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M. Agudo, A. Harrison, S. Mayor–Torroglosa, P. Sobrado, S.B. McMahon, V. Heidinger, A. Doelemeyer, F. Hallböök, J. Miralles, M. Vidal–Sanz; Gene Expression Analysis of Axotomy–Induced Retinal Ganglion Cell Death . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3086.
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Purpose: We have designed a time course experiment using a high–throughput gene expression analysis in an attempt to find out the molecular cause(s) triggering the axotomy–related retinal ganglion cell (RGC) death. Methods: We selected 3 time points after intraorbital optic nerve axotomy where 10% (early), 50% (intermediate) or 80% (late) of RGCs are prompt to die and a control with intact optic nerve. Ten adult rats per time point were subjected to surgery and axomoty was carried out at close distance to the eye. Retinas were quickly extracted and processed for RNA extraction and preparation for microarray hybridization (Affymetrix: rae 230.2). Replicas were done and data collected. Results: First analysis shows RNA expression profiles aligned among replicates. With a cut off of two fold change, approximately 1,056 genes had their expression modified after axotomy, 622 increased (up to 64 fold) and 434 decreased (down to non–detectable) compared to control. Out of the total, 161 genes were regulated at all time points studied. Looking at post–axotomy regulated genes, we have 235 early, 253 intermediate and 210 late changed genes. Besides, 133 genes are regulated early and intermediate but not late, and 74 intermediate and late but not early. Changes were observed in transcription and growth factors (mostly early and intermediate response), cytoskeleton proteins (predominantly late response), immune–response (all times), stress (all times), extracellular matrix, cell cycle (mainly early response), enzymes and unknown genes (all times). Some of these regulated genes, as crystallins, have been described to change also after other insults such as glaucoma, ischemia and retina scraping. Conclusions: Microarray technology allows to differentially study the complex genetic regulation underlying the distinct phases of response to axotomy. Studies performed by other groups using different models of insult, point to a common response to injury in the retina.
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