Microarray reveals complement components are regulated in the serum-deprived rat retinal ganglion cell line

Glaucoma is a progressive eye disease that leads to blindness due to loss of retinal ganglion cells (RGCs). There are difficulties in using primary cultures of purified RGC to study this pathophysiology. RGC-5, a transformed not RGC line, expresses several markers characteristic of the RGCs. The aim...

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Bibliographic Details
Published in:Molecular vision 2007-02, Vol.13, p.293-308
Main Authors: Khalyfa, Abdelnaby, Chlon, Timothy, Qiang, He, Agarwal, Neeraj, Cooper, Nigel G F
Format: Article
Language:English
Subjects:
Animals
Apoptosis - genetics
Cell Line, Transformed
Complement C1 - metabolism
Complement C3 - metabolism
Complement Factor H - metabolism
Complement System Proteins - metabolism
Computer Systems
Culture Media, Serum-Free
Down-Regulation
Gene Expression Profiling
Gene Expression Regulation
Immunohistochemistry
Microscopy, Confocal
Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction
Rats
Reproducibility of Results
Retinal Ganglion Cells - metabolism
Signal Transduction - genetics
Tissue Distribution
Up-Regulation
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Summary:Glaucoma is a progressive eye disease that leads to blindness due to loss of retinal ganglion cells (RGCs). There are difficulties in using primary cultures of purified RGC to study this pathophysiology. RGC-5, a transformed not RGC line, expresses several markers characteristic of the RGCs. The aim of this study was to generate a genome-wide gene expression of RGC-5 following serum deprivation and to identify candidate genes that may be involved in the signal transduction pathways. Apoptosis in the transformed rat RGC-5 was induced by serum deprivation for 0, 8, 24, 48, and 96 h. Briefly, 400 ng of RNA from each sample was reverse transcribed and labeled with Cy3 dye. Fragmented fluorescent cRNA was mixed with hybridization buffer and incubated at 60 degrees C for 16 h. Labeled cRNA was hybridized to Rat Genome Oligonucleotide Arrays. These arrays contain 22,775 transcripts with one oligonucleotide per transcript (60-mer). Gene expression from scanned images was quantified and analyzed using ArrayVision software. Reproducibility among triplicate arrays was determined by ANOVA statistical analysis. Significant differences in gene expression between apoptotic and nonapoptotic cells were determined based on p-values. Of the 22,775 transcripts present on the arrays (Agilent rat genome, 60-mer), 713 (8 h), 1,967 (24 h), 1,011 (48 h), and 1,161 (96 h) were differentially expressed relative to the 0 h time point (p-values
ISSN:1090-0535