Abstract
Abstract: :
Purpose: To better understand the gene expression profiles of transformed rat retinal ganglion cells (RGC–5), and to identify candidate genes, which may be involved in apoptotic signal transduction pathways in optic neuropathies including glaucoma. Methods: Apoptosis in the transformed rat retinal ganglion cell line (RGC–5) was induced by serum deprivation for 0, 8, 24, 48, and 96 hours. Fragmented fluorescent cRNA was mixed with hybridization buffer and incubated at 60oC for 16 hours. Labeled cRNA was hybridized to Rat Genome oligoarrays (Agilent). These arrays contain 22,775 transcripts with one oligonucleotide per transcript (60–mer). Reproducibility among triplicates arrays was determined by ANOVA methods. Significant differences in gene expression between apoptotic and non–apoptotic cells was determined based on p–values. Results: Gene expression analysis of RGC–5 cells showed that about 4,620 genes were up–regulated, and 250 down–regulated in serum deprived conditions. After filtration there were about 569 up–regulated and 38 down–regulated genes. The most significantly up–regulated genes were: Gadd45a, Casp11, P2rx4, Scya5, and Dnase1. Highly down–regulated genes were: Pik3e3, Kcns3, Cabp1, Kcnip1, and Hist4.<br /Conclusions: We have compared the expression of 22,775 transcripts of the rat genome using commercial oligonucleotide–based microarrays. The data identified several novel genes in addition to genes that may be important for induction of apoptosis in RGC–5 cells. Supported by NIH:NCRR 2 P20RR016481 (NC) and American Health Assistance Foundation–National Glaucoma Program (NA).
Keywords: cell death/apoptosis • gene microarray • gene/expression