title: Construction of human corneal endothelial cDNA library and identification of novel active genes.
abstract: OBJECTIVE
  To describe genes expressed in human corneal endothelial cells and identify novel genes.
  
  
  METHODS
  Sixteen human donor corneas that had no history of corneal disease, infection, or intraocular surgery were used within 7 days of death. Total RNA was extracted from corneal endothelial cells with attached Descemet membranes. A 3'-directed cDNA library was constructed from mRNA by using a pUC19-based primer. These sequences were compared with each other to determine their frequency and were searched against GenBank for identification. To identify novel specific and abundant transcript genes in corneal endothelial cells, the novel genes were compared with an expressed sequence tag database, the expected sequence extended, and 5' rapid amplification of cDNA ends-polymerase chain reaction cloning performed.
  
  
  RESULTS
  The human corneal endothelial cDNA library showed that the most abundant transcript was prostaglandin D2 synthase. The remaining transcript genes that were present in abundance consisted of lactate dehydrogenase-A, gene signature (GS) 3582, which is a novel gene without a known function, and matrix Gla protein. The full-length sequence of GS3582 showed similarity to genes obtained in ovary and TESTIS.
  
  
  CONCLUSIONS
  A human corneal endothelial cDNA library was constructed. An expression profile of corneal endothelium provides probes to monitor physiologic and pathologic conditions of this tissue in terms of gene expression.
authors.affiliation: Department of Ophthalmology, Jichi Medical School, Kawachi-gun, Tochigi, Japan.
authors.forename: Rieko
authors.initials: R
authors.lastname: Sakai
authors.forename: Tadatoshi
authors.initials: T
authors.lastname: Kinouchi
authors.forename: Shoko
authors.initials: S
authors.lastname: Kawamoto
authors.forename: M Reza
authors.initials: MR
authors.lastname: Dana
authors.forename: Toshirou
authors.initials: T
authors.lastname: Hamamoto
authors.forename: Tadahiko
authors.initials: T
authors.lastname: Tsuru
authors.forename: Kousaku
authors.initials: K
authors.lastname: Okubo
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
mesh_terms: D000368:Aged; D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000378:metabolism*; D004728:Endothelium, Corneal / Q000378:metabolism*; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D015723:Gene Library; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000302:isolation & purification; D014019:Tissue Donors
chemical_list: D018076:DNA, Complementary; D005136:Eye Proteins; D012313:RNA