Clusterin deficiency in eyes with pseudoexfoliation syndrome may be implicated in the aggregation and deposition of pseudoexfoliative material.
OBJECTIVE
To determine the role of the extracellular chaperone clusterin in the pathophysiology of pseudoexfoliation (PEX) syndrome/glaucoma, which is characterized by the stable deposition of abnormal extracellular fibrillar material in anterior segment tissues.


METHODS
Real-time PCR, in situ hybridization, and immunohistochemistry were applied to analyze the mRNA and protein expression of clusterin in PEX eyes of patients without and with glaucoma and to compare them with eyes of patients with primary open-angle glaucoma and angle-closure glaucoma and with normal control eyes. Aqueous levels of clusterin were determined by Western blot analysis. Real-time PCR and Western blot analysis were used to study the effect of TGF-beta1, which is significantly increased in the aqueous humor of PEX eyes, on clusterin expression by nonpigmented ciliary epithelial cells in vitro.


RESULTS
Clusterin mRNA was ubiquitously expressed in most ocular cells and tissues, particularly in the epithelium of ciliary processes, whereas the protein was mostly located to extracellular structures, such as ocular basement membranes and stromal fibers. Real-time PCR and in situ hybridization displayed significant downregulation of clusterin mRNA in all anterior segment tissues of PEX eyes, irrespective of the presence or type of glaucoma, compared with normal and glaucomatous control eyes, whereas posterior segment tissues did not show any differential expression. A generally decreased immunoreactivity, but a prominent binding of clusterin to all PEX deposits, could be observed in ocular tissues of PEX eyes. Clusterin levels in aqueous humor were significantly reduced in eyes of patients with PEX syndrome compared with normal and glaucomatous control eyes. The expression of clusterin mRNA and protein in nonpigmented ciliary epithelial cells was significantly downregulated by TGF-beta1 in vitro.


CONCLUSIONS
Considering the known role of clusterin as a highly efficient extracellular chaperone, its deficiency in the anterior segment of PEX eyes may promote the stress-induced aggregation and stable deposition of the pathologic extracellular matrix product characteristic of PEX syndrome.
Zenkel
Matthias
M
Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany. matthias.zenkel@augen.imed.uni-erlangen.de
Kruse
Friedrich E
FE
Jünemann
Anselm G
AG
Naumann
Gottfried O H
GO
Schlötzer-Schrehardt
Ursula
U
D000368:Aged; D000369:Aged, 80 and over; D000869:Anterior Eye Segment / Q000378:metabolism; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002924:Ciliary Body / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051152:Clusterin / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D017889:Exfoliation Syndrome / Q000150:complications / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D015812:Glaucoma, Angle-Closure / Q000150:complications / Q000378:metabolism*; D005902:Glaucoma, Open-Angle / Q000150:complications / Q000378:metabolism*; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
C495242:CLU protein, human; D051152:Clusterin; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1