title: Serial analysis of gene expression (SAGE) in normal human trabecular meshwork.
abstract: OBJECTIVE
  To identify the genes expressed in normal human trabecular meshwork tissue, a tissue critical to the pathogenesis of glaucoma.
  
  
  METHODS
  Total RNA was extracted from human trabecular meshwork (HTM) harvested from 3 different donors. Extracted RNA was used to synthesize individual SAGE (serial analysis of gene expression) libraries using the I-SAGE Long kit from Invitrogen. Libraries were analyzed using SAGE 2000 software to extract the 17 base pair sequence tags. The extracted sequence tags were mapped to the genome using SAGE Genie map.
  
  
  RESULTS
  A total of 298,834 SAGE tags were identified from all HTM libraries (96,842, 88,126, and 113,866 tags, respectively). Collectively, there were 107,325 unique tags. There were 10,329 unique tags with a minimum of 2 counts from a single library. These tags were mapped to known unique Unigene clusters. Approximately 29% of the tags (orphan tags) did not map to a known Unigene cluster. Thirteen percent of the tags mapped to at least 2 Unigene clusters. Sequence tags from many glaucoma-related genes, including myocilin, optineurin, and WD repeat domain 36, were identified.
  
  
  CONCLUSIONS
  This is the first time SAGE analysis has been used to characterize the gene expression profile in normal HTM. SAGE analysis provides an unbiased sampling of gene expression of the target tissue. These data will provide new and valuable information to improve understanding of the biology of human aqueous outflow.
authors.affiliation: Center for Human Genetics, Duke University Medical Center, Durham, NC, USA.
authors.forename: Yutao
authors.initials: Y
authors.lastname: Liu
authors.forename: Drew
authors.initials: D
authors.lastname: Munro
authors.forename: David
authors.initials: D
authors.lastname: Layfield
authors.forename: Andrew
authors.initials: A
authors.lastname: Dellinger
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Walter
authors.forename: Katherine
authors.initials: K
authors.lastname: Peterson
authors.forename: Catherine Bowes
authors.initials: CB
authors.lastname: Rickman
authors.forename: R Rand
authors.initials: RR
authors.lastname: Allingham
authors.forename: Michael A
authors.initials: MA
authors.lastname: Hauser
mesh_terms: D000328:Adult; D000368:Aged; D018797:Cell Cycle Proteins; D002874:Chromosome Mapping / Q000379:methods*; D016000:Cluster Analysis; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D020224:Expressed Sequence Tags / Q000737:chemistry*; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods; D015723:Gene Library; D016678:Genome; D005901:Glaucoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D026901:Membrane Transport Proteins; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014129:Trabecular Meshwork / Q000166:cytology / Q000378:metabolism*; D035582:Transcription Factor TFIIIA / Q000235:genetics / Q000378:metabolism; D000099179:Myocilin
chemical_list: D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D006023:Glycoproteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D035582:Transcription Factor TFIIIA; D000099179:Myocilin; C409460:OPTN protein, human; C502341:WDR36 protein, human