Abstract
Purpose: :
Many transcription factors and other proteins that control crucial fate decisions, such as those occurring in the differentiation of specific retinal neuron types, undergo SUMO and/or ubiquitin post-translational modifications. The aim of this work is to analyze the expression of a battery of enzymes involved in the SUMO and ubiquitin conjugation/deconjugation pathways in the mouse retina.
Methods: :
Wild-type retinas from C57BL/6J mice have been the source of material, abiding to the ARVO statement for the use of animals in ophthalmic and vision research. After RNA purification and cDNA obtention, the expression of the battery of genes selected for the study has been analyzed by real-time RT-PCR, in situ hybridization and immunohistochemistry on retinal sections.
Results: :
The genes of the SUMO and ubiquitin pathways analyzed have been classified in categories, according to their level of transcription in the mouse retina. In situ hybridizations performed on retinal sections have shown whether the expression was basal and ubiquitous throughout the retinal cell layers, or whether a specific pattern of expression is indicative of specific neuronal type function for each gene. Those genes with a particularly interesting function or pattern of expression have been further characterized by immunohistochemistry.
Conclusions: :
An expression map for the battery of selected genes encoding SUMO and ubiquitin pathways has been drawn. A subset of genes show specific expression patterns, with higher transcriptional levels in specific cell types, such as the outer and inner plexiform, ganglion cell and photoreceptor cell layers. Future work will focus on the search for specific substrates for the most relevant genes of these two pathways.
Keywords: gene/expression • protein modifications-post translational • retina