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