Transcription profiling of human optic nerve astrocytes from Caucasian and African Americans with or without glaucoma
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ABSTRACT: This SuperSeries is composed of the following subset Series:; GSE9939: Gene expression data on human optic nerve head astrocytes in normal Caucasian and African americans; GSE9944: Gene expression data on human optic nerve head astrocytes in Caucasian and African americans with or without glaucoma Experiment Overall Design: Refer to individual Series
Project description:To determine whether optic nerve head astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary culture of astrocytes from African American donors with or without glaucoma, compared to astrocytes from Caucasian American donors with or without glaucoma. Experiment Overall Design: We divided samples into four group: Caucasian American normal, Caucasian American with glaucoma, African American normal and African American with glaucoma. We analyzed data from Affymetrix Human Genome U133A 2.0 and U95 chips.
Project description:To determine whether optic nerve head astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary culture of astrocytes from normal African American donors, compared to astrocytes from normal Caucasian American donors. All donors have no histories of eye disease, diabetes, or chronic CNS disease. Experiment Overall Design: We divided samples into two groups: normal Caucasian American and normal African American. We analyzed data from Affymetrix Human Genome Human Genome U133A and U133A 2.0 chips.
Project description:Gene expression in human optic nerve head (ONH) astrocytes exposed to either 60 mm Hg hydrostatic pressure (HP) or control ambient pressure (CP) was compared using Affymetrix GeneChip microarrays to identify HP-responsive genes. Primary ONH astrocytes from two male Caucasian donors (passage 4) were grown to 75% confluence and were exposed for 6, 24 or 48 h to control ambient pressure (CP6, CP24, CP48) or hydrostatic pressure (HP6, HP24, HP48), or harvested at the beginning of the pressure experiment (CP0). Total RNA was extracted using Qiagen RNeasy columns and converted to biotin-labeled cRNA by standard Affymetrix protocols available at web site http://pathology.wustl.edu/~mgacore/genechip.htm#Preparing. Hybridization of the labeled cRNA to Human Genome U95Av2 chips (Affymetrix) was carried out by using Genechip Instrument System (Affymetrix) at Genechip Core Facility of Washington University. A total of 44 chips were generated and distributed as follows: seven for control pressure (CP) at time 0, four CP at 6 h, seven for hydrostatic pressure (HP) at 6 h, eight for CP at 24 h, eight for HP at 24 h, five for CP at 48 h and five for HP at 48h P. The arrays were washed and stained with streptavidin-phycoerythrin followed by scanning on an Agilent GeneArray Scanner G2500A (Agilent Technologies, Palo Alto, CA), and then scanned by an Affymetrix GeneArray Scanner. Data was analyzed by Affymetrix Microarray Suite (version 5.0), linear regression analysis, and GeneSpring expression Analysis Software (version 6.0, Silicon Genetics). For analysis, the samples were scaled to the same target intensity (1500) to allow comparison of multiple samples, and raw data were normalized to median of each gene across all chips for fold change analysis using GeneSpring.
Project description:To determine whether optic nerve head astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary culture of astrocytes from African American donors with or without glaucoma, compared to astrocytes from Caucasian American donors with or without glaucoma. Keywords: Gene expression profile
Project description:To determine whether optic nerve head astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary culture of astrocytes from normal African American donors, compared to astrocytes from normal Caucasian American donors. All donors have no histories of eye disease, diabetes, or chronic CNS disease. Keywords: Gene expression profile
Project description:The optic nerve is an important tissue in glaucoma and the unmyelinated nerve head region remains an important site of many early neurodegenerative changes. In humans and mice, astrocytes constitute the major glial cell type in the region, and in glaucoma they become reactive, influencing the optic nerve head (ONH) microenvironment and disease outcome. To determine the response of ONH astrocytes in glaucoma, we studied their transcriptional response to an elevation in intraocular pressure (IOP) induced by the microbead occlusion model. We also assessed the response of astrocytes in the more distal myelinated optic nerve proper (ONP). In this experimental model, astrocytes of the optic nerve exhibited a region-specific and temporally distinct response: ONH astrocytes showed very few early transcriptional changes and ONP astrocytes demonstrated substantially larger changes over the course of the experiment.
Project description:Primary cultures of astrocytes from rat optic nerve heads were treated with EGFR ligand, EGF. Two cell lines from two different rat donors were used. The sister cell cultures were set as control and EGF treated groups. Keywords: rat optic nerve head astrocytes