Localization of corneal superoxide dismutase by biochemical and histocytochemical techniques

Redmond, T. Michael; Duke, E. J.; Coles, William H.; Simson, J. A. V.; Crouch, Rosalie K.

doi:10.1016/0014-4835(84)90192-1

Cited by 35 publications

(12 citation statements)

References 21 publications

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“…Indeed SIRC, being basically fibroblasts (keratocytes), possess a low (or absent) expression of some epithelial enzymes suitable to inhibit the ROS attack [48,55,56].…”

Section: Discussionmentioning

confidence: 99%

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Ciuffi

Pisanello

Pagliai

et al. 2003

Journal of Photochemistry and Photobiology B: Biology

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Several corneal pathologies are characterized by the presence of reactive oxygen species (ROS); therefore, we evaluated the protection afforded by pirenoxine and melatonin to corneal cell culture and whole rabbit cornea from ultraviolet exposure and other oxidant systems.Rabbit cornea cell (SIRC) plates and whole corneas were exposed to UV-B (80 or 800 mJ/cm 2 ) or incubated with fMLPstimulated autologous macrophages, in the presence or absence of pirenoxine or melatonin (10 À5 M). The protective activity of compounds was assessed by measuring superoxide anion formation, inhibition of oxidation and mitochondrial viability. Moreover the ex vivo protective effect of pirenoxine and melatonin was verified in the whole cornea submitted to UV-B exposure in vitro.Our experimental data demonstrate that pirenoxine and melatonin were able to inhibit the superoxide formation and oxidative effect in cell culture and whole rabbit corneas submitted to UV-B exposure or to incubation with fMLP-stimulated autologous macrophages.Mitochondrial viability was restored in epithelial cells of rabbit cornea but not in SIRCs. Moreover, both compounds are also able to increase ex vivo epithelial corneal cell defences against the in vitro UV-B induced lipid peroxidation.

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“…Indeed SIRC, being basically fibroblasts (keratocytes), possess a low (or absent) expression of some epithelial enzymes suitable to inhibit the ROS attack [48,55,56].…”

Section: Discussionmentioning

confidence: 99%

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Ciuffi

Pisanello

Pagliai

et al. 2003

Journal of Photochemistry and Photobiology B: Biology

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“…Likewise, some bacteria contain inducible proteins that have structural domains for binding both iron and DNA and are thought to confer resistance to peroxide damage during periods of oxidative stress (46). Transparent ocular tissues such as those of the cornea and lens are known to contain high concentrations of a number of components known to diminish damage from AOS, including superoxide dismutase (47,48). Numerous studies have demonstrated that iron greatly exacerbates AOS damage to DNA (49 -51).…”

Section: Discussionmentioning

confidence: 99%

Ferritin Is a Developmentally Regulated Nuclear Protein of Avian Corneal Epithelial Cells

Cai

Birk

Linsenmayer

1997

Journal of Biological Chemistry

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Previously, we generated monoclonal antibodies against chicken corneal cells (Zak, N. B., and Linsenmayer, T. F. (1983) Dev. Biol. 99, 373). We have now observed that one group of these antibodies reacts with a developmentally regulated component of corneal epithelial cell nuclei. This component is the heavy chain of ferritin, as determined by analyses of immunoisolated cDNA clones and immunoblotting of the protein. Immunoblotting also suggests that the nuclear ferritin may be in a supramolecular form that is similar to the ironbinding ferritin complex found in the cytoplasm of many cells. In vitro cultures and transfection studies show that the nuclear localization depends predominantly on cell type but can be altered by the in vitro environment. The appearance of nuclear ferritin is at least partially under translational regulation, as is known to be true for the cytoplasmic form of the molecule. The tissue and developmental distributions of the mRNA for the molecule are much more extensive than the protein itself, and the removal of iron from cultures of corneal epithelial cells with the iron chelator deferoxamine prevents the appearance of nuclear ferritin. At present the functional role(s) of nuclear ferritin remain unknown, but previous studies on cytoplasmic ferritin raise the possibility that it prevents damage due to free radical generation ("oxidative stress") by sequestering iron. Although it remains to be tested whether nuclear ferritin prevents oxidative damage, we find this an attractive possibility. Since the corneal epithelium is transparent and is constantly exposed to free radical-generating UV light, it is possible that the cells of this tissue have evolved a specialized mechanism to prevent oxidative damage to their nuclear components.Nuclei contain a myriad of different proteins as well as nucleic acids. Nuclear proteins range from general ones, such as the histones that are universally present in cells and determine the structure of chromatin, to the DNA-binding regulatory proteins, some of which are cell-specific and developmentally regulated. In the present investigation, we provide evidence that within the corneal epithelium of embryonic chicks, ferritin is also a developmentally regulated nuclear protein. In other cell types that have been investigated, the molecule is cytoplasmic and is thought to function as a protective antioxidant by sequestering iron (1, 2) (see "Discussion").In earlier work we sought to identify developmentally regulated components of the embryonic avian cornea by generating hybridomas against corneal cell suspensions (3). By immunofluorescence, one group of antibodies (including antibody 6D11) reacted with a corneal epithelial antigen but had little or no detectable reactivity with other tissues in the embryo. During development, this antigen first became detectable by immunofluorescence in 12-day corneas, the time when the epithelium begins to stratify (4). We also found it to arise de novo in cultured corneas, and it has been used as a marker for corneal epithe...

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“…Additionally, differences have been shown to exist within the cornea. For example, superoxide dismutase, catalase and glutathione peroxidase are present in the corneal epithelium and endothelium but absent or minimal in the stroma (2,3,13,57,72). Furthermore, ferritin, which protects against UV-induced, free-radical nuclear damage, is present in only the corneal epithelium (9).…”

Section: Changes In In Vivo CM Through Focusing (Ie Light Scattering)mentioning

confidence: 99%

“…Scavengers of reactive oxygen species are present in the eye and tears (2,3,9,12,13,57,62,64,65,72). They can be grouped into nutrients (eg, vitamins A, E, C), antioxidant enzymes (eg, superoxide dismutase, catalase, glutathione peroxidase, metallothionein), several low molecular weight substances (eg, uric acid, GSH, tyrosine), and pigments (eg, ferritin, xanthophylls, melanin, tryptophan oxidation products) (9,64).…”

Section: Changes In In Vivo CM Through Focusing (Ie Light Scattering)mentioning

confidence: 99%

Pathology of Ocular Irritation with Bleaching Agents in the Rabbit Low-Volume Eye Test

et al. 2001

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Despite differences in the processes leading to tissue damage, the ocular irritation response to various surfactants, two concentration s of an acid and an alkali, and an acetone, alcohol, aromatic amine, and aldehyde has been shown to depend on the extent of initial injury. The purpose of this study was to assess the extent to which this fundamenta l relationship exists for bleaching agents in the rabbit low-volume eye test. Ten l l of sodium perborate monohydrat e (NaBO 3 ), sodium hypochlorite (NaOCl), 10% hydrogen peroxide (H 2 O 2 ), and 15% H 2 O 2 was applied directly to the cornea of the right eye of each rabbit. Macroscopic assessments for irritation were made 3 hours after dosing and periodically until 35 days. Light microscopic examinations were conducted on tissues obtained at 3 hr and on 1, 3, and 35 days. In vivo confocal microscopy (CM) and measurements of dead corneal epithelial cells and keratocytes at 3 hours and 1 day were used to characterize quantitatively initial corneal injury, while in vivo CM performed at 3 hours and 1, 3, 7, 14, and 35 days was used to characterize quantitatively the corneal changes over time. The changes with NaBO 3 and NaOCl were consistent with mild irritancy. For both, corneal injury was limited to the epithelium and super cial stroma. The changes with 10% H 2 O 2 and 15% H 2 O 2 were consistent with severe irritation. Both concentrations affected the epithelium and deep stroma, with 15% H 2 O 2 also at times affecting the endothelium. However, unlike other irritants previously studied, with 10% H 2 O 2 and 15% H 2 O 2 there was an incongruit y between the extent of epithelial and stromal injury, with stromal injury being more extensive than epithelial injury. A similar, although less dramatic, effect was observed with NaBO 3 . Additionally, there was still signi cant keratocyte loss at 35 days with 10% H 2 O 2 and 15% H 2 O 2 even though the eyes at times were considered to be macroscopicall y normal. These observations highlight the need to include both epithelial and stromal component s in an ex vivo or in vitro alternative assay. In conclusion, these results continue to support and extend our hypothesis that ocular irritation is principally de ned by the extent of initial injury despite clear differences in the means by which irritants cause tissue damage. Importantly, we have identi ed unique differences in the ocular injury and responses occurring with bleaching agents that are important to consider in the development and validation of alternative ocular irritation tests to characterize a broad range of materials differing in type and irritancy.

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Localization of corneal superoxide dismutase by biochemical and histocytochemical techniques

Cited by 35 publications

References 21 publications

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Ferritin Is a Developmentally Regulated Nuclear Protein of Avian Corneal Epithelial Cells

Pathology of Ocular Irritation with Bleaching Agents in the Rabbit Low-Volume Eye Test

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