retinol-acetate and Retinal-Degeneration

The retinoid visual cycle is an ocular retinoid metabolism specifically dedicated to support vertebrate vision. The visual cycle serves not only to generate light-sensitive visual chromophore 11-cis-retinal, but also to clear toxic byproducts of normal visual cycle (i.e. all-trans-retinal and its condensation products) from the retina, ensuring both the visual function and the retinal health. Unfortunately, various conditions including genetic predisposition, environment and aging may attribute to a functional decline of the all-trans-retinal clearance. To combat all-trans-retinal mediated retinal degeneration, we sought to slow down the retinoid influx from the RPE by inhibiting the visual cycle with a small molecule. The present study describes identification of CU239, a novel non-retinoid inhibitor of RPE65, a key enzyme in the visual cycle. Our data demonstrated that CU239 selectively inhibited isomerase activity of RPE65, with IC

Topics: Animals; cis-trans-Isomerases; Diterpenes; Enzyme Inhibitors; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Retinal Degeneration; Retinyl Esters; Vision, Ocular; Vitamin A

Exposure to bright light can cause visual dysfunction and retinal photoreceptor damage in humans and experimental animals, but the mechanism(s) remain unclear. We investigated whether the retinoid cycle (i.e. the series of biochemical reactions required for vision through continuous generation of 11-cis-retinal and clearance of all-trans-retinal, respectively) might be involved. Previously, we reported that mice lacking two enzymes responsible for clearing all-trans-retinal, namely photoreceptor-specific ABCA4 (ATP-binding cassette transporter 4) and RDH8 (retinol dehydrogenase 8), manifested retinal abnormalities exacerbated by light and associated with accumulation of diretinoid-pyridinium-ethanolamine (A2E), a condensation product of all-trans-retinal and a surrogate marker for toxic retinoids. Now we show that these mice develop an acute, light-induced retinopathy. However, cross-breeding these animals with lecithin:retinol acyltransferase knock-out mice lacking retinoids within the eye produced progeny that did not exhibit such light-induced retinopathy until gavaged with the artificial chromophore, 9-cis-retinal. No significant ocular accumulation of A2E occurred under these conditions. These results indicate that this acute light-induced retinopathy requires the presence of free all-trans-retinal and not, as generally believed, A2E or other retinoid condensation products. Evidence is presented that the mechanism of toxicity may include plasma membrane permeability and mitochondrial poisoning that lead to caspase activation and mitochondria-associated cell death. These findings further understanding of the mechanisms involved in light-induced retinal degeneration.

Topics: Acute Disease; Aging; Alcohol Oxidoreductases; Animals; Apoptosis; ATP-Binding Cassette Transporters; bcl-2-Associated X Protein; Caspases; Cell Line; Cell Survival; Chromatography, High Pressure Liquid; Chromatography, Liquid; Diterpenes; Ethanolamine; Humans; Light; Mass Spectrometry; Mice; Oxidation-Reduction; Rats; Retina; Retinal Degeneration; Retinal Diseases; Retinaldehyde; Retinyl Esters; Rhodopsin; Vitamin A

Mice lacking retinal pigment epithelium-specific 65-kDa protein (RPE65) develop retinopathy and blindness resembling Leber congenital amaurosis. Effects of 9-cis-retinyl acetate (9-cis-R-Ac) on visual function and retinopathy progression were tested in Rpe65(-/-) mice.. Young C57Bl/6 mice were given 9-cis-R-Ac in each of four different oil-based vehicle solutions by gastric gavage to identify the vehicle most suitable for drug delivery by measuring retinoid levels in plasma. Then doses of 9-cis-R-Ac ranging from 1 to 100 mg/kg were administered to 5- to 12-week-old Rpe65(-/-) mice by different treatment regimens, including single doses and either intermittent or daily doses for various periods up to 8 weeks. Retinoid effects on visual function were evaluated by electroretinography, retinoid analyses, histologic methods, and vision-dependent behavioral testing.. Soybean oil vehicle provided the highest 9-cis-R-Ac metabolite levels in plasma. Single doses of 9-cis-R-Ac (6.25-50 mg/kg) provided significant dose-dependent improvement in electroretinographic responses. Well-tolerated daily doses (1-12.5 mg/kg) for 2 weeks induced remarkable improvement of retinal function. Significant dose-dependent improvement of electroretinographic responses was observed 6 days after administration of 9-cis-R-Ac daily for 3 days at 1 to 12.5 mg/kg. Mice given either daily or intermittent 9-cis-R-Ac treatment at 1 and 4 mg/kg and evaluated 8 weeks later displayed dose-dependent improvement of retinal function and morphology, whereas retinal function deteriorated in control animals. Treated mice also performed better than control animals in vision-dependent behavioral tests.. Treatment with 9-cis-R-Ac improves visual function and preserves retinal morphology in Rpe65(-/-) mice.

Topics: Animals; Behavior, Animal; Blindness; Carrier Proteins; cis-trans-Isomerases; Diterpenes; Electroretinography; Eye Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Pharmaceutical Vehicles; Photic Stimulation; Prodrugs; Retina; Retinal Degeneration; Retinoids; Retinyl Esters; Soybean Oil; Vitamin A

Lipid hydroperoxides (LHP) have been synthesized and purified from linoleic, linolenic, arachidonic and docosahexaenoic acids, using soybean lipoxygenase and oxygen. Intravitreal injections into the eyes of mature, albino rabbits produced an early and then progressive decrease in the amplitude of a-, b- and c-waves of the ERG. Depending upon the amount and activity of the LHP preparation, ERG's were markedly decreased in amplitude (greater than 50%) within 4 days following the injection and by 12 days, the activity from peroxide treated eyes was essentially nonrecordable. In preliminary studies, these effects were less pronounced in adult pigmented rabbits of similar age, however, a younger pigmented rabbit was only slightly less susceptible to damage than the albino animals. In other experiments, peroxidized native phospholipids, malonaldehyde, hydrogen peroxide and sodium iodate were also shown to be cytotoxic, but not all were as toxic as the LHP. In contrast, retinol, vitamin A acetate and retinoic acid had no effect upon ERG activity, nor did the parent fatty-acid compounds or the borate buffer in which they were injected. These studies confirm previous reports where indirect production of lipid peroxides caused retinal degeneration. The present report extends these observations to demonstrate that when the retina and RPE are exposed to a sample of purified LHP, retinal function is altered in an irreversible way. We also demonstrate that a metabolic by-product (malonaldehyde) is likewise cytotoxic. However, the mechanisms by which the parent LHP and/or metabolites might act could be quite different. This new animal model should prove useful in evaluating further the ultrastructural changes which are observed during peroxidative damage of the retina in vivo, as well as in evaluating the therapeutic approaches to these problems of retinal degeneration.

Topics: Animals; Arachidonic Acids; Diterpenes; Docosahexaenoic Acids; Electroretinography; Leukotrienes; Linolenic Acids; Lipid Peroxides; Rabbits; Retina; Retinal Degeneration; Retinyl Esters; Vitamin A