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Gene Expression in Donor Corneal Endothelium
John D. Gottsch, MD;
Gerami D. Seitzman, MD;
Elliott H. Margulies, PhD;
Amanda L. Bowers, BA;
Allison J. Michels, BA;
Saurabh Saha, MD, PhD;
Albert S. Jun, MD, PhD;
Walter J. Stark, MD;
Sammy H. Liu, PhD
Arch Ophthalmol. 2003;121:252-258.
Objective To report gene expression profiles of normal human corneal endothelium with microarray analysis and serial analysis of gene expression (SAGE).
Methods Corneal endothelium was removed from normal human corneas obtained from eye banks. Total RNA was isolated and SAGE analysis was performed. The same RNA source was used to construct a complementary DNA library that was hybridized to microarrays containing 12 558 transcripts.
Results A total of 9530 SAGE tags were sequenced, representing 4724 unique tags. Microarray analysis identified 542 distinct transcripts. A database of human corneal endothelial gene expression was compiled. Of the SAGE tags, 1720 matched known genes, 478 corresponded to expressed sequence tags, and 2526 had no known match to public databases. The 5 most abundantly expressed SAGE tags were cytochrome c oxidase subunit II, adenosine triphosphate synthase F0 subunit 6, carbonic anhydrase XII, 12S ribosomal RNA, and ferritin, heavy polypeptide 1. Thirty-four percent of the transcripts (n = 1616) were specific to the corneal endothelium, when compared with other publicly available SAGE libraries. The 5 most abundant unique tags were keratin 12, angiopoietinlike factor, annexin A8, and 2 tags with no match to the database. Many endothelial pump function enzymes were confirmed, including several plasma membrane Na+/K+ adenosine triphosphatases and a recently reported bicarbonate transporter.
Conclusions Corneal endothelial gene expression profiles by the current analysis provide an understanding of endothelial metabolism, structure, and function; enable comparisons to diseased endothelium; and provide baseline data that may lead to the discovery of novel endothelial genes.
From the Center for Corneal Genetics, Cornea and External Disease Service, The Wilmer Eye Institute (Drs Gottsch, Seitzman, Jun, Stark, and Liu and Mss Bowers and Michels), and Johns Hopkins Oncology Center Molecular Genetics Laboratory, The Johns Hopkins School of Medicine (Dr Saha), Baltimore, Md; and Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Md (Dr Margulies).
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