Pigment epithelium-derived factor protects cultured retinal neurons against hydrogen peroxide-induced cell death

W. Cao, Joyce Tombran-Tink, W. Chen, D. Mrazek, R. Elias, J. F. McGinnis

Research output: Contribution to journalArticle

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Abstract

Pigment epithelium-derived factor (PEDF) is a neurotrophic protein synthesized and secreted by retinal pigment epithelial (RPE) cells in early embryogenesis and has been shown to be present in the extracellular matrix between the RPE cells and the neural retina. It induces neuronal differentiation and promotes survival of neurons of the central nervous system from degeneration caused by serum withdrawal or glutamate cytotoxicity. Because the role of PEDF in the retina is still unknown, we examined its ability to protect cultured retinal neurons against hydrogen peroxide (H2O2)-induced cell death. Retinas of 0-2-day-old Sprague-Dawley rats were isolated and dissociated, and the neurons were maintained for 2 weeks in a synthetic serum-free medium. Immunocytochemical labeling showed that 50-60% of the cultured cells were rod photoreceptors. Treatment with H2O2 induced significant death of retinal neurons in a dose- and time- dependent manner. Pretreatment with PEDF prior to insult greatly attenuated H2O2-induced cytotoxicity, and its effect was shown to be dose dependent. Cytotoxicity was determined by 3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl- tetrazolium bromide and lactate dehydrogenase assays, and apoptotic cell death was evaluated by the TdT-mediated digoxigenin-dUTP nick-end labeling assay. The present study also showed that H2O2-induced retinal neuron death was by apoptosis that could be inhibited by PEDF. Combination of PEDF with basic fibroblast growth factor, brain-derived neurotrophic factor, or ciliary neurotrophic factor improves the protection. These data strongly suggest that PEDF is a potential neuroprotective agent in the treatment of retinal degeneration.

Original languageEnglish (US)
Pages (from-to)789-800
Number of pages12
JournalJournal of Neuroscience Research
Volume57
Issue number6
DOIs
StatePublished - Sep 15 1999

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Retinal Neurons
Hydrogen Peroxide
Cell Death
Retina
Retinal Pigments
Epithelial Cells
Ciliary Neurotrophic Factor
Retinal Rod Photoreceptor Cells
Neurons
Digoxigenin
Retinal Degeneration
Brain-Derived Neurotrophic Factor
Serum-Free Culture Media
Nerve Growth Factors
Neuroprotective Agents
Fibroblast Growth Factor 2
L-Lactate Dehydrogenase
Embryonic Development
Extracellular Matrix
Sprague Dawley Rats

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience

Cite this

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title = "Pigment epithelium-derived factor protects cultured retinal neurons against hydrogen peroxide-induced cell death",
abstract = "Pigment epithelium-derived factor (PEDF) is a neurotrophic protein synthesized and secreted by retinal pigment epithelial (RPE) cells in early embryogenesis and has been shown to be present in the extracellular matrix between the RPE cells and the neural retina. It induces neuronal differentiation and promotes survival of neurons of the central nervous system from degeneration caused by serum withdrawal or glutamate cytotoxicity. Because the role of PEDF in the retina is still unknown, we examined its ability to protect cultured retinal neurons against hydrogen peroxide (H2O2)-induced cell death. Retinas of 0-2-day-old Sprague-Dawley rats were isolated and dissociated, and the neurons were maintained for 2 weeks in a synthetic serum-free medium. Immunocytochemical labeling showed that 50-60{\%} of the cultured cells were rod photoreceptors. Treatment with H2O2 induced significant death of retinal neurons in a dose- and time- dependent manner. Pretreatment with PEDF prior to insult greatly attenuated H2O2-induced cytotoxicity, and its effect was shown to be dose dependent. Cytotoxicity was determined by 3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl- tetrazolium bromide and lactate dehydrogenase assays, and apoptotic cell death was evaluated by the TdT-mediated digoxigenin-dUTP nick-end labeling assay. The present study also showed that H2O2-induced retinal neuron death was by apoptosis that could be inhibited by PEDF. Combination of PEDF with basic fibroblast growth factor, brain-derived neurotrophic factor, or ciliary neurotrophic factor improves the protection. These data strongly suggest that PEDF is a potential neuroprotective agent in the treatment of retinal degeneration.",
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Pigment epithelium-derived factor protects cultured retinal neurons against hydrogen peroxide-induced cell death. / Cao, W.; Tombran-Tink, Joyce; Chen, W.; Mrazek, D.; Elias, R.; McGinnis, J. F.

In: Journal of Neuroscience Research, Vol. 57, No. 6, 15.09.1999, p. 789-800.

Research output: Contribution to journalArticle

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T1 - Pigment epithelium-derived factor protects cultured retinal neurons against hydrogen peroxide-induced cell death

AU - Cao, W.

AU - Tombran-Tink, Joyce

AU - Chen, W.

AU - Mrazek, D.

AU - Elias, R.

AU - McGinnis, J. F.

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N2 - Pigment epithelium-derived factor (PEDF) is a neurotrophic protein synthesized and secreted by retinal pigment epithelial (RPE) cells in early embryogenesis and has been shown to be present in the extracellular matrix between the RPE cells and the neural retina. It induces neuronal differentiation and promotes survival of neurons of the central nervous system from degeneration caused by serum withdrawal or glutamate cytotoxicity. Because the role of PEDF in the retina is still unknown, we examined its ability to protect cultured retinal neurons against hydrogen peroxide (H2O2)-induced cell death. Retinas of 0-2-day-old Sprague-Dawley rats were isolated and dissociated, and the neurons were maintained for 2 weeks in a synthetic serum-free medium. Immunocytochemical labeling showed that 50-60% of the cultured cells were rod photoreceptors. Treatment with H2O2 induced significant death of retinal neurons in a dose- and time- dependent manner. Pretreatment with PEDF prior to insult greatly attenuated H2O2-induced cytotoxicity, and its effect was shown to be dose dependent. Cytotoxicity was determined by 3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl- tetrazolium bromide and lactate dehydrogenase assays, and apoptotic cell death was evaluated by the TdT-mediated digoxigenin-dUTP nick-end labeling assay. The present study also showed that H2O2-induced retinal neuron death was by apoptosis that could be inhibited by PEDF. Combination of PEDF with basic fibroblast growth factor, brain-derived neurotrophic factor, or ciliary neurotrophic factor improves the protection. These data strongly suggest that PEDF is a potential neuroprotective agent in the treatment of retinal degeneration.

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