectrophotometrically at a wavelength of 570 nm.…”

Section: Cell Treatmentsmentioning

confidence: 99%

“…ATP concentration was determined immediately after 1.5 hours GD or after 3 hours GR. ATP levels were determined by means of a luminometer through the luceferinluciferase quimioluminescent kit (Molecular Probes, Eugene, OR, USA), as previously described 20 and ATP concentrations were calculated from readings obtained from an ATP standard curve (from 6.5 to 250 pmol). Aliquots of cell homogenates were kept for protein determination by the Lowry's method 26 and data are expressed as pmol/μg of protein.…”

Section: Atp Determinationmentioning

confidence: 99%

Protection of Hypoglycemia-Induced Neuronal Death by β-Hydroxybutyrate Involves the Preservation of Energy Levels and Decreased Production of Reactive Oxygen Species

Julio-Amilpas

Montiel

Soto‐Tinoco

et al. 2015

J Cereb Blood Flow Metab

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Glucose is the main energy substrate in brain but in certain circumstances such as prolonged fasting and the suckling period alternative substrates can be used such as the ketone bodies (KB), beta-hydroxybutyrate (BHB), and acetoacetate. It has been shown that KB prevent neuronal death induced during energy limiting conditions and excitotoxicity. The protective effect of KB has been mainly attributed to the improvement of mitochondrial function. In the present study, we have investigated the protective effect of D-BHB against neuronal death induced by severe noncoma hypoglycemia in the rat in vivo and by glucose deprivation (GD) in cortical cultures. Results show that systemic administration of D-BHB reduces reactive oxygen species (ROS) production in distinct cortical areas and subregions of the hippocampus and efficiently prevents neuronal death in the cortex of hypoglycemic animals. In vitro results show that D-BHB stimulates ATP production and reduces ROS levels, while the nonphysiologic isomer of BHB, L-BHB, has no effect on energy production but reduces ROS levels. Data suggest that protection by BHB, not only results from its metabolic action but is also related to its capability to reduce ROS, rendering this KB as a suitable candidate for the treatment of ischemic and traumatic injury. INTRODUCTIONGlucose is the main energy source in brain. However, under certain conditions other energy substrates such as the ketone bodies (KB), acetoacetate (AcAc), and β-hydroxybutyrate (BHB) can be used by brain, including the suckling period, 1,2 prolonged fasting, 3 and the ketogenic diet; 4 all these situations are associated with increased KB levels in blood. Several studies have shown that KB can protect the brain against damage associated with diverse neurotoxic insults such as hypoxia, 5 ischemia, 6-8 hypoglycemia, 9,10 and excitotoxicity. 11,12 In addition, the ketogenic diet has been used for a long time for the treatment of refractory epilepsy. 4,13,14 The protective effect of KB has been mostly attributed to their conversion to AcetylCoA improving mitochondrial metabolism and preserving energy levels. [15][16][17] However, studies suggest that besides providing energy to neurons, KB reduce the production of reactive oxygen species (ROS) contributing to their protective effect. It has been observed in vitro that AcAc reduces ROS levels induced by glutamate exposure and glycolysis inhibition in cultured neurons, 12,16 and previous in vivo studies have shown that BHB decreases hypoglycemia and glutamate-mediated lipoperoxidation in the rat brain. 10,18 The mechanism underlying the reduction of ROS by BHB has not been elucidated but we have previously reported that KB display a scavenging action of ROS, in particular of the hydoxyl radical ( • OH). 10 The physiologic and the

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“…A decrease in MTT reduction correlates well with the loss of the mitochondrial membrane potential and the decrease in energy levels [43] and it is used as an index of cell viability [42,44]. In previous experiments, we have shown that cell death induced by glycolysis inhibition or GD as monitored by MTT reduction at later times (24 h after the insult) correlates very well with other methods to measure cell death using fluorescent markers or the release of the cytosolic enzyme lactate dehydrogenase [41,45], validating its use as an index of cell viability. In brief, after 20- to 23.5-hour recovery periods, cell cultures were incubated with MTT (150 µ M ) for 1 h at 37°C in a 5% CO 2 /95% O 2 -containing atmosphere.…”

Section: Methodsmentioning

confidence: 93%

“…Immediately after, DMEM medium was substituted for the medium previously withdrawn from each well (Neurobasal glucose-containing medium) and cells were left to recover for 23.5, 23, 22 and 20 h, according to the GD period. Cell viability was monitored by the MTT [3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide] reduction assay [40], as previously described [41]. This assay is based on the ability of living mitochondria to reduce MTT to the insoluble formazan salt by active dehydrogenases [42].…”

Section: Methodsmentioning

confidence: 99%

Neuroprotective Role of Estradiol against Neuronal Death Induced by Glucose Deprivation in Cultured Rat Hippocampal Neurons

et al. 2011

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Studies have reported the protective effect of estradiol (E₂) against neuronal death induced by several insults including oxygen deprivation, mitochondrial toxins and activation of glutamate receptors. Glucose deprivation (GD) is associated with ischemia and hypoglycemia, and to date there is no effective therapeutic agent able to prevent neuronal damage induced by these conditions. In this study, we have investigated the effects of 17β-E₂ and the selective agonists of the alpha (ERα) and beta (ERβ) estrogen receptors, propyl pyrazole triol (PPT) and diarylpropionitrile (DPN), respectively, on neuronal death induced by GD in cultured rat hippocampal neurons. We have also analyzed the expression of both ER isoforms after GD. Results show that GD for 2 and 4 h reduces cell survival by 42 and 55%, respectively. Treatment with 17β-E₂ (10 nM to 10 µM) induces a dose-dependent protective effect that is blocked by ICI 182,780, an ER antagonist, and by 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(-piperidinylethoxy)phenol]-1H′pyrazole dihydrochloride (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP), selective ERα and ERβ antagonists, respectively. The ERα and ERβ agonists PPT and DPN show a similar neuroprotective effect to that of 17β-E₂, but DPN is more efficient. In addition, hippocampal neurons under normal conditions show a higher expression of the ERβ isoform. When exposed to GD during 4 h, the expression of both ER isoforms is increased, while only that of the ERβ isoform significantly increases after 2 h of GD. Results demonstrate that E₂ prevents neuronal death induced by GD through its interaction with ER, although the ERβ isoform might have a predominant role. Results also suggest that GD differentially alters the expression of ERα and ERβ in hippocampal neurons.

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Pathways involved in the generation of reactive oxygen and nitrogen species during glucose deprivation and its role on the death of cultured hippocampal neurons

Cited by 42 publications

References 47 publications

Targeting of multiple metabolites in neural cells monitored by using protein-based carbon nanotubes

Targeting of multiple metabolites in neural cells monitored by using protein-based carbon nanotubes

Protection of Hypoglycemia-Induced Neuronal Death by β-Hydroxybutyrate Involves the Preservation of Energy Levels and Decreased Production of Reactive Oxygen Species

Neuroprotective Role of Estradiol against Neuronal Death Induced by Glucose Deprivation in Cultured Rat Hippocampal Neurons

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