April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Comparative Analysis Of Biological Processes And Molecular Functions In The Human Ocular Epithelium: Preferential Gene Expression In The Human Limbus
Author Affiliations & Notes
  • Martin N. Nakatsu
    Ophthalmology, Jules Stein Eye Institute, Los Angeles, California
  • Daniel M. Vu
    Ophthalmology, Jules Stein Eye Institute, Los Angeles, California
  • Ascia Eskin
    Human Genetics, University of California, Los Angeles, Los Angeles, California
  • Sophie X. Deng
    Ophthalmology, Jules Stein Eye Institute, Los Angeles, California
  • Footnotes
    Commercial Relationships  Martin N. Nakatsu, None; Daniel M. Vu, None; Ascia Eskin, None; Sophie X. Deng, None
  • Footnotes
    Support  Emily G. Plumb Estate and Trust and NEI 5T32EY007026-35
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 2581. doi:
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      Martin N. Nakatsu, Daniel M. Vu, Ascia Eskin, Sophie X. Deng; Comparative Analysis Of Biological Processes And Molecular Functions In The Human Ocular Epithelium: Preferential Gene Expression In The Human Limbus. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2581.

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Abstract

Purpose: : To identify transcriptional differences in gene expression and cell processes in the human limbal epithelium by comparing directly to its adjacent structures, the cornea and conjunctiva.

Methods: : Total RNA was isolated from the limbus, cornea and conjunctiva from 3 separate donors. The gene expression profile of each tissue type was analyzed through a human genome U133 Plus 2.0 microarray and candidate genes from each tissue were identified by expression levels two-fold or higher when compared to each other from the average expression levels of the 3 donors. Selected differentially expressed genes were confirmed by quantitative real time PCR (QRT-PCR) and functional analysis of overexpressed genes in the limbus was performed by the web-based toolkit Database for Annotation, Visualization and Integrated Discovery (DAVID).

Results: : There were 67 differentially expressed (two-fold or higher) transcripts in the limbus in direct comparison to that in the cornea and conjunctiva. Expression of signature markers for the cornea, conjunctiva and limbus were present in their respective tissues in the microarray and confirmed by QRT-PCR. Several genes were differentially expressed in the limbus. They include: tenascin C (TNC), endomucin, neurotrophic tyrosine kinase receptor type 3 (NTRK3), paired-like homeodomain 2 (PITX2), fibromodulin, frizzled-7, Kruppel-like factor 15 (KLF15), axin-2 and angiopoietin-like 7. Functional analysis of the limbal group of overexpressed genes generated several classes involved in various biological processes and cell signaling, including cell-cell adhesion, Wnt signaling, TGF-β signaling and other developmental processes.

Conclusions: : These results demonstrate the potential role for differentially expressed limbal genes in the identification of new limbal epithelial stem cell markers and their involvement in the regulation and maintenance of these cells.

Keywords: gene microarray 
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