Cytoskeletal Breakdown and Apoptosis Elicited by NO Donors in Cerebellar Granule Cells Require NMDA Receptor Activation

Bonfoco, Emanuela; Leist, Marcel; Zhivotovsky, Boris; Orrenius, Sten; Lipton, Stuart A.; Nicotera, Pierluigi

doi:10.1046/j.1471-4159.1996.67062484.x

Cited by 129 publications

(66 citation statements)

References 47 publications

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“…NO is a key factor leading to neuronal death triggered by glutamateinduced stimulation of the NMDA receptor. This has been demonstrated in many neuronal cell types [32][33][34] including retinal neurons [33,35]. In retina, a low concentration of NO plays a protective role in glutamate neurotoxicity by closing the NMDA receptor gated ion channel, but elevated levels of NO, interacting with oxygen radicals, become toxic and mediate glutamate-induced neurotoxicity [35,36].…”

Section: Discussionmentioning

confidence: 99%

Expression of the cystine-glutamate exchanger (xc −) in retinal ganglion cells and regulation by nitric oxide and oxidative stress

et al. 2006

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Purpose-The cystine-glutamate transporter, system x c -, mediates the Na + -independent exchange of cystine into cells coupled to the efflux of intracellular glutamate. Within cells, cystine is reduced to cysteine, a component of glutathione, thus system x c -plays a critical role in glutathione homeostasis. Studies in brain had initially suggested that system x c -was present primarily in astrocytes and not neurons, but more recent work suggests that certain brain neurons have an active system x c -. In the retina, system x c -has been demonstrated in retinal Müller and RPE cells. Recent immunohistochemical work from our lab suggested that the two protein components of system x c -, xCT and 4F2hc, are present in ganglion cells of the intact retina. The purpose of this investigation was to determine whether system x c -was present in primary ganglion cells isolated from neonatal mouse retinas and, if so, to study its regulation by oxidative stress in a retinal ganglion cell line, RGC-5.Methods-The presence of xCT and 4F2hc in RGC-5 cells was established by RT-PCR, immunoblotting and immunohistochemistry and in primary mouse retinal ganglion cells by immunoblotting and immunohistochemistry; the function of the transporter in RGC-5 cells was established by measuring radiolabeled glutamate uptake in the absence of Na + . To assess regulation of system x c -by oxidative stress in ganglion cells, RGC-5 cells were incubated in the presence or absence of nitric oxide (NO) donors (3-nitroso-N-acetylpenicillamine (SNAP), Snitrosoglutathione (SNOG), or 3-morpholinosydnonimine hydrochloride (SIN-1) and reactive oxygen species (ROS) donors menadione sodium bisulfite, hydrogen peroxide and xanthine sodium salt/xanthine oxidase and system x c -was analyzed using functional assays, RT-PCR and immunoblotting.Results-RGC-5 cells and primary ganglion cells isolated from mouse retina express xCT and 4F2hc as demonstrated by RT-PCR, immunoblotting and immunohistochemistry. RGC-5 cells take up glutamate in the absence of Na + and this uptake was blocked by known inhibitors of system x c -, glutamate, cysteine, and cystine as well as quisqualic acid. Treatment of RGC-5 cells with NO donors and donors of ROS led to an increase in the functional activity of system x c -. Kinetic analysis of SNAP-treated RGC-5 cells compared to control cells showed that the increase was associated with an increase in the maximal velocity of the transporter with no significant change in the substrate affinity. Molecular analyses showed that the upregulation is associated with an increase in the expression of xCT with no detectable change in the expression of 4F2hc. Conclusions-RGC-5 cells and ganglion cells isolated from neonatal mice express the cystine/ glutamate transporter x c -(the light chain xCT and the heavy chain 4F2hc) as is evident from functional and molecular studies. Oxidative stress upregulates this transport system in RGC-5 cells and the process is associated with an increase in xCT mRNA and protein but no change in 4F2hc mRNA or...

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Section: Discussionmentioning

confidence: 99%

Expression of the cystine-glutamate exchanger (xc −) in retinal ganglion cells and regulation by nitric oxide and oxidative stress

et al. 2006

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“…1C, red arrows). This morphology has been interpreted as representing apoptosis in some studies examining the effects of low NMDA concentrations (Bonfoco et al, 1995(Bonfoco et al, , 1996 and in past studies of in vitro traumatic neuronal damage (Shah et al, 1997). Moreover, apoptosis has been implicated as a causative death mechanism in many animal studies of TBI (Rink et al, 1995;Clark et al, 1997;Conti et al, 1998;Newcomb et al, 1999;Raghupathi et al, 2000;Wennersten et al, 2003).…”

Section: Sublethal Stretch Injury Produces Irregular Nuclear Morphologymentioning

confidence: 98%

Vulnerability of Central Neurons to Secondary Insults afterIn VitroMechanical Stretch

Arundine¹,

Aarts²,

Lau³

et al. 2004

J. Neurosci.

115

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Mild traumatic brain injuries are of major public health significance. Neurons in such injuries often survive the primary mechanical deformation only to succumb to subsequent insults. To study mechanisms of vulnerability of injured neurons to secondary insults, we used an in vitro model of sublethal mechanical stretch. Stretch enhanced the vulnerability of the neurons to excitotoxic insults, causing nuclear irregularities, DNA fragmentation, and death suggestive of apoptosis. However, the DNA degradation was not attributable to classical (caspase mediated) or caspase-independent apoptosis. Rather, it was associated with profound stretch-induced mitochondrial dysfunction and the overproduction of reactive oxygen species (ROS). Sublethally stretched neurons produced surprisingly high levels of ROS, but these in isolation were insufficient to kill the cells. To be lethal, the ROS also needed to combine with nitric oxide (NO) to form the highly reactive species peroxynitrite. Peroxynitrite was not produced after stretch alone and arose only after combining stretch with an insult capable of stimulating NO production, such as NMDA or an NO donor. This explained the exquisite sensitivity of sublethally stretched neurons to a secondary NMDA insult. ROS scavengers and NO synthase (NOS) inhibitors prevented cell death and DNA degradation. Moreover, inhibiting neuronal NOS activation by NMDA using peptides that perturb NMDA receptor-postsynaptic density-95 interactions also reduced protein nitration and cell death, indicating that the reactive nitrogen species produced were neuronal in origin. Our data explain the mechanism of enhanced vulnerability of sublethally injured neurons to secondary excitotoxic insults and highlight the importance of secondary mechanisms to the ultimate outcome of neurons in mild neurotrauma.

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“…In fact, NO is an extremely versatile messenger in biological systems, and has been implicated in a number of different physiopathological roles, such as smooth muscle relaxation, platelet inhibition, neurotransmission, immune regulation, cell differentiation and tissue morphogenesis (Stamler and Piantadosi, 1996;Stamler et al, 1997;Brune et al, 1999;Nicotera et al, 1999a;Gu et al, 2002) and cytotoxicity (Brune et al, 1999). Cell death elicited by endogenously derived NO plays a central role in pathologic phenomena such as septic shock, nonspecific 'host' defense against tumors and intracellular pathogens, acute and chronic neurodegenerative diseases, pancreatic b-cell destruction in diabetes mellitus and transplant rejection (Choi, 1988;Moncada et al, 1991;Bredt and Snyder, 1994;Kaneto et al, 1995;Bonfoco et al, 1996;Nicotera et al, 1997Nicotera et al, , 1999bLeist and Nicotera, 1998). Figure 3 shows a schematic pattern of the action of NO, from its synthesis, chemical interactions, intracellular biochemical effectors, leading to its biological actions.…”

Section: Energy Requirement For the Shape Of Cell Deathmentioning

confidence: 99%

“…This is mediated by a chemical interaction with mdm2, which fails to degrade and inactivate p53 (Wang et al, 2002(Wang et al, , 2003Schneiderhan et al, 2003). In the excitotoxic death of cultured neurons, NO-triggered apoptosis requires a Ca 2 þ signal triggered by the activation of the NMDAR channels (Bonfoco et al, 1996). Depletion of ATP in PC12 cells, caused via the NO-mediated inhibition of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase, is one of the mechanisms responsible for NO neurotoxicity (Yasuda et al, 1998).…”

Section: No-induced Apoptosismentioning

confidence: 99%

Regulation of the apoptosis–necrosis switch

2004

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Cytoskeletal Breakdown and Apoptosis Elicited by NO Donors in Cerebellar Granule Cells Require NMDA Receptor Activation

Cited by 129 publications

References 47 publications

Expression of the cystine-glutamate exchanger (xc −) in retinal ganglion cells and regulation by nitric oxide and oxidative stress

Expression of the cystine-glutamate exchanger (xc −) in retinal ganglion cells and regulation by nitric oxide and oxidative stress

Vulnerability of Central Neurons to Secondary Insults afterIn VitroMechanical Stretch

Regulation of the apoptosis–necrosis switch

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