Identification and localization of retinal cystatin C.
OBJECTIVE
Cystatin C is a mammalian cysteine protease inhibitor, synthesized in various amounts by many kinds of cells and appearing in most body fluids. There are reports that it may be synthesized in the mammalian retina and that a cysteine protease inhibitor may influence the degradation of photoreceptor outer segment proteins. In the current study cystatin C was identified, quantitated, and localized in mouse, rat, and human retinas.


METHODS
Enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), DNA sequencing, Western blot analysis, and immunohistochemistry have been used on mouse, rat, and human retinas (pigment epithelium included).


RESULTS
Cystatin C is present in high concentrations in the normal adult rat retina, as it is throughout its postnatal development. Its concentration increases to a peak at the time when rat pups open their eyes and then remains at a high level. It is mainly localized to the pigment epithelium, but also to some few neurons of varying types in the inner retina. Cystatin C is similarly expressed in normal mouse and human retinas.


CONCLUSIONS
Cystatin C was identified and the localization described in the retinas of rat, mouse, and human using several techniques. Cystatin C is known to efficiently inactivate certain cysteine proteases. One of them, cathepsin S, is present in the retinal pigment epithelium and affects the proteolytic processing by cathepsin D of diurnally shed photoreceptor outer segments. Hypothetically, it appears possible that retinal cystatin C, given its localization to the pigment epithelium and its ability to inhibit cathepsin S, could be involved in the regulation of photoreceptor degradation.
Wassélius
J
J
Department of Ophthalmology, Lund University Hospital, S-221 84 Lund, Sweden. johan.wasselius@oft.lu.se
Håkansson
K
K
Johansson
K
K
Abrahamson
M
M
Ehinger
B
B
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015153:Blotting, Western; D055316:Cystatin C; D015891:Cystatins / Q000032:analysis*; D015853:Cysteine Proteinase Inhibitors / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D010857:Pigment Epithelium of Eye / Q000737:chemistry; D051381:Rats; D012160:Retina / Q000737:chemistry* / Q000254:growth & development; D020133:Reverse Transcriptase Polymerase Chain Reaction
C525030:CST3 protein, human; C525031:Cst3 protein, mouse; C525032:Cst3 protein, rat; D055316:Cystatin C; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors