Microarray analysis of gene expression in human donor corneas

A. S. Jun, S. H. Liu, E. H. Koo, D. V. Do, W. J. Stark, J. D. Gottsch

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Objectives: To use microarray analysis to identify genes expressed in human donor corneas and to create a preliminary, comprehensive database of human corneal gene expression. Methods: A complementary DNA (cDNA) library was constructed from transplant-quality, human donor corneas. Biotin-labeled RNA was transcribed from the cDNA library and hybridized in duplicate to microarrays containing approximately 5600 human genes. Results were analyzed using a gene database of the National Institutes of Health, Bethesda, Md. Reverse transcriptase polymerase chain reaction analysis was performed to confirm corneal expression of genes identified by microarray analysis. Results: Duplicate microarrays identified the expression of 1200 genes in human donor corneas. Chromosomal loci had been assigned to 1025 (85%) of these genes. A preliminary database of human corneal gene expression was compiled. A Web site containing these genes was created. Six collagen genes were identified that had not previously been localized within the cornea. Five apoptosis-related genes were identified, 4 of which had not previously been localized within the cornea. Three genes previously shown to cause corneal diseases were identified. Reverse transcriptase polymerase chain reaction analysis of genes identified by microarray analysis confirmed the corneal expression of 2 apoptosis-related genes and 1 collagen gene. Conclusions: Microarray analysis of healthy human donor corneas has produced a preliminary, comprehensive database of corneal gene expression. Large-scale analysis of gene expression has the potential to generate large amounts of data, which should be made readily accessible to the scientific community. The Internet offers many potential advantages as a medium for the maintenance of these large data sets. Clinical Relevance: Identification of structural, apoptosis-related, and disease-causing genes within the cornea by microarrays may increase the understanding of normal and abnormal corneal function with likely relevance to corneal diseases and transplants.

Original languageEnglish (US)
Pages (from-to)1629-1634
Number of pages6
JournalArchives of Ophthalmology
Volume119
Issue number11
DOIs
StatePublished - 2001

ASJC Scopus subject areas

  • Ophthalmology

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    Jun, A. S., Liu, S. H., Koo, E. H., Do, D. V., Stark, W. J., & Gottsch, J. D. (2001). Microarray analysis of gene expression in human donor corneas. Archives of Ophthalmology, 119(11), 1629-1634. https://doi.org/10.1001/archopht.119.11.1629