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Eri Kubo, Shinsuke Shibata, Naoko Shibata, Teppei Shibata, Etsuko Kiyokawa, Hiroshi Sasaki, Dhirendra P Singh; Gene expression profiling of lens epithelial cells in a rat posterior capsular opacity model in vivo. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2646.
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© ARVO (1962-2015); The Authors (2016-present)
The epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) has been proposed as a major cause of posterior capsule opacification (PCO) after cataract surgery. Using an in vivo rodent PCO model, we earlier demonstrated that the expression of tropomyosin (Tpm)1α/2β was aberrantly upregulated in remodeling the actin cytoskeleton of LECs. Using a microarray-based approach, herein we studied the changes in gene expression pattern during rat PCO formation in vivo as a model.
All animal experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Extracapsular lens extractions (ECLEs) were performed in Sprague Dawley rats to generate the rat PCO model. To identify differential gene expression patterns during PCO, LECs with capsules were isolated at Day 0 (control), 1week (W), and 2W following ECLEs. Total RNA was extracted from the lenses and labelled cDNA was hybridized to Agilent SurePrint G3 Rat 8x60K microarray. Changes in gene expression level were analyzed. Real-time PCR and western blot analysis were used to validate the microarray results.
PCOs were formed following 1W and 2W of ECLE. Expression of several genes such as decorin, a member of the small leucine-rich proteoglycan family, collagen (type 1, 3, 5, 6) and fibronectin were upregulated (3-6 fold) in rat LECs compared to those of the control. The rat LECs also showed increased expression of TGFβ- inducible gene h3 and Tpm2 mRNA and protein were expressed compared to control (Day 0). Expressions of γ-crystalline and filensin were higher in LECs isolated at 2W compared to the control and 1W after ECLE.
These findings spotted the stage specific gene expression patterns during PCO formation. The identifying regulation of genes linked to EMT or fiber differentiation in rat PCO models may provide strategies to develop stage-specific therapeutic agents to block progression of PCO.
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