May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Increase in Clusterin Expression in Fuchs' Endothelial Dystrophy
Author Affiliations & Notes
  • U. V. Jurkunas
    Ophthalmology, Harvard Medical School, Massachusetts Eye & Ear Infirmary, Boston, Massachusetts
    Schepens Eye Research Institute, Boston, Massachusetts
  • M. Bitar
    Ophthalmology, Harvard Medical School,
    Schepens Eye Research Institute, Boston, Massachusetts
  • I. Rawe
    Schepens Eye Research Institute, Boston, Massachusetts
  • D. L. Harris
    Ophthalmology, Harvard Medical School,
    Schepens Eye Research Institute, Boston, Massachusetts
  • K. Colby
    Ophthalmology, Harvard Medical School, Massachusetts Eye & Ear Infirmary, Boston, Massachusetts
    Schepens Eye Research Institute, Boston, Massachusetts
  • N. C. Joyce
    Ophthalmology, Harvard Medical School,
    Schepens Eye Research Institute, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  U.V. Jurkunas, None; M. Bitar, None; I. Rawe, None; D.L. Harris, None; K. Colby, None; N.C. Joyce, None.
  • Footnotes
    Support  NEI K12 EY016335
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 1013. doi:https://doi.org/
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    • Get Citation

      U. V. Jurkunas, M. Bitar, I. Rawe, D. L. Harris, K. Colby, N. C. Joyce; Increase in Clusterin Expression in Fuchs' Endothelial Dystrophy. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1013. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To investigate the differential expression of the glycoprotein clusterin/apoJ (CLU) in normal and Fuchs’ endothelial dystrophy (FED) corneal endothelium and to compare the expression of various forms of CLU in normal and FED tissue.

Methods: : FED and pseudophakic bullous keratopathy (PBK) corneal buttons were removed during transplantation and normal corneas were obtained from tissue banks. Human corneal endothelial cells and Descemet’s membrane (HCEC-DM) complex was dissected from the stroma. Proteins were separated on 2-D gels and subjected to comparative proteomic analysis. Relative expression of pre-secretory CLU (pre-sCLU), secretory CLU (sCLU), and nuclear CLU (nCLU) were compared between normal and FED HCEC-DM by Western blots. Expression of CLU mRNA was compared using RT-PCR. Subcellular localization of CLU was compared in corneal whole mounts from normal and FED patients by immunocytochemistry followed by confocal microscopy.

Results: : Proteomic analysis revealed an apparent increase in CLU expression in FED HCEC-DM compared with normal controls. Western blot analysis demonstrated that pre-sCLU protein expression was 4.5 times higher in FED than in normal samples (p=0.002), while the mature form modified for secretion (sCLU) was not significantly elevated (p=0.29). Expression of nCLU protein was significantly elevated in FED (p=0.006). RT-PCR analysis revealed that CLU mRNA was significantly increased (p= 0.008) in FED samples, but not in PBK samples. CLU had also a distinctive localization in FED samples with enhanced intracellular staining around the guttae and in the nuclei of endothelial cells.

Keywords: cornea: endothelium • degenerations/dystrophies • proteomics 
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