May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Effect of Imposed Defocus on the Profile of Gene Expression in Chick Retinal Pigment Epithelium
Author Affiliations & Notes
  • Z. Zhang
    Vision Science, School of Optometry, University of California, Berkeley, California
  • S. Geller
    Dept. Molecular & Cell Biology, Helen Wills Neuroscience Institute, University of California, Berkeley, California
  • J. Su
    Vision Science, School of Optometry, University of California, Berkeley, California
  • J. G. Flannery
    Vision Science, School of Optometry, University of California, Berkeley, California
    Dept. Molecular & Cell Biology, Helen Wills Neuroscience Institute, University of California, Berkeley, California
  • C. F. Wildsoet
    Vision Science, School of Optometry, University of California, Berkeley, California
  • Footnotes
    Commercial Relationships Z. Zhang, None; S. Geller, None; J. Su, None; J.G. Flannery, None; C.F. Wildsoet, None.
  • Footnotes
    Support NEI Grants EY 12392 (CFW), EY013533 (JGF).
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 4417. doi:
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      Z. Zhang, S. Geller, J. Su, J. G. Flannery, C. F. Wildsoet; Effect of Imposed Defocus on the Profile of Gene Expression in Chick Retinal Pigment Epithelium. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4417.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose:: To elucidate the effect of imposed defocus on the profile of gene expression in chicken retinal pigment epithelium (RPE) using DNA microarray analysis and real-time PCR.

Methods:: White Leghorn chicks wore monocular positive or negative lenses (+10 D & -10 D) from age 19 days for 2 or 48 hours; fellow eyes were untreated. Choroidal thickness responses were measured using high frequency A-scan ultrasonography (n=32). Total RNA was extracted from isolated RPE and used for DNA microarray (n=3) and real-time PCR (n=16) studies. The former used only 48 hr samples and an Affymetrix GeneChip, and data were analyzed using GeneSifter software.

Results:: Out of a possible maximum of 38535 genes detectible with the Affymetrix Chicken Genome chip, 13389 were expressed in RPE from -10 D lens-treated eyes compared to 15881 genes for fellow (control) eyes, and of these, 1582 were exclusive to RPE from control eyes and 431 were exclusive to RPE from eyes wearing -10D lens (n=3, p<0.05). Among the 9992 genes expressed in RPE from both -10D lens-treated and fellow eyes, 157 genes were up-regulated by more than 3 fold and 632 were down-regulated by more than 3 folds in treated compared to fellow eyes. Among the down-regulated genes were TGF-beta3, SMAD2 and SMAD3 (X 2.1, 1.9 and 1.5 fold, respectively). Among the up-regulated genes were TGF-beta receptor1, TGF-beta receptor3, somatostatin, insulin receptor substrate and insulin-like growth factor binding protein (X 2.7, 2.8, 2.1, 2, 2.1 fold, respectively). Real-time PCR analyses revealed up-regulation of somatostatin receptor2 and IGF1R in RPE from -10D lens treated eyes, and down-regulation in RPE from +10D lens-treated eyes at both time points (p<0.05 in all cases). These changes were coupled to significant decreases and increases in choroidal thickness respectively (p<0.01, 2 & 48 hr).

Conclusions:: Imposed defocus results in significant differences in RPE gene expression profiles that also may show sign-dependent differences. These findings offer a mechanism by which the RPE could encode bidirectional retina-derived growth regulatory signals that are assumed to modulate the growth of the choroid and sclera in young eyes. They also provide new directions for research into myopia control therapy.

Keywords: retinal pigment epithelium • myopia • gene microarray 
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