Secreted phospholipase A2 potentiates glutamate-induced calcium increase and cell death in primary neuronal cultures

2002 ◽  
Vol 67 (5) ◽  
pp. 634-645 ◽  
Author(s):  
Mark A. DeCoster ◽  
Gerard Lambeau ◽  
Michel Lazdunski ◽  
Nicolas G. Bazan
Keyword(s):  
Cell Death ◽  
Phospholipase A2 ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures ◽  
Secreted Phospholipase A2

scholarly journals Augmentation of poly(ADP-ribose) polymerase-dependent neuronal cell death by acidosis

2016 ◽  
Vol 37 (6) ◽  
pp. 1982-1993 ◽  
Author(s):  
Jian Zhang ◽  
Xiaoling Li ◽  
Herman Kwansa ◽  
Yun Tai Kim ◽  
Liye Yi ◽  
...  
Keyword(s):  
Cell Death ◽  
Ion Channel ◽  
Focal Cerebral Ischemia ◽  
Infarct Volume ◽  
Neuronal Cell ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures ◽  
Tissue Acidosis ◽  
Nuclear Apoptosis ◽  
Apoptosis Inducing Factor

Tissue acidosis is a key component of cerebral ischemic injury, but its influence on cell death signaling pathways is not well defined. One such pathway is parthanatos, in which oxidative damage to DNA results in activation of poly(ADP-ribose) polymerase and generation of poly(ADP-ribose) polymers that trigger release of mitochondrial apoptosis-inducing factor. In primary neuronal cultures, we first investigated whether acidosis per sé is capable of augmenting parthanatos signaling initiated pharmacologically with the DNA alkylating agent, N-methyl- N′-nitro- N-nitrosoguanidine. Exposure of neurons to medium at pH 6.2 for 4 h after N-methyl- N′-nitro- N-nitrosoguanidine washout increased intracellular calcium and augmented the N-methyl- N′-nitro- N-nitrosoguanidine-evoked increase in poly(ADP-ribose) polymers, nuclear apoptosis-inducing factor , and cell death. The augmented nuclear apoptosis-inducing factor and cell death were blocked by the acid-sensitive ion channel-1a inhibitor, psalmotoxin. In vivo, acute hyperglycemia during transient focal cerebral ischemia augmented tissue acidosis, poly(ADP-ribose) polymers formation, and nuclear apoptosis-inducing factor , which was attenuated by a poly(ADP-ribose) polymerase inhibitor. Infarct volume from hyperglycemic ischemia was decreased in poly(ADP-ribose) polymerase 1-null mice. Collectively, these results demonstrate that acidosis can directly amplify neuronal parthanatos in the absence of ischemia through acid-sensitive ion channel-1a . The results further support parthanatos as one of the mechanisms by which ischemia-associated tissue acidosis augments cell death.

Melatonin is protective in necrotic but not in caspasedependent, free radical‐independent apoptotic neuronal cell death in primary neuronal cultures

2000 ◽  
Vol 14 (12) ◽  
pp. 1814-1824 ◽  
Author(s):  
Christoph Harms ◽  
Marion Lautenschlager ◽  
Alexandra Bergk ◽  
Dorette Freyer ◽  
Markus Weih ◽  
...  
Keyword(s):  
Cell Death ◽  
Free Radical ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures

scholarly journals Tau and Caspase 3 as Targets for Neuroprotection

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Anat Idan-Feldman ◽  
Regina Ostritsky ◽  
Illana Gozes
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Drug Candidate ◽  
Mitochondrial Activity ◽  
Neuronal Cultures ◽  
Tau Hyperphosphorylation ◽  
Protective Effects ◽  
Caspase Inhibitor ◽  
Primary Neuronal Cultures

The peptide drug candidate NAP (davunetide) has demonstrated protective effects in variousin vivoandin vitromodels of neurodegeneration. NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death. Recent studies suggest that caspases may play a role in tau pathology. The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system. Our experimental setup used primary neuronal cultures subjected to oxygen-glucose deprivation (OGD), with and without NAP or caspase inhibitor. Cell viability was assessed by measuring mitochondrial activity (MTS assay), and immunoblots were used for analyzing protein level. It was shown that apoptosis was responsible for all cell death occurring following ischemia, and NAP treatment showed a concentration-dependent protection from cell death. Ischemia caused an increase in the levels of active caspase-3 and hyperphosphorylated tau, both of which were prevented by either NAP or caspase-inhibitor treatment. Our data suggest that, in this model system, caspase activation may be an upstream event to tau hyperphosphorylation, although additional studies will be required to fully elucidate the cascade of events.

Effects of NMDA and ferrous sulfate on oxidation and cell death in primary neuronal cultures

2000 ◽  
Vol 37 (5-6) ◽  
pp. 497-507 ◽  
Author(s):  
Joseph G. Rudolph ◽  
John J. Lemasters ◽  
Fulton T. Crews
Keyword(s):  
Cell Death ◽  
Ferrous Sulfate ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures

scholarly journals Anandamide-induced cell death in primary neuronal cultures: role of calpain and caspase pathways

2004 ◽  
Vol 11 (10) ◽  
pp. 1121-1132 ◽  
Author(s):  
V A Movsesyan ◽  
B A Stoica ◽  
A G Yakovlev ◽  
S M Knoblach ◽  
P M Lea ◽  
...  
Keyword(s):  
Cell Death ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures

scholarly journals Evidence for Apoptosis After Intracerebral Hemorrhage in Rat Striatum

2000 ◽  
Vol 20 (2) ◽  
pp. 396-404 ◽  
Author(s):  
Kohji Matsushita ◽  
Wei Meng ◽  
Xiaoying Wang ◽  
Minoru Asahi ◽  
Kazuko Asahi ◽  
...  
Keyword(s):  
Cell Death ◽  
Intracerebral Hemorrhage ◽  
Autologous Blood ◽  
Rat Striatum ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures ◽  
Western Blots ◽  
Neuronal Nuclei ◽  
Dna Laddering ◽  
Brain Extracts

The overall hypothesis that cell death after intracerebral hemorrhage is mediated in part by apoptotic mechanisms was tested. Intracerebral hemorrhage was induced in rats using stereotactic infusions of 0.5 U of collagenase (1-μL volume) into the striatum. After 24 hours, large numbers of TUNEL-positive stained cells with morphologies suggestive of apoptosis were present in the center and periphery of the hemorrhage. Double staining with Nissl and immunocytochemical labeling with antibodies against neuronal nuclei and glial fibrillary acidic protein suggested that these TUNEL-positive cells were mostly neurons and astrocytes. Electrophoresis of hemorrhagic brain extracts showed evidence of DNA laddering into ∼200-bp fragments. Western blots showed cleavage of the cytosolic caspase substrate gelsolin. The density of TUNEL-positive cells at 24 and 48 hours after hemorrhage was significantly reduced by treatment with the broad-spectrum caspase inhibitor zVADfmk. It was unlikely that apoptotic changes were due to neurotoxicity of injected collagenase because TUNEL-positive cells and DNA laddering were also obtained in an alternative model of hemorrhage where autologous blood was infused into the striatum. Furthermore, equivalent doses of collagenase did not induce cell death in primary neuronal cultures. These results provide initial evidence that apoptotic mechanisms may mediate some of the injury in brain after intracerebral hemorrhage.

scholarly journals Clinically Approved Heterocyclics Act on a Mitochondrial Target and Reduce Stroke-induced Pathology

2004 ◽  
Vol 200 (2) ◽  
pp. 211-222 ◽  
Author(s):  
Irina G. Stavrovskaya ◽  
Malini V. Narayanan ◽  
Wenhua Zhang ◽  
Boris F. Krasnikov ◽  
Jill Heemskerk ◽  
...  
Keyword(s):  
Cell Death ◽  
Bioactive Compounds ◽  
Mitochondrial Permeability Transition ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Glucose Deprivation ◽  
Neuronal Cultures ◽  
Primary Neuronal Cultures ◽  
Structural Class

Substantial evidence indicates that mitochondria are a major checkpoint in several pathways leading to neuronal cell death, but discerning critical propagation stages from downstream consequences has been difficult. The mitochondrial permeability transition (mPT) may be critical in stroke-related injury. To address this hypothesis, identify potential therapeutics, and screen for new uses for established drugs with known toxicity, 1,040 FDA-approved drugs and other bioactive compounds were tested as potential mPT inhibitors. We report the identification of 28 structurally related drugs, including tricyclic antidepressants and antipsychotics, capable of delaying the mPT. Clinically achievable doses of one drug in this general structural class that inhibits mPT, promethazine, were protective in both in vitro and mouse models of stroke. Specifically, promethazine protected primary neuronal cultures subjected to oxygen-glucose deprivation and reduced infarct size and neurological impairment in mice subjected to middle cerebral artery occlusion/reperfusion. These results, in conjunction with new insights provided to older studies, (a) suggest a class of safe, tolerable drugs for stroke and neurodegeneration; (b) provide new tools for understanding mitochondrial roles in neuronal cell death; (c) demonstrate the clinical/experimental value of screening collections of bioactive compounds enriched in clinically available agents; and (d) provide discovery-based evidence that mPT is an essential, causative event in stroke-related injury.

scholarly journals Astrocytes Protect against Isoflurane Neurotoxicity by Buffering pro-brain–derived Neurotrophic Factor

2015 ◽  
Vol 123 (4) ◽  
pp. 810-819 ◽  
Author(s):  
Creed M. Stary ◽  
Xiaoyun Sun ◽  
Rona G. Giffard
Keyword(s):  
Cell Death ◽  
Neurotrophic Factor ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Brain Derived Neurotrophic Factor ◽  
Neurotrophin Receptor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Neuronal Cultures ◽  
P75 Neurotrophin Receptor ◽  
Primary Neuronal Cultures

Abstract Background: Isoflurane induces cell death in neurons undergoing synaptogenesis via increased production of pro-brain–derived neurotrophic factor (proBDNF) and activation of postsynaptic p75 neurotrophin receptor (p75NTR). Astrocytes express p75NTR, but their role in neuronal p75NTR-mediated cell death remains unclear. The authors investigated whether astrocytes have the capacity to buffer increases in proBDNF and protect against isoflurane/p75NTR neurotoxicity. Methods: Cell death was assessed in day in vitro (DIV) 7 mouse primary neuronal cultures alone or in co-culture with age-matched or DIV 21 astrocytes with propidium iodide 24 h after 1 h exposure to 2% isoflurane or recombinant proBDNF. Astrocyte-targeted knockdown of p75NTR in co-culture was achieved with small-interfering RNA and astrocyte-specific transfection reagent and verified with immunofluorescence microscopy. proBDNF levels were assessed by enzyme-linked immunosorbent assay. Each experiment used six to eight replicate cultures/condition and was repeated at least three times. Results: Exposure to isoflurane significantly (P < 0.05) increased neuronal cell death in primary neuronal cultures (1.5 ± 0.7 fold, mean ± SD) but not in co-culture with DIV 7 (1.0 ± 0.5 fold) or DIV 21 astrocytes (1.2 ± 1.2 fold). Exogenous proBDNF dose dependently induced neuronal cell death in both primary neuronal and co-cultures, an effect enhanced by astrocyte p75NTR inhibition. Astrocyte-targeted p75NTR knockdown in co-cultures increased media proBDNF (1.2 ± 0.1 fold) and augmented isoflurane-induced neuronal cell death (3.8 ± 3.1 fold). Conclusions: The presence of astrocytes provides protection to growing neurons by buffering increased levels of proBDNF induced by isoflurane. These findings may hold clinical significance for the neonatal and injured brain where increased levels of proBDNF impair neurogenesis.

Sign in / Sign up

Export Citation Format

Share Document