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M. Yamasaki, T. Inaba, K. Kashiwagi, K. Murata, Y. Tsumamoto, K. Okada, H. Yamashita, H.K. Mishima; Erythropoietin Prevents Glutamate-induced Retinal Ganglion Cell Death . Invest. Ophthalmol. Vis. Sci. 2003;44(13):5209.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Erythropoietin (EPO) has been reported to protect not only erythroid progenitors but also neurons from apoptosis. We tested the potential of EPO to prevent retinal ganglion cells (RGCs) from glutamate-induced cell death using the primary culture system of RGCs. Methods and Results: RGCs were isolated from 3 to 5 day-old neonatal rats with Thy-1.1 antibody-coated dishes. More than 90% cells obtained by this procedure were proved to be RGC by DiI, a retrograde fluorescent tracer, which was injected subdurally at superior colliculi two days before isolation of RGCs. Expression of the EPO receptor gene in these cells was confirmed using RT-PCR. After incubation for 12 hours in serum-free medium containing EPO (10pM~1nM) or BDNF (4~40 µ), cells were continued to be cultured for 72-hour in medium containing glutamate (25~100 µM), and living cells were evaluated with XTT assay. Glutamate decreased the survival rate of RGCs in a dose-dependent fashion, and EPO as well as BDNF significantly reduced glutamate-induced cell death. Conclusions: EPO has neuroprotective effects on RGCs. Molecular mechanisms through which EPO protects RGCs are now under study.
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