Aim: Hyperoxia-induced photoreceptor degeneration occurs preferentially in the inferior retina of C57BL/6 J mice. This study investigates differential gene expression in the inferior and superior retina of C57BL/6 J mouse, before and after hyperoxic stress.
Methods: At the age of P (postnatal day) 83–90, mice were placed in constant normoxia or hyperoxia (75% O2) for 2 weeks. Retinas from control and exposed mice were removed and RNA was extracted from superior and inferior regions. The RNA from 2 animals (1 male and 1 female) at each condition was extracted, purified and hybridized to an Affymetrix MouseGene 1.0 ST Array to elucidate gene expression. Experiments were run in triplicate and analysis of the expression patterns was performed using GeneSpring and Partek Genomics Suite softwares.
Results: Over 400 genes showed significant differential expression by location and treatment using 2-way ANOVA analysis. In the control material, no genes showed a differential expression greater than twofold between inferior and superior retina. After hyperoxic stress, 154 genes in the inferior and 30 genes in the superior retina showed a greater than twofold change in expression. Among those, genes such as Edn2, GFAP, Bcl3 and C1qb showed expression differences of greater than three fold between inferior and superior retina. Real time PCR was used to verify gene expression of control genes as well as genes of interest.
Conclusion: These microarray data may provide clues for identifying previously unknown factors and pathways responsible for the vulnerability of inferior retina to hyperoxic stress and for the eventual identification of therapeutic targets.
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access
This research was supported by grants from the National Health and Medical Research Council (NHMRC) and Australian Research Council (ARC). The authors thank Dr. Stephem Ohms for his assistance in using Partek Genomics software and providing suggestions in statistics.
Chung HS, Harris A, Halter PJ et al (1999) Regional differences in retinal vascular reactivity. Invest Ophthalmol Vis Sci 40:2448–2453PubMedGoogle Scholar
Geller S, Krowka R, Valter K et al (2006) Toxicity of hyperoxia to the retina: evidence from the mouse. Adv Exp Med Biol 572:425–437CrossRefPubMedGoogle Scholar
Natoli R, Provis J, Valter K et al (2008a) Expression and role of the early-response gene Oxr1 in the hyperoxia-challenged mouse retina. Invest Ophthalmol Vis Sci 49:4561–4567CrossRefPubMedGoogle Scholar
Natoli R, Provis J, Valter K et al (2008b) Gene regulation induced in the C57BL/6 J mouse retina by hyperoxia: a temporal microarray study. Mol Vis 14:1983–1994PubMedGoogle Scholar
Noell WK (1955) Visual cell effects of high oxygen pressures. Am Physiol Soc Fed Proc 14:107–108Google Scholar
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45CrossRefPubMedGoogle Scholar
Rapp LM, Williams TP (1977) Rhodopsin content and electroretinographic sensitivity in light-damaged rat retina. Nature 267:835–836CrossRefPubMedGoogle Scholar
Smit-McBride Z, Oltjen SL, LaVail MM et al (2007) A strong genetic determinant of hyperoxia-related retinal degeneration on mouse chromosome 6. Invest Ophthalmol Visual Sci 48:405–411CrossRefGoogle Scholar
Stone J, Maslim J, Valter-Kocsi K et al (1999) Mechanisms of photoreceptor death and survival in mammalian retina. Prog Ret Eye Res 18:689–735CrossRefGoogle Scholar
Walsh N, Bravo-Nuevo A, Geller S et al (2004) Resistance of photoreceptors in the C57BL/6-c2J, C57BL/6 J, and BALB/cJ mouse strains to oxygen stress: evidence of an oxygen phenotype. Curr Eye Res 29:441–447CrossRefPubMedGoogle Scholar
Yamada H, Yamada E, Hackett SF et al (1999) Hyperoxia causes decreased expression of vascular endothelial growth factor and endothelial cell apoptosis in adult retina. J Cell Physiol 179:149–156CrossRefPubMedGoogle Scholar
Yamada H, Yamada E, Ando A et al (2001) Fibroblast growth factor-2 decreases hyperoxia-induced photoreceptor cell death in mice. Am J Pathol 159:1113–1120PubMedGoogle Scholar