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The effect of nanoseond laser on murine retinal pigment epithelial cells

ABSTRACT: Ophthalmic laser treatment using short pulse lasers has shown promise as a treatment for certain eye diseases such as Age related macular degeeneration. In this study we treated C57BL/6J mice at 9 months of age with 10 laser spots delivered by the 2RT nanosecond laser. Animal were allowed to recover and were aged a further 3 months (12 months old) before retinal pigment epithelial (RPE) cells were isolated. Totsal RNA samples from laser-treated and control animals were isolated and preamplified using the SMARTSeq v4 low RNA input kit. After library preparation a 50bp single end read was performed at a depth of 19-34 million reads per sample using the Illumina HiSeq. Constructs were mapped to the murine genome and differential expression calculated between the laser-treated and control groups. underwent RNAseq o

ORGANISM(S): Mus musculus

PROVIDER: GSE287768 | GEO | 2025/03/26

REPOSITORIES: GEO

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Project description:Background: To identify and functionally annotate cell type-specific gene expression in the human retinal pigment epithelium (RPE), a key tissue involved in age-related macular degeneration and retinitis pigmentosa. Methodology: RPE, photoreceptor and choroidal cells were isolated from selected freshly frozen healthy human donor eyes using laser microdissection. RNA isolation, amplification and hybridization to 44k microarrays was carried out according to Agilent specifications. Bioinformatics was carried out using Rosetta Resolver, David and Ingenuity software. Principal Findings: Our previous 22k analysis of the RPE transcriptome showed that the RPE has high levels of protein synthesis, strong energy demands, is exposed to high levels of oxidative stress and a variable degree of inflammation. We currently use a complementary new strategy aimed at the identification and functional annotation of RPE-specific expressed transcripts. This strategy takes advantage of the multilayered cellular structure of the retina and overcomes a number of limitations of previous studies. In triplicate, we compared the transcriptomes of RPE, photoreceptor and choroidal cells and we deduced RPE specific expression. We identified at least 114 entries with RPE-specific gene expression. Thirty-nine of these 114 genes also show high expression in the RPE, comparison with the literature showed that 85% of these 39 were previously identified to be expressed in the RPE. In the group of 114 RPE specific genes there was an overrepresentation of genes involved in (membrane) transport, vision and ophthalmic disease. More fundamentally, we found RPE-specific involvement in the RAR-activation, retinol metabolism and GABA receptor signaling pathways. Conclusions: In this study we provide a further specification and understanding of the RPE transcriptome by identifying and analyzing genes that are specifically expressed in the RPE. Six RPE samples, three phot samples, three choroid samples

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The effect of nanoseond laser on murine retinal pigment epithelial cells

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