RETRACTED: Characterization of a transformed rat retinal ganglion cell line

Krishnamoorthy, Raghu R; Agarwal, Puneet; Prasanna, Ganesh; Vopat, K.; Lambert, Wendi S.; Sheedlo, Harold J.; Pang, Iok‐Hou; Shade, Debra L.; Wordinger, Robert J.; Yorio, Thomas; Clark, Abbot F.; Agarwal, Neeraj

doi:10.1016/s0169-328x(00)00224-2

Cited by 294 publications

(251 citation statements)

References 44 publications

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“…It has been reported that the RGC-5 cell line expresses many proteins characteristic of ganglion cells, such as THY-1, Brn 3C and NMDA-receptor [21] . As described in the original publication, when the cells are not differentiated in culture, they do not demonstrate the characteristic susceptibility to glutamate that intact ganglion cells demonstrate.…”

Section: Discussionmentioning

confidence: 99%

“…The development of the rat ganglion cell line (RGC-5) has been described [21]. RGC-5 cells were maintained at 37°C in a humidified chamber of 5% CO 2.…”

Section: Cell Culturementioning

confidence: 99%

“…The differentiation of RGC-5 cells was promoted following the method of Krishnamoorthy et al [21]. Briefly, RGC-5 cells, which had been grown as described above, were cultured in the absence of serum for 24 h, after which they were cultured in DMEM (4 mM glutamine, 1% FBS, 100 U/ml penicillin, 100 µg/ml streptomycin) supplemented with 50 µg/ml succinyl concanavalin A (sConA) for 7 days.…”

Section: Homocysteine Excitotoxicity In Cultured Rgcsmentioning

confidence: 99%

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The sigma receptor ligand (+)−pentazocine prevents apoptotic retinal ganglion cell death induced in vitro by homocysteine and glutamate

Martin

Ola

Agarwal

et al. 2004

Molecular Brain Research

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Recent studies demonstrated that the excitotoxic amino acid homocysteine induces apoptotic death of retinal ganglion cells in vivo. In the present study, an in vitro rat retinal ganglion cell culture system was used to analyze the toxicity of acute exposure to high levels of homocysteine, the mechanism of homocysteine-induced toxicity and the usefulness of σR1 ligands as neuroprotectants. When cultured RGC-5 cells were subjected to treatment with 1 mM D, Lhomocysteine, a significant increase in cell death was detected by TUNEL analysis and analysis of activated caspase. When cells were treated with homocysteine-or glutamate in the presence of MK-801, an antagonist of the NMDA receptor, the cell death was inhibited significantly. In contrast, NBQX, an antagonist of the AMPA/Kainate receptor, and nifedipine, a calcium channel blocker, did not prevent the homocysteine-or glutamate-induced cell death. Semi-quantitative RT-PCR and immunocytochemical analysis demonstrated that RGC-5 cells exposed to homocysteine or glutamate express type 1 sigma receptor at levels similar to control cells. Treatment of RGC-5 cells with 3 µM or 10 µM concentrations of the σR1-specific ligand (+)-pentazocine inhibited significantly the apoptotic cell death induced by homocysteine or glutamate. The results suggest that homocysteine is toxic to ganglion cells in vitro, that the toxicity is mediated via NMDA receptor activation, and that the σR1-specific ligand (+)-pentazocine can block the RGC-5 cell death induced by homocysteine and glutamate.

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Section: Discussionmentioning

confidence: 99%

“…The development of the rat ganglion cell line (RGC-5) has been described [21]. RGC-5 cells were maintained at 37°C in a humidified chamber of 5% CO 2.…”

Section: Cell Culturementioning

confidence: 99%

Section: Homocysteine Excitotoxicity In Cultured Rgcsmentioning

confidence: 99%

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The sigma receptor ligand (+)−pentazocine prevents apoptotic retinal ganglion cell death induced in vitro by homocysteine and glutamate

Martin

Ola

Agarwal

et al. 2004

Molecular Brain Research

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“…The development of the rat retinal ganglion cell line (RGC-5) has been described [27,18]. These cells express markers for ganglion cells such as Thy-1, Brn-3C, neurotrophins and their cognate receptors, but they are negative for markers of Müller, amacrine, and horizontal cells suggesting they represent a ganglion cells.…”

Section: Culture Of Rgc-5 Cellsmentioning

confidence: 99%

“…In the present study, we used the retinal ganglion cell line (RGC-5) [27] to assess the effects of various NO donors on the activity and expression of the system x c -. Since this is a transformed cell line, we also studied primary ganglion cells isolated from neonatal mouse retinas and determined that xCT was indeed present in these primary neurons.…”

Section: Introductionmentioning

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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In Situ Forming Hydrogels: A Thermo‐Responsive Polyelectrolyte as Promising Liquid Artificial Vitreous Body Replacement

et al. 2011

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The use of fully synthetic polymers as a liquid artificial vitreous body replacement (LAVR) was already considered, but yet none of these polymers have proven to be successful in long term retention in the vitreous cavity due to inadequate biocompatibility and fast biodegradation. Our novel cross‐linked polyelectrolyte as a potential intraocular endotamponade shows an innovative concept of thermoresponsive behavior proposed for the first time in ophthalmologic surgery. The polyelectrolyte has been elaborated via a three‐step procedure by combining classical synthetic protocols and microwave technique. We observed that the polyelectrolyte comprising sodium 2‐acrylamido‐2‐methylpropane sulfonate and acrylic functionalized poly(N‐isopropylacrylamide) macromonomer with glycerol dimethacrylate as cross‐linker displays a noticeable thermosensitive behavior. It exhibits enhancement of shear viscosity at physiological temperatures without phase separation. First in vitro experiments on a retinal ganglion cell line (RGC‐5) present a promising biocompatibility without harmful effects. Cell viability, proliferation rate, and inflammatory cascade (COX‐2 and iNOS expression) were not affected.

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RETRACTED: Characterization of a transformed rat retinal ganglion cell line

Cited by 294 publications

References 44 publications

The sigma receptor ligand (+)−pentazocine prevents apoptotic retinal ganglion cell death induced in vitro by homocysteine and glutamate

The sigma receptor ligand (+)−pentazocine prevents apoptotic retinal ganglion cell death induced in vitro by homocysteine and glutamate

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

In Situ Forming Hydrogels: A Thermo‐Responsive Polyelectrolyte as Promising Liquid Artificial Vitreous Body Replacement

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