PURPOSE: Heme oxygenase (HO)-1 immunoreactivity (IR) was examined in normal untreated retina and in retinal explants after in vitro treatment with stress agents. METHODS: Enucleated eyes from young adult C3H mice were immediately fixed and cryosectioned and the retina sections processed for immunocytochemistry with antibodies against HO-1 and glial fibrillary acidic protein (GFAP). From other eyes retinas were isolated and maintained in organ culture, either untreated for 4 days maximum or for 21 hours during which the explants were treated the first 3 hours with selected doses of sodium arsenate or hydrogen peroxide. Thereafter, the explants were processed identically with the normal tissue. RESULTS: In the normal retina, HO-1 and GFAP IR was very low. The culturing itself resulted in an increase in both HO-1 and GFAP immunolabeling in Müller cells of explanted retinas. Both sodium arsenate and hydrogen peroxide further induced strong HO-1 IR in Müller cells but not in other retinal cells. In contrast to HO-1, GFAP staining in Müller cells was not altered as a result of treatment, either by sodium arsenate or hydrogen peroxide at any concentration used. CONCLUSIONS: The results show for the first time that HO-1 can be induced in the retina in vitro by conditions of oxidative stress and that enzyme expression is confined exclusively to Müller cells.
PURPOSE: Heme oxygenase (HO)-1 immunoreactivity (IR) was examined in normal untreated retina and in retinal explants after in vitro treatment with stress agents. METHODS: Enucleated eyes from young adult C3H mice were immediately fixed and cryosectioned and the retina sections processed for immunocytochemistry with antibodies against HO-1 and glial fibrillary acidic protein (GFAP). From other eyes retinas were isolated and maintained in organ culture, either untreated for 4 days maximum or for 21 hours during which the explants were treated the first 3 hours with selected doses of sodium arsenate or hydrogen peroxide. Thereafter, the explants were processed identically with the normal tissue. RESULTS: In the normal retina, HO-1 and GFAP IR was very low. The culturing itself resulted in an increase in both HO-1 and GFAP immunolabeling in Müller cells of explanted retinas. Both sodium arsenate and hydrogen peroxide further induced strong HO-1 IR in Müller cells but not in other retinal cells. In contrast to HO-1, GFAP staining in Müller cells was not altered as a result of treatment, either by sodium arsenate or hydrogen peroxide at any concentration used. CONCLUSIONS: The results show for the first time that HO-1 can be induced in the retina in vitro by conditions of oxidative stress and that enzyme expression is confined exclusively to Müller cells.
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