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R De Iongh, JW McAvoy; Microarray Analysis of Gene Expression In Transgenic Lenses Expressing Dominant-Negative TGFß Receptors . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3547.
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Purpose: In previous studies we showed that expression of dominant-negative forms of either type I or type II TGFß receptors in the lens, so as to inhibit TGFß signaling, resulted in degeneration of cortical and nuclear lens fibers (de Iongh et al., 2001, Development 128: 3995-4010.). In one line of mice (OVE591), the phenotype develops between postnatal days 1 and 3. In this study we investigated the effects of inhibiting TGFß signaling on gene expression in these lenses. Methods: Lens fibers were isolated from the lenses of transgenic (OVE591) and wild-type (FVB) mice on postnatal day1 (pre-phenotype) and 3 (post-phenotype). RNA was isolated and reverse transcribed to double-stranded complementary DNA using T7-oligo-dT primers. Labelled cRNA was generated from the cDNA by in vitro transcription, using T7 RNA polymerase and hybridized to Affymetrix GeneChips (Mu11kA) representing a subset of the mouse genome (∼6,500 genes). Results: Gene chip analysis of RNA isolated from wild-type lenses showed that approximately 43% of genes represented on the chips were expressed. Comparison of lenses from P1 transgenic and wild-type mice showed differential expression of 35 genes (11 up-regulated, 24 down-regulated). Up- and down-regulated genes included several cytoskeletal proteins such as tropomyosin-4, cortactin, calpactin, and ß-actin. Analysis of RNA isolated from P3 wild-type and transgenic mice showed differential expression of 142 genes (98 up-regulated, 44 down-regulated). Differentially regulated genes included those associated with apoptosis/cell cycle, intracellular transport, transcription factors and lipid metabolism regulators. Conclusion: These results indicate that during terminal differentiation of lens fibers TGFß signaling plays a major role in modulating expression of cytoskeletal proteins. Disruption of TGFß signaling in lens fibers results in disruption of the cytoskeleton, intracellular transport processes and lipid metabolism and activation of apoptotic pathways.
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