scholarly journals Glial Glycogen Stores Affect Neuronal Survival during Glucose Deprivation in vitro

1993 ◽  
Vol 13 (1) ◽  
pp. 162-169 ◽  
Author(s):  
Raymond A. Swanson ◽  
Dennis W. Choi
Keyword(s):  
Neuronal Degeneration ◽  
Neuronal Survival ◽  
Cortical Cell ◽  
Glucose Deprivation ◽  
Methionine Sulfoximine ◽  
Protective Effects ◽  
Cortical Cell Cultures ◽  
Energy Dependent ◽  
Glycogen Stores

Glia perform several energy-dependent functions that may aid neuronal survival under pathological conditions. Glycogen is the major energy reserve in brain, and it is localized almost exclusively to astrocytes. Using murine cortical cell cultures containing both glia and neurons, we examined the effect of altered glial glycogen stores on neuronal survival following glucose deprivation. As previously reported, cultures exposed for several hours to media lacking glucose developed widespread neuronal degeneration without glial degeneration. If glial astrocyte glycogen content was increased to 2–3 times control levels by a 24-h pretreatment with 1 μ M insulin or 0.5 m M methionine sulfoximine (MSO), glucose deprivation-induced neuronal degeneration was attenuated. These protective effects were blocked if glycogen levels were reduced back to control levels by a 30-min exposure to 1 m M dibutyryl cyclic AMP or 20 μ M norepinephrine prior to glucose deprivation. Astrocyte glycogen stores may be an important factor influencing neuronal survival under conditions of energy substrate limitation.

scholarly journals Secobarbital Attenuates Excitotoxicity but Potentiates Oxygen—Glucose Deprivation Neuronal Injury in Cortical Cell Culture

1993 ◽  
Vol 13 (5) ◽  
pp. 803-810 ◽  
Author(s):  
Rona G. Giffard ◽  
John H. Weiss ◽  
Raymond A. Swanson ◽  
Dennis W. Choi
Keyword(s):  
Neuronal Death ◽  
Excitatory Amino Acids ◽  
Cortical Cell ◽  
Glucose Deprivation ◽  
Atp Depletion ◽  
Oxygen Glucose Deprivation ◽  
Protective Effects ◽  
Atp Production ◽  
High Concentrations ◽  
Cortical Cell Cultures

We examined the effects of secobarbital and other sedative-hypnotic barbiturates on the neuronal death induced by exposure to excitatory amino acids or deprivation of oxygen or glucose in mouse cortical cell cultures. N-Methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate, and kainate toxicities were attenuated in a concentration-dependent fashion by high concentrations of secobarbital or thiopental. Antagonism of NMDA toxicity was not overcome by increasing NMDA concentration and not mimicked by γ-aminobutyrate. Despite these antiexcitotoxic actions, secobarbital exacerbated the neuronal death induced by deprivation of either glucose alone or oxygen and glucose together; death induced by oxygen deprivation alone was little affected. Thiopental and methohexital also increased oxygen-glucose deprivation injury. A possible explanation for this injury potentiation was provided by the observation that secobarbital enhanced the cellular ATP depletion induced by combined oxygen-glucose deprivation. Deleterious effects on ATP production may counterbalance the protective effects of barbiturates under some conditions.

Protective effect of nigella sativa extract and thymoquinone on serum/glucose deprivation-induced PC12 cells death

2011 ◽  
Vol 26 (S2) ◽  
pp. 908-908
Author(s):  
H.R. Sadeghnia ◽  
S.H. Mousavi ◽  
Z. Tayarani-Najaran ◽  
M. Asghari
Keyword(s):  
Cell Viability ◽  
Pc12 Cells ◽  
Neurodegenerative Disorders ◽  
Nigella Sativa ◽  
Glucose Deprivation ◽  
Serum Glucose ◽  
Ros Production ◽  
Protective Effects ◽  
Intracellular Ros

The serum/glucose deprivation (SGD)-induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders.Nigella sativa L. and its active component, thymoquinone (TQ) have been known as a source of antioxidants. In the present study, the protective effects of N. sativa and TQ on cell viability and reactive oxygen species (ROS) production in cultured PC12 cells were investigated under SGD conditions. PC12 Cells were pretreated with different concentrations of N. sativa extract (15.62–250 μg/ml) and TQ (1.17–150 μM) for 2 h and then subjected to SGD for 6 or 18 h. Cell viability was quantitated by MTT assay. Intracellular ROS production was measured by flow cytometry using 2’,7’-dichlorofluorescin diacetate (DCF-DA) as a probe. SGD induced significant cells toxicity after 6, 18, or 24 h (p < 0.001). Pretreatment with N. sativa (15.62–250 μg/ml) and TQ (1.17–37.5 μM) reduced SGD-induced cytotoxicity in PC12 cells after 6 and 18 h. A significant increase in intracellular ROS production was seen following SGD (p < 0.001). N. sativa (250 μg/ml, p < 0.01) and TQ (2.34, 4.68, 9.37 μM, p < 0.01) pretreatment reversed the increased ROS production following ischemic insult. The experimental results suggest that N. sativa extract and TQ protects the PC12 cells against SGD-induced cytotoxicity via antioxidant mechanisms. Our findings might raise the possibility of potential therapeutic application of N. sativa extract and TQ for managing cerebral ischemic and neurodegenerative disorders.

Oxygen or glucose deprivation-induced neuronal injury in cortical cell cultures is reduced by tetanus toxin

Neuron ◽  
1992 ◽  
Vol 8 (5) ◽  
pp. 967-973 ◽  
Author(s):  
H. Monyer ◽  
R.G. Giffard ◽  
D.M. Hartley ◽  
L.L. Dugan ◽  
M.P. Goldberg ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Tetanus Toxin ◽  
Cortical Cell ◽  
Neuronal Injury ◽  
Glucose Deprivation ◽  
Cortical Cell Cultures

scholarly journals Gp91phox (NOX2) in Activated Microglia Exacerbates Neuronal Damage Induced by Oxygen Glucose Deprivation and Hyperglycemia in an in Vitro Model

2018 ◽  
Vol 50 (2) ◽  
pp. 783-797 ◽  
Author(s):  
Xianzhang Zeng ◽  
Hongliang Ren ◽  
Yana Zhu ◽  
Ruru Zhang ◽  
Xinxin Xue ◽  
...  
Keyword(s):  
Ischemia Reperfusion Injury ◽  
Neuronal Damage ◽  
Neuronal Survival ◽  
Ischemia Reperfusion ◽  
Survival Rates ◽  
Glucose Deprivation ◽  
Oxygen Glucose Deprivation ◽  
Sirna Transfection ◽  
Culture Model

Background/Aims: Peri-operative cerebral ischemia reperfusion injury is one of the most serious peri-operative complications that can be aggravated in patients with diabetes. A previous study showed that microglia NOX2 (a NADPH oxidase enzyme) may play an important role in this process. Here, we investigated whether increased microglial derived gp91phox, also known as NOX2, reduced oxygen glucose deprivation (OGD) after induction of hyperglycemia (HG). Methods: A rat neuronal-microglial in vitro co-culture model was used to determine the effects of gp91phox knockdown on OGD after HG using six treatment groups: A rat microglia and neuron co-culture model was established and divided into the following six groups: high glucose + scrambled siRNA transfection (HG, n = 5); HG + gp91phoxsiRNA transfection (HG-gp91siRNA, n = 5); oxygen glucose deprivation + scrambled siRNA transfection (OGD, n = 5); OGD + gp91phoxsiRNA transfection (OGD-gp91siRNA, n = 5); HG + OGD + scrambled siRNA transfection (HG-OGD, n = 5); and HG + OGD + gp91phoxsiRNA transfection (HG-OGD-gp91siRNA, n = 5). The neuronal survival rate was measured by the MTT assay, while western blotting was used to determine gp91phox expression. Microglial derived ROS and neuronal apoptosis rates were analyzed by flow cytometry. Finally, the secretion of cytokines, including IL-6, IL-8, TNF-α, and 8-iso-PGF2α was determined using an ELISA kit. Results: Neuronal survival rates were significantly decreased by HG and OGD, while knockdown of gp91phox reversed these rates. ROS production and cytokine secretion were also significantly increased by HG and OGD but were significantly inhibited by knockdown of gp91phoxsiRNA. Conclusion: Knockdown of gp91phoxsiRNA significantly reduced oxidative stress and the inflammatory response, and alleviated neuronal damage after HG and OGD treatment in a rat neuronal-microglial co-culture model.

scholarly journals Dexmedetomidine protects against oxygen-glucose deprivation/reoxygenation injury-induced apoptosis via the p38 MAPK/ERK signalling pathway

2017 ◽  
Vol 46 (2) ◽  
pp. 675-686 ◽  
Author(s):  
Ke Wang ◽  
Yuekun Zhu
Keyword(s):  
Cell Survival ◽  
P38 Mapk ◽  
Glucose Deprivation ◽  
Signalling Pathway ◽  
Cell Counting ◽  
Oxygen Glucose Deprivation ◽  
Protective Effects ◽  
N2a Cells ◽  
Induced Apoptosis

Objective To investigate the protective effects of dexmedetomidine (DEX) in oxygen-glucose deprivation/reoxygenation (OGD/R) injury, which is involved in a number of ischaemic diseases. Methods An in vitro OGD/R injury model was generated using mouse Neuro 2A neuroblastoma (N2A) cells. Different concentrations of DEX were administrated to OGD/R cells. CV-65 was used to inhibit p38 microtubule associated protein kinase/extracellular signal-regulated kinases (MAPK/ERK) signalling. Cell proliferation, cell cycle, apoptosis, and the levels of proteins related to p38 MAPK/ERK signalling and apoptosis were evaluated using Cell Counting Kit-8, flow cytometry, TdT-UTP nick end labelling and Western blot analysis, respectively. Results DEX treatment of OGD/R cells promoted cell survival and attenuated OGD/R-induced cell apoptosis. It also activated the p38 MAPK/ERK signalling pathway, increased the levels of Bcl-2, and decreased the levels of Bax and cleaved caspase-3. Treatment with the p38 MAPK/ERK inhibitor CV-65 inhibited the activation of p38 MAPK/ERK and abrogated the DEX-induced effects on cell survival and apoptosis. Conclusions DEX protects N2A cells from OGD/R-induced apoptosis via the activation of the p38 MAPK/ERK signalling pathway. DEX might be an effective agent for the treatment of ischaemic diseases.

scholarly journals 2,7-Bis-(4-Amidinobenzylidene)-Cycloheptan-1-One Dihydrochloride, tPA Stop, Prevents tPA-Enhanced Excitotoxicity Both In Vitro and In Vivo

2004 ◽  
Vol 24 (10) ◽  
pp. 1153-1159 ◽  
Author(s):  
Géraldine Liot ◽  
Karim Benchenane ◽  
Frédéric Léveillé ◽  
José P. López-Atalaya ◽  
Mónica Fernández-Monreal ◽  
...  
Keyword(s):  
Brain Injuries ◽  
Calcium Influx ◽  
Cortical Cell ◽  
Tissue Type ◽  
Type Plasminogen Activator ◽  
Neuroprotective Strategy ◽  
Cerebral Parenchyma ◽  
Cortical Cell Cultures

Tissue-type plasminogen activator (tPA) is available for the treatment of thromboembolic stroke in humans. However, adverse effects of tPA have been observed in animal models of ischemic brain injuries. In the present study, we have used a synthetic tPA inhibitor, named 2,7-bis-(4-amidinobenzylidene)-cycloheptan-1-one dihydrochloride (tPA stop), to investigate the role of endogenous tPA in the cerebral parenchyma. In mouse cortical cell cultures, we observed that although tPA stop reduced N-methyl-d-aspartic acid (NMDA)-mediated excitotoxic neuronal death, it failed to modulate α-amino-2,3-dihydro-5-methyl-3-oxo-4-isoxazole propanoic acid or kainate-mediated necrosis. In addition, we found that tPA stop could prevent the deleterious effects of both endogenous and exogenous tPA during NMDA exposure. At the functional level, tPA stop was found to prevent tPA-dependent potentiation of NMDA receptor-evoked calcium influx. The relevance of those findings was strengthened by the observation of a massive reduction of NMDA-induced excitotoxic lesion in rats when tPA stop was co-injected. Altogether, these data demonstrate that the blockade of the endogenous proteolytic activity of tPA in the cerebral parenchyma could be a powerful neuroprotective strategy raised against brain pathologies associated with excitotoxicity.

scholarly journals GABAA Receptor Activation Attenuates Excitotoxicity but Exacerbates Oxygen—Glucose Deprivation-Induced Neuronal Injury In Vitro

1996 ◽  
Vol 16 (6) ◽  
pp. 1211-1218 ◽  
Author(s):  
Judith K. Muir ◽  
Doug Lobner ◽  
Hannelore Monyer ◽  
Dennis W. Choi
Keyword(s):  
Neuronal Death ◽  
Receptor Agonist ◽  
Cortical Cell ◽  
Glucose Deprivation ◽  
Receptor Activation ◽  
Oxygen Glucose Deprivation ◽  
Receptor Stimulation ◽  
Bathing Medium

We examined the effects of GABA receptor stimulation on the neuronal death induced by exogenously added excitatory amino acids or combined oxygen–glucose deprivation in mouse cortical cell cultures. Death induced by exposure to NMDA, AMPA, or kainate was attenuated by addition of GABA or the GABAA receptor agonist, muscimol, but not by the GABAB receptor agonist, baclofen. The antiexcitotoxic effect of GABAA receptor agonists was blocked by bicuculline or Picrotoxin. In contrast, GABA or muscimol, but not baclofen, markedly increased the neuronal death induced by oxygen–glucose deprivation. Muscimol potentiation of neuronal death was associated with increased glutamate efflux to the bathing medium, and increased cellular 45Ca2+ accumulation; it was blocked by MK-801, but not NBQX, suggesting mediation by NMDA receptors. Bicuculline only weakly attenuated muscimol potentiation of oxygen–glucose deprivation-induced neuronal death, probably because it itself increased this death. Present results raise a note of caution in the proposed use of GABAA receptor stimulation to limit ischemic brain damage in vivo.

scholarly journals Systematic quantification of synapses in primary neuronal culture

2020 ◽  
Author(s):  
Peter Verstraelen ◽  
Gerardo Garcia ◽  
Marlies Verschuuren ◽  
Bob Asselbergh ◽  
Rony Nuydens ◽  
...  
Keyword(s):  
Synapse Formation ◽  
Dynamic Range ◽  
Cortical Cell ◽  
Neuronal Culture ◽  
Maturity Stage ◽  
Primary Neuronal Culture ◽  
Synaptic Markers ◽  
Synapse Density ◽  
Cortical Cell Cultures

AbstractA vast set of neurological disorders is associated with impaired synaptic connectivity. Therefore, modulation of synapse formation could have therapeutic relevance. However, the high density and small size of synapses make their quantification a challenging task. To improve the reliability of synapse-oriented drug screens, we evaluated a panel of synapse-targeting antibodies for their labeling specificity on hippocampal and cortical cell cultures using quantitative immunofluorescence microscopy. For those antibodies that passed multiparametric validation, we assessed pairwise colocalization, an often-used readout for established synapses. We found that even when two pan-synaptic markers were used, the overlap was incomplete, and the presence of spurious signals limited the dynamic range. To circumvent this problem, we implemented a proximity ligation-based approach, that only leads to a signal when two pre- and postsynaptic markers are sufficiently close. We demonstrate that this approach can be applied to different synaptic marker combinations and can be successfully used for quantification of synapse density in cultures of different maturity stage in healthy or pathological conditions. Thus, the unbiased analysis of synapse labeling and exploitation of resident protein proximity, allows increasing the sensitivity of synapse quantifications in neuronal culture and therefore represents a valuable extension of the analytical toolset for in vitro synapse screens.

scholarly journals Interleukin-1β Protects Neurons against Oxidant-Induced Injury via the Promotion of Astrocyte Glutathione Production

Antioxidants ◽  
2018 ◽  
Vol 7 (8) ◽  
pp. 100 ◽  
Author(s):  
Twinkle Chowdhury ◽  
Matthew Allen ◽  
Trista Thorn ◽  
Yan He ◽  
Sandra Hewett
Keyword(s):  
Neurological Diseases ◽  
Oxidant Stress ◽  
Cortical Cell ◽  
Mixed Cultures ◽  
Interleukin 1Β ◽  
Neuronal Cultures ◽  
Dependent Manner ◽  
Protective Effects ◽  
Oxidant Injury

Interleukin-1β (IL-1β), a key cytokine that drives neuroinflammation in the Central Nervous System (CNS), is enhanced in many neurological diseases/disorders. Although IL-1β contributes to and/or sustains pathophysiological processes in the CNS, we recently demonstrated that IL-1β can protect cortical astrocytes from oxidant injury in a glutathione (GSH)-dependent manner. To test whether IL-1β could similarly protect neurons against oxidant stress, near pure neuronal cultures or mixed cortical cell cultures containing neurons and astrocytes were exposed to the organic peroxide, tert-butyl hydroperoxide (t-BOOH), following treatment with IL-1β or its vehicle. Neurons and astrocytes in mixed cultures, but not pure neurons, were significantly protected from the toxicity of t-BOOH following treatment with IL-1β in association with enhanced GSH production/release. IL-1β failed to increase the GSH levels or to provide protection against t-BOOH toxicity in chimeric mixed cultures consisting of IL-1R1+/+ neurons plated on top of IL-1R1−/− astrocytes. The attenuation of GSH release via block of multidrug resistance-associated protein 1 (MRP1) transport also abrogated the protective effect of IL-1β. These protective effects were not strictly an in vitro phenomenon as we found an increased striatal vulnerability to 3-nitropropionic acid-mediated oxidative stress in IL-1R1 null mice. Overall, our data indicate that IL-1β protects neurons against oxidant injury and that this likely occurs in a non-cell-autonomous manner that relies on an increase in astrocyte GSH production and release.

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