Abstract
Abstract: :
Purpose: To examine the protective effect of donepezil, an acetylcholinesterase inhibitor which was developed as a therapeutic agent for Alzheimer’s disease, in cultured retinal ganglion cells (RGCs). Methods: RGCs were purified from dissociated rat retinal cells (postnatal day 6–8), using a modified two–step panning method and cultured in serum–free medium containing neurotrophic factors and forskolin. Survival of RGCs after exposure to glutamate (Glu, 25µM), with or without donepezil (Dnp, 100nM–100µM) was measured by calcein–acetoxymethyl ester staining after 3 days of exposure. Surviving RGCs were defined as cells with a calcein–stained cell body and a process extending at least two cell diameters from the cell body. Results: The application of Glu caused RGC death. Simultaneous application of Dnp reduced glutamate–induced RGC death in a dose–dependent manner. (Survival ratio: Glu only vs. Glu+Dnp (100µM), 61.7±9.4% vs. 91.7±14.5%, mean±SD n=9, p<0.001) Conclusions: These findings suggest that donepezil may inhibit glutamate–induced cell death in purified RGCs.
Keywords: ganglion cells • retinal culture • neuroprotection