title: Gene expression in donor corneal endothelium.
abstract: 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 F(0) 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.
authors.affiliation: Center for Corneal Genetics, The Wilmer Eye Institute, The Johns Hopkins Hospital, 600 N Wolfe St, Baltimore, MD 21287, USA. jgottsch@jhmi.edu
authors.forename: John D
authors.initials: JD
authors.lastname: Gottsch
authors.forename: Gerami D
authors.initials: GD
authors.lastname: Seitzman
authors.forename: Elliott H
authors.initials: EH
authors.lastname: Margulies
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Bowers
authors.forename: Allison J
authors.initials: AJ
authors.lastname: Michels
authors.forename: Saurabh
authors.initials: S
authors.lastname: Saha
authors.forename: Albert S
authors.initials: AS
authors.lastname: Jun
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Stark
authors.forename: Sammy H
authors.initials: SH
authors.lastname: Liu
mesh_terms: D000368:Aged; D018076:DNA, Complementary / Q000235:genetics; D004728:Endothelium, Corneal / Q000737:chemistry*; D020224:Expressed Sequence Tags; D005136:Eye Proteins / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000302:isolation & purification; D016324:Sequence Tagged Sites; D014019:Tissue Donors
chemical_list: D018076:DNA, Complementary; D005136:Eye Proteins; D012313:RNA