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M.H. Sanchez, M.A. Rivas, S.E. Pearce–Kelling, G.M. Acland, D. Rodriguez, G.K. Aguirre, G.D. Aguirre, E. Vecino; Altered Expression of Molecular Markers in the Retina of the LCA–RPE65 Dog Model . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1035.
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© ARVO (1962-2015); The Authors (2016-present)
To characterize the expression of molecular markers specific for different retinal cell types and to analyze specific differences associated with the RP65 mutation.
Retinas from wt and RPE 65 –/– dogs were fixed in paraformaldehyde, embedded in OCT, sectioned at 15 µm and then processed for immunofluorescence. The different cell types were examined using antibodies specific for various molecules. Photoreceptor cells were labeled with antibodies to M/L opsin, S opsin and rod opsin; bipolar cells were labeled with antibodies to Goα or PKC; horizontal and amacrine cells were identified with antibodies to Calcium Binding Proteins; amacrine cells with anti BDNF (brain derived neurotrophic factor), anti–GABA and anti–tyrosine hydroxylase antibodies; ganglion cells with anti– BDNF and anti–neurofilaments; Müller cells and astrocytes with anti–CRALBP, anti–GFAP, anti–glutamine synthetase, anti–p 75, anti–TrKA and anti–vimentin.
Fluorescent microscopic examination of retinal sections revealed differences between wt and RP65 –/– dog retinas. Thus we found a decrease in the expression of photoreceptor markers, and an increase in the expression of glial markers. Moreover some changes were also found in the synapses of bipolar cells.
Molecular differences between wt and RP65 –/– dog retinas, mainly in photoreceptor and glial cells, have been identified using fluorescent immunohistochemistry.
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