Immunolocalization of ubiquitin and related enzymes in human retina and retinal pigment epithelium.
OBJECTIVE
To examine the localization of ubiquitin (Ub) and related enzymes in human retina with emphasis on the retinal pigment epithelium (RPE)-Bruch's membrane complex.


METHODS
Thirty human eyes enucleated for various disease processes were examined. Immunohistochemistry was performed on paraffin sections using antibodies against Ub, Ub-conjugating enzyme (E2), Ub carboxyl-terminal hydrolase (PGP 9.5), and, for comparison, arrestin (Arr). Immunoreactivity (IR) was tested using the avidin-biotin method.


RESULTS
Ub was present throughout retina but was particularly prominent in ganglion cells and RPE. Most intriguing was the presence of Ub IR in age-related, sub-RPE deposits such as drusen and basal laminar deposits (BLD). RPE immunolabeling was more intense in older tissue, but otherwise no pattern of Ub IR could be linked to specific diseases. E2 IR colocalized with Ub, with one exception; E2 IR was not found in drusen or BLD. PGP 9.5 IR was intense in nerve fibers, ganglion cells, and the inner nuclear and plexiform layers. RPE staining was faint and patchy; sub-RPE deposits were not labeled. Arr IR was present in photoreceptors but not within or beneath RPE cells.


CONCLUSIONS
The ubiquitination process is important in human retina and particularly in ganglion cells. Ub-related processes are also active in RPE and may be involved in the degradation and disposal of proteins from these cells. The presence of Ub in sub-RPE deposits--without related Ub-processing enzymes--raises the possibility that certain proteins become ubiquitinated within RPE but that further degradation of the Ub-protein complexes does not occur.
Loeffler
K U
KU
Department of Ophthalmology, Freiburg University, Germany.
Mangini
N J
NJ
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019393:Arrestin / Q000378:metabolism; D005128:Eye Diseases / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008025:Ligases / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010857:Pigment Epithelium of Eye / Q000201:enzymology / Q000378:metabolism*; D012160:Retina / Q000378:metabolism* / Q000473:pathology; D013869:Thiolester Hydrolases / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase; D044763:Ubiquitin-Conjugating Enzymes; D014452:Ubiquitins / Q000378:metabolism*
D019393:Arrestin; D009419:Nerve Tissue Proteins; D014452:Ubiquitins; D044763:Ubiquitin-Conjugating Enzymes; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; D008025:Ligases