Project description:Proliferative vitreoretinopathy (PVR) was induced in a rat model using intravitreal dispase to study fibrosis- and inflammation-related proteome changes and to evaluate the therapeutic effect of topical 3’,4’-dihydroxyflavonol (DiOHF) eye drops. Retina and vitreous were collected 28 days post-injury from vehicle-, DiOHF- and methotrexate-treated eyes, alongside contralateral controls. In parallel, ARPE-19 cells were stimulated with TGFβ1 + PDGF-BB to model wound-healing responses in vitro, with or without DiOHF. Label-free DIA LC-MS/MS was performed (Orbitrap Exploris 480 for rat tissues; Orbitrap Eclipse for ARPE-19), and data were analyzed with Spectronaut directDIA against UniProt Rattus norvegicus or Homo sapiens canonical databases. The dataset contains the original instrument raw files and processed Spectronaut results for both the rat retina/vitreous and ARPE-19 experiments.
Project description:We used expression quantitative trait locus mapping in the laboratory rat (Rattus norvegicus) to gain a broad perspective of gene regulation in the mammalian eye and to identify genetic variation relevant to human eye disease. Of >31,000 gene probes represented on an Affymetrix expression microarray, 18,976 exhibited sufficient signal for reliable analysis and at least 2-fold variation in expression among 120 F2 rats generated from an SR/JrHsd x SHRSP intercross. Genome-wide linkage analysis with 399 genetic markers revealed significant linkage with at least one marker for 1,300 probes (alpha = 0.001; estimated empirical false discovery rate = 2%). Both contiguous and noncontiguous loci were found to be important in regulating mammalian eye gene expression. We investigated one locus of each type in greater detail and identified putative transcription-altering variations in both cases. We found an inserted cREL binding sequence in the 5' flanking sequence of the Abca4 gene associated with an increased expression level of that gene, and we found a mutation of the gene encoding thyroid hormone receptor beta 2 associated with a decreased expression level of the gene encoding short-wave sensitive opsin (Opn1sw). In addition to these positional studies, we performed a pairwise analysis of gene expression to identify genes that are regulated in a coordinated manner and used this approach to validate two previously undescribed genes involved in the human disease Bardet-Biedl syndrome. These data and analytic approaches can be used to facilitate the discovery of additional genes and regulatory elements involved in human eye disease. Experiment Overall Design: mRNA from whole eye tissue was used to assay gene expression level. Samples were acquired from 120 F2 rats generated from an SR/JrHsd x SHRSP intercross. Each sample was run individually, according to standard Affymetrix protocols, without replicates.
Project description:We used expression quantitative trait locus mapping in the laboratory rat (Rattus norvegicus) to gain a broad perspective of gene regulation in the mammalian eye and to identify genetic variation relevant to human eye disease. Of >31,000 gene probes represented on an Affymetrix expression microarray, 18,976 exhibited sufficient signal for reliable analysis and at least 2-fold variation in expression among 120 F2 rats generated from an SR/JrHsd x SHRSP intercross. Genome-wide linkage analysis with 399 genetic markers revealed significant linkage with at least one marker for 1,300 probes (alpha = 0.001; estimated empirical false discovery rate = 2%). Both contiguous and noncontiguous loci were found to be important in regulating mammalian eye gene expression. We investigated one locus of each type in greater detail and identified putative transcription-altering variations in both cases. We found an inserted cREL binding sequence in the 5' flanking sequence of the Abca4 gene associated with an increased expression level of that gene, and we found a mutation of the gene encoding thyroid hormone receptor beta 2 associated with a decreased expression level of the gene encoding short-wave sensitive opsin (Opn1sw). In addition to these positional studies, we performed a pairwise analysis of gene expression to identify genes that are regulated in a coordinated manner and used this approach to validate two previously undescribed genes involved in the human disease Bardet-Biedl syndrome. These data and analytic approaches can be used to facilitate the discovery of additional genes and regulatory elements involved in human eye disease. Keywords: eQTL, F2 cross
Project description:to understand the consequences of chronic exposure to fluoxetine during postnatal life on global transcriptional changes withing the rat hippocamps Agilent one-color experiment,Organism: Rattus Norvegicus, Agilent-016352 Genotypic designed Custom Rattus Norvegicus 8x15k, Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:We report RNA-Seq experiments of whole eye tissues from A/J, BALB/c, and C57BL/6 background mice. Examine ocular tissue from 3 different background mice that display varying rates of retinal degeneration.