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JD Gottsch, A Bowers, EH Margulies, GD Seitzman, SW Kim, S Saha, AS Jun, WJ Stark, SH Liu; Serial Analysis of Gene Expression in the Corneal Endothelium in Fuchs' Dystrophy . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4007.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:To compare the gene expression profiles of normal human corneal endothelium with Fuchs’ corneal endothelium 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’ corneas were stripped and total RNA was isolated. Serial analysis of gene expression (SAGE) analysis was performed to identify and quantify gene transcripts. Genes over- and under-expressed 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 up-regulated and 36 transcripts down- regulated in Fuchs’ endothelium as compared to normal. Up-regulated transcripts included serum amyloid A1 and A2, metallothionein, and apolipoprotein D. Of the down-regulated transcripts, 26 matched known genes, 3 matched ESTs, and 7 were unknown to current databases. One down-regulated 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 Pi, and heat shock 70 kD protein. Nine different SAGE tags matching mitochondrial sequences accounted for 25% of the expressed transcripts that were decreased in Fuchs’ endothelium. Conclusion:SAGE analysis comparing normal to Fuchs’ endothelium demonstrates diminished expression of mitochondrial, pump function, and anti-apoptotic cell defense transcripts. :
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