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L. Panagis, X. Zhao, L. Ren, X. Yang, J. Danias, T. W. Mittag; Gene Expression Changes in Areas of Focal Loss of Retinal Ganglion Cells (RGC) in the Retina of DBA/2J Mice. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5488.
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To determine whether differences in gene expression occur between areas of focal RGC loss and areas of relative RGC preservation in the retina of the DBA/2J mouse model of glaucoma.
Adult, 11 -13 months old, DBA/2J mice of (N=10) were used. To determine the amount of ganglion cell loss, retinas were pre-labeled retrogradely with FluoroGold The animals were sacrificed, the eyes were enucleated and the retinas were flat mounted, examined and for focal RGC loss. Areas of focal loss and adjacent areas with relative RGC preservation were dissected and stored in RNA-later. RNA was extracted and subjected to microarray analysis using Affymetrix oligonucleotide arrays. RT-PCR was performed to confirm some of the genes identified to be differentially expressed.
Microarray analysis revealed that expression of 372 genes was up regulated while expression of 119 genes was down regulated by 0.5 fold in areas of RGC loss. Genes identified to be differentially expressed include genes coding for cytoskeletal proteins, enzymes, transport proteins, as well as extracellular matrix proteins and immune response proteins. Ten of the genes identified were confirmed by RT-PCR.
Regional gene expression changes occur in retinas with areas of focal RGC loss and areas of preservation in DBA/2J mice. Genes involved code for proteins with diverse cellular functions. Further investigation is required to determine the cellular localisation of the expression changes of these genes during the development of spontaneous glaucoma in the DBA/2J mouse and to determine whether these gene expression changes are causative or protective of RGC loss.
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