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1 Departments of Biological Chemistry and Ophthalmology, University of California, Davis, California, USA; Vitreoretinal Research Laboratory, School of Medicine University of California, Davis, California, USA
* To whom correspondence should be addressed. E-mail: lmhjelmeland{at}ucdavis.edu.
To evaluate the age-related changes in gene expression occurring in the complex of retinal pigmented epithelium, Bruch's membrane, and choroid (RPE/choroid), we examined the gene expression profiles of young adult (2-month) and old (24-month) male C57BL/6 mice. cDNA probe sets from individual animals were synthesized using total RNA isolated from the RPE/choroid of each animal. Probes were amplified using the Clontech SMART system, radioactively labeled, and hybridized to two different Clontech Atlas mouse cDNA arrays. From each age group, three independent triplicates were hybridized to the arrays. Statistical analyses were performed using the Significance Analysis of Microarrays program (SAM Version 1.13; Stanford University). Selected array results were confirmed by semi-quantitative RT-PCR analysis. Out of 2340 genes represented on the arrays, approximately 60% were expressed in young and/or old mouse RPE/choroid. A moderate fraction (12%) of all expressed genes exhibited a statistically significant change in expression with age. Of these 150 genes, all but two, HMG14 and carboxypeptidsase E, were up-regulated with age. Many of these up-regulated genes can be grouped into several broad functional categories: immune response, proteases and protease inhibitors, stress response, and neovascularization. RT-PCR results from six out of six genes examined confirmed the differential change in expression with age of these genes. Our study provides likely candidate genes to further study their role in the development of age-related macular degeneration and other aging diseases affecting the RPE/choroid.
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