Identification and localization of retinal cystatin C

PURPOSE. Cystatin C is a mammalian cysteine protease inhibitor, synthesized in various amounts by many kinds of cells and appearing in most body fluid s. There are reports that it may be synthesized in the mammalian retina and that a cysteine protease inhibitor may influence the degradation of photor eceptor outer segment proteins. In the current study cystatin C was identif ied, quantitated, and localized in mouse, rat, and human retinas. METHODS. Enzyme-linked immunosorbent assay (ELISA), reverse transcription-p olymerase chain reaction (RT-PCR), DNA sequencing, Western blot analysis, a nd immunohistochemistry have been used on mouse, rat, and human retinas (pi gment epithelium included). RESULTS. Cystatin C is present in high concentrations in the normal adult r at 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 rem ains 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 i s similarly expressed in normal mouse and human retinas. CONCLUSIONS. Cystatin C was identified and the localization described in th e retinas of rat, mouse, and human using several techniques. Cystatin C is known to efficiently inactivate certain cysteine proteases. One of them, ca thepsin S, is present in the retinal pigment epithelium and affects the pro teolytic processing by cathepsin D of diurnally shed photoreceptor outer se gments. Hypothetically, it appears possible that retinal cystatin C, given its localization to the pigment epithelium and its ability to inhibit cathe psin S, could be involved in the regulation of photoreceptor degradation.