Comparative histochemical and immunohistochemical study on xanthine oxidoreductase/xanthine oxidase in mammalian corneal epithelium

We have previously found that xanthine oxidase (one form of xanthine oxidoreductase that generates reactive oxygen species, such as superoxide radicals and hydrogen peroxide) is present in corneal epithelium of norma; rabbit eye. It was suggested that the reactive oxygen species contribute to additional eye damage related to prolonged continuous contact tens wear and irradiation of the eye with UV-B light. To further explore the potential danger of xanthine oxidase as a source of reactive oxygen species, we have examined in the present paper whether xanthine oxidoreductase and xanthine oxidase are present in corneal. epithelium of other mammalian species, employing immunohistochemical and enzyme histochemical methods. In corneal epithelium of normal eyes of ox, pig, guinea-pig, and rat xanthine oxidoreductase activity was detected by the tetrazolium salt reduction method and xanthine oxidase activity was localized by a method based on cerium ions capturing hydrogen peroxide. For the immunohistochemical demonstration of the enzymes, rabbit anti-bovine xanthine oxidase antibody, rabbit anti-human xanthine oxidase antibody and monoclonal mouse anti-human xanthine oxidase/xanthine dehydrogenase/aldehyde oxidase antibody were used. The immunohistochemical and enzyme histochemical results show that xanthine oxidoreductase and xanthine oxidase are present both as proteins and as active enzymes in the corneal epithelium of all animals studied. It is hypothesized that under various pathological states, xanthine oxidase-generated reactive oxygen species might contribute to eye damage. (C) 2003 Elsevier GmbH. All rights reserved.