Gene expression and functional studies of the optic nerve head astrocyte transcriptome from normal African Americans and Caucasian Americans donors.

PloS One
Haixi MiaoM R Hernandez

Abstract

To determine whether optic nerve head (ONH) astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary cultures of astrocytes from normal African American (AA) donors compared to astrocytes from normal Caucasian American (CA) donors. We used oligonucleotide Affymetrix microarray (HG U133A & HG U133A 2.0 chips) to compare gene expression levels in cultured ONH astrocytes from twelve CA and twelve AA normal age matched donor eyes. Chips were normalized with Robust Microarray Analysis (RMA) in R using Bioconductor. Significant differential gene expression levels were detected using mixed effects modeling and Statistical Analysis of Microarray (SAM). Functional analysis and Gene Ontology were used to classify differentially expressed genes. Differential gene expression was validated by quantitative real time RT-PCR. Protein levels were detected by Western blots and ELISA. Cell adhesion and migration assays tested physiological responses. Glutathione (GSH) assay detected levels of intracellular GSH. Multiple analyses selected 87 genes differentially expressed between normal AA and CA (P<0.01). The most relevant genes expressed in AA were categorized by function, including: signal...Continue Reading

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Citations

Dec 21, 2010·Nature Neuroscience·Alex BayésSeth G N Grant
Jul 26, 2011·Current Eye Research·Wei XueVijay P Sarthy
Jun 7, 2011·Experimental Eye Research·Abe Clark
Sep 1, 2015·Experimental Eye Research·Jan Christoph Koch, Paul Lingor
Aug 17, 2010·Experimental Eye Research·Gülgün Tezel
Jun 21, 2015·Experimental Eye Research·Hee Joo ChoiTatjana C Jakobs

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Methods Mentioned

BETA
chips
Chip
ELISA
GTPases
glucosylation
electrophoresis
in vitro transcription
Protein Assay
Assay
PCR

Software Mentioned

Limma
R
Lima
GOstats Bioconductor
RMA
Bioconductor
SAM

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