Clusterin protein diversity in the primate eye.
OBJECTIVE
The clusterin gene encodes a multi-functional protein that has been identified in different tissues, including a number of different eye tissues, primarily in the mouse and to a much lesser extent in humans. Clusterin has been implicated in a number of cellular processes such as lipid transport, membrane integrity, apoptosis, and neurodegeneration, all of which could be important to the biology of the eye. In the current communication, we provide data that confirms the expression of clusterin in a number of different human eye tissues and establishes the expression profile of this gene in monkey derived eye tissues. The issue that we sought to examine is whether a broad profile of clusterin expression in the eye is consistent in primates (monkey and human).


METHODS
The majority of our study was done using monkey eye tissues. Where possible, we have used human tissues in order to confirm published findings. Northern and western analysis was performed using tissues derived from monkey eyes. In situ hybridization and immunochemistry were carried out on human eye sections.


RESULTS
Clusterin mRNA is expressed in primate lens, cornea, limbus, sclera, orbital muscle, ciliary body, retina, RPE/choroid, and RPE cells in culture. Western analysis revealed that two major groups of clusterin exist in the eye, a high molecular weight group (>100 kDa) and a second group consisting of at least five clusterin species that are all approximately 80 kDa. Analysis of conditioned media from RPE cells cultured on permeable supports suggests that different forms of clusterin display alternative patterns of secretion.


CONCLUSIONS
Clusterin is expressed in a broad range of eye tissues in both human and monkey, suggesting that this is a characteristic feature in primates. We demonstrate for the first time that a diverse number of clusterin isoforms were observed in monkey eye tissues by western analysis. Meanwhile, the molecular size of clusterin mRNA detected in the array of tissues are identical in size, suggesting that the nature of the diversity in clusterin forms is due to post-translational modifications. In addition, new insights were made in defining clusterin expression in ciliary body, cornea, and the retinal pigment epithelium.
Wong
P
P
Departments of Biological Sciences, Ophthalmology, and Medical Genetics, University of Alberta, Edmonton, Alberta, Canada. pwwong@gpu.srv.ualberta.ca
Pfeffer
B A
BA
Bernstein
S L
SL
Chambers
M L
ML
Chader
G J
GJ
Zakeri
Z F
ZF
Wu
Y Q
YQ
Wilson
M R
MR
Becerra
S P
SP
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D051152:Clusterin; D003169:Complement Inactivator Proteins / Q000096:biosynthesis / Q000235:genetics*; D005123:Eye / Q000378:metabolism*; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D008253:Macaca mulatta; D018832:Molecular Chaperones; D008970:Molecular Weight; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*; D014018:Tissue Distribution
C495242:CLU protein, human; D051152:Clusterin; D003169:Complement Inactivator Proteins; D005136:Eye Proteins; D006023:Glycoproteins; D018832:Molecular Chaperones; D020033:Protein Isoforms; D012333:RNA, Messenger