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John D. Gottsch, Amanda L. Bowers, Elliott H. Margulies, Gerami D. Seitzman, Sean W. Kim, Saurabh Saha, Albert S. Jun, Walter J. Stark, Sammy H. Liu; Serial Analysis of Gene Expression in the Corneal Endothelium of Fuchs’ Dystrophy. Invest. Ophthalmol. Vis. Sci. 2003;44(2):594-599. doi: 10.1167/iovs.02-0300.
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purpose. To compare the gene expression profiles of normal human corneal endothelium with Fuchs’ corneal endothelium, by using serial analysis of gene expression (SAGE).
methods. Three pairs of normal human corneas were obtained from eye banks. Thirteen bisected Fuchs’ corneal buttons were processed at the time of corneal transplantation. The endothelia of normal and Fuchs’-affected corneas were stripped, and total RNA was isolated. Serial analysis of gene expression (SAGE) was performed to identify and quantify gene transcripts. Genes over- and underexpressed by Fuchs’ endothelium were limited to P < 0.01 by the method of Audic and Claverie.
results. A total of 19,136 tags were identified with 9,530 from normal and 9,606 from Fuchs’ endothelium. The expression of 18 transcripts was upregulated, and 36 transcripts were downregulated in Fuchs’ endothelium compared with normal tissue. Upregulated transcripts included serum amyloid A1 and A2, metallothionein, and apolipoprotein D. Of the downregulated transcripts, 26 matched known genes, 3 matched expressed sequence tags (ESTs), and 7 were unknown to current databases. One downregulated transcript involved a newly reported bicarbonate transporter. Decreased transcripts related to antioxidants and proteins conferring protection against toxic stress were noted in Fuchs’ versus normal endothelium including nuclear ferritin, glutathione S-transferase-π, and heat shock 70-kDa protein. Nine different SAGE tags matching mitochondrial sequences accounted for 25% of the ESTs that were decreased in Fuchs’ endothelium.
conclusions. SAGE analysis comparing normal to Fuchs’ endothelium demonstrates diminished expression of mitochondrial, pump function, and antiapoptotic cell defense genes.
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