Expression profile and genome location of cDNA clones from an infant human trabecular meshwork cell library.
OBJECTIVE
To delineate the profile of genes expressed in infant human trabecular meshwork and identify candidate genes for glaucoma.


METHODS
Human trabecular meshwork cell cultures were established from six young donors. A cDNA library was made from the combined trabecular meshwork mRNA. The end-sequence of random clones was determined by direct sequencing. These sequences were then analyzed by a National Center for Biotechnology Information (NCBI, Bethesda, MD) database search. Nucleotide searches were performed using the BLASTN (ver. 2.1.3; against the nonredundant nucleic acid sequence) and dbEST databases (both provided by NCBI in the public domain at www.ncbi.nlm.nih.gov).


RESULTS
Sequences from 1118 clones from this nonamplified trabecular meshwork cDNA library were categorized. Of these, 877 expressed sequence tags (ESTs) (78.7%) were known genes. One hundred thirty-nine ESTs (12.5%) showed close identity to EST sequences reported in the public domain database (dbEST). Thirteen ESTs (1.2%) showed no significant similarity to known genes or ESTs in the public databases and were thus defined as novel ESTs. The most abundant genes expressed by the human trabecular meshwork included ferritin H, eukaryotic translation elongation factor 1-alpha, ferritin L, fibronectin, and TIMP-1. Ferritin H was the most abundant transcript, making up more than 4% of the genes expressed by the human trabecular meshwork. Extracellular matrix proteins were also highly expressed. The chromosome location of the trabecular meshwork ESTs is reported.


CONCLUSIONS
A profile of genes expressed by human trabecular meshwork is presented. Thirteen novel ESTs were identified. The combined information obtained from expression analysis and chromosomal localization of trabecular meshwork cDNAs should be valuable in identifying candidate genes for glaucoma.
Wirtz
Mary K
MK
Department of Ophthalmology, Casey Eye Institute, Oregon Health and Sciences University, Portland 97201, USA. wirtzm@ohsu.edu
Samples
John R
JR
Xu
Hong
H
Severson
Tesa
T
Acott
Ted S
TS
D002478:Cells, Cultured; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D015723:Gene Library; D016678:Genome; D005901:Glaucoma / Q000235:genetics; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014129:Trabecular Meshwork / Q000166:cytology / Q000378:metabolism*
D005136:Eye Proteins; D012333:RNA, Messenger