Retinol and retinyl esters in pigment epithelium of rats with inherited retinal degeneration

Summary A comparative study of the retinol and retinyl ester concentrations was performed in the retinal pigment epithelium of the normal and affected rats. Our findings indicate that in dystrophic rat retinol content increases, whereas the amount of retinyl esters is always lower than normal. An hypothesis can be made on the deficiency of enzymic activities which regulate retinol retinol and retinyl esters levels in the pigment epithelium.     

Competition between retinol and 3,4-didehydroretinol for esterification in crude pigment epithelial cell fractions

The membrane fraction of the retinal pigment epithelium (RPE) of the frog (Rana pipiens) catalyzed the esterification of tritiated retinol to retinyl esters. This esterification reaction was inhibited in the presence of 3,4-didehydroretinol.     

Competition between retinol and 3,4-didehydroretinol for esterification in crude pigment epithelial cell fractions

Summary The membrane fraction of the retinal pigment epithelium (RPE) of the frog (Rana pipiens) catalyzed the esterification of tritiated retinol to retinyl esters. This esterification reaction was inhibited in the presence of 3,4-didehydroretinol.     

Comparison of retinal pigment epithelium cell preparations from the bovine eye

Retinal pigment epithelium (RPE) cells were collected from bovine eyes using a new method. The cells were harvested by vortexing the RPE and underlying choroid in 0.05 M citrate phosphate buffer, pH 5. RPE cells recovered by this method were compared to a standard method by microscopic examination of cell integrity, estimation of total protein, and assay of 11-cis and all-trans retinyl ester hydrolase (REH) activities. Results suggest that this method collects RPE cells of good integrity and with a significantly higher protein yield than the conventional method. Additionally, a much higher retinyl ester hydrolase activity was noted. Therefore we propose that this procedure offers a new and convenient method in the collection of RPE proteins for certain purposes such as enzyme purification.     

Comparison of retinal pigment epithelium cell preparations from the bovine eye

Summary Retinal pigment epithelium (RPE) cells were collected from, bovine eyes using a new method. The cells were harvested by vortexing the RPE and underlying choroid in 0.05 M citrate phosphate buffer, pH 5. RPE cells recovered by this method were compared to a standard method by microscopic examination of cell integrity, estimation of total protein, and assay of 11-cis and all-trans retinyl ester hydrolase (REH) activities. Results suggest that this method collects RPE cells of good integrity and with a significantly higher protein yield than the conventional method. Additionally, a much higher retinyl ester hydrolase activity was noted. Therefore we propose that this procedure offers a new and convenient method in the collection of RPE proteins for certain purposes such as enzyme purification.     

Multinucleated cells in the retinal pigment epithelium: A scanning electron microscopic study

Summary Multinucleated cells in the retinal pigment epithelium of the albino rat are shown with the scanning electron microscope beside normal mononucleated cells. We suggest that such elements are formed during foetal life, owing to the absence of any mitotic activity in adult rats.     

Visual pigment regeneration: occurrence in frog retina upside down upon the pigment epithelium

Visual pigment regeneration in the frog requires apposition between the retina and retinal pigment epithelium. However, regeneration occurs nearly as well with the retina upside down (i.e. with the inner limiting membrane facing the pigment epithelium) as right side up.     

Visual pigment regeneration: Occurrence in frog retina upside down upon the pigment epithelium

Summary Visual pigment regeneration in the frog requires apposition between the retina and retinal pigment epithelium. However, regeneration occurs nearly as well with the retina upside down (i.e. with the inner limiting membrane facing the pigment epithelium) as right side up.Acknowledgments. Supported in part by the Medical Research Service of the Veterans Administration and National Eye Institute Grant EY-01678.     

Carbonic anhydrase type II in regenerating retinal pigment epithelium. A histochemical study in the rabbit

Type II carbonic anhydrase (CAII) in the cytoplasm of the retinal pigment epithelium (RPE) may contribute to the transport of water and solutes across the RPE. The activity of this enzyme in RPE during its response to damage, e.g., during regeneration, is therefore of interest in understanding retinal disease. Immuno-histochemistry was used to compare CAII activity of normal RPE and RPE experimentally induced to regenerate. In normal rabbits, the RPE stained intensely with a peroxidase-linked antibody specific for human CAII. Regenerating RPE stained less intensely. Within the regenerating epithelium, staining appeared more intense in mature cells than in immature ones, suggesting that CAII activity gradually returns during RPE regeneration.     

Persistence of added retinoids in organ culture media during induction of mucous metaplasia and glandular morphogenesis in hamster cheek pouches

The retinoid concentration (determined colorimetrically) did not change significantly in retinyl acetate-supplemented (6 micrograms/ml) Eagle's Minimal Essential Medium containing 10% fetal calf serum when stored at -20 or 4 degrees C over 7 days. After the medium was incubated at 37 degrees C for 48 h, 37-49% of the retinoid remained, whether or not tissue (neonatal Syrian hamster cheek pouch) was present, and irrespective of explant age. The normal retinoid level in the tissue was approximately 0.25 micrograms per gram. Therefore, neonatal hamster cheek pouches, incubated in medium with the addition of 6 micrograms of retinyl acetate per ml of medium and undergoing mucous metaplasia and some mucous gland morphogenesis, were continually being exposed to retinoid levels which, though gradually decreasing, remained well above their normal physiological level.     

Plasma and hepatic antioxidant control and vitamin A nutritional status

Twenty-seven rats were divided into three groups and fed on diets containing 0.3, 6 or 60 RE (retinol equivalent) retinyl palmitate/g food. After 7 weeks, hepatic vitamin A uptake was found to be more efficient in vitamin A-deficient rats than in rats given adequate vitamin A. We showed that during the metabolic adaptation of the animals to the level of vitamin A in the diet, extensive modifications occur in the antioxidant defences of the organism. In parallel with the increase in the level of vitamin A, the decrease in the level of -tocopherol in the plasma can bring about a greater susceptibility of the lipoproteins to oxidative stress. Similarly, the decrease in the hepatic -tocopherol level and in glutathione peroxidase activity leads to the weakening of the liver's antioxidant defences.     

Autoradiographic evidence for binding of 3H-flunitrazepam (Rohypnol) to melanin granules in the cat eye

This autoradiographic study revealed after an i.,v. injection an accumulation of 3H-flunitrazepam in melanin granules of the pigment epithelium and of the choroid cells. It seems that after a short period, a large proportion of the strongly bound drug is released.     

Persistence of added retinoids in organ culture media during induction of mucous metaplasia and glandular morphogenesis in hamster cheek pouches

Summary The retinoid concentration (determined colorimetrically) did not change significantly in retinyl acetatesupplemented (6 g/ml) Eagle's Minimal Essential Medium containing 10% fetal calf serum when stored at –20 or 4°C over 7 days. After the medium was incubated at 37°C for 48 h, 37–49% of the retinoid remained, whether or not tissue (neonatal Syrian hamster cheek pouch) was present, and irrespective of explant age. The normal retinoid level in the tissue was approximately 0.25 g per gram. Therefore, neonatal hamster cheek pouches, incubated in medium with the addition of 6 g of retinyl acetate per ml of medium and undergoing mucous metaplasia and some mucous gland morphogenesis, were continually being exposed to retinoid levels which, though gradually decreasing, remained well above their normal physiological level.     

The major carotenoid pigments of six species of barnacle

Résumé L'astaxanthine (et ses esters) est le caroténoïde principal de 6 espèces de balanes, et la zeaxanthine est le principal xanthophylle neutre. Les différences de composition du pigment des espèces sont probablement liées aux proportions relatives de la matière animale et végétale dans ses aliments.     

Regulation of the retinal interphotoreceptor matrix Na by the retinal pigment epithelium during the light response

We examined the rabbit retinal pigment epithelium (RPE) for Na transport properties which would allow it to buffer undesirable changes in Na concentrations in the interphotoreceptor matrix (IPM) during light and dark cycles. The RPE is selectivity permeable to sodium. Open and short circuit transport studies with RPE indicate a circulating (choroid to retina and back) Na current which does not compromise the electrical integrity of the blood brain barrier but together with the Na permeselectivity is of sufficient magnitude to buffer both upwards and downwards movements of IPM [Na] during light or dark responses.     

Intercellular junctions of hyperplastic retinal pigment epithelium

In rats with retinopathies induced by excess fluorescent light or injections of urethane, the retinal pigment epithelium (RPE) undergoes focal hyperplasia. Neither intravascularly injected horseradish peroxidase or lanthanum nitrate penetrated the sensory retina at these hyperplastic sites. Electron microscopy revealed that this was due to the persistence of intact tight junctions among a single layer of hyperplastic cells facing the sensory retina. These junctions prevented intraocularly injected microperoxidase from passing as well. Cells within the hyperplastic foci were connected only by adherent junctions that presented no permeability barrier.     

Light dependent accumulation of macrophages at the photoreceptor-pigment epithelial interface in the retina of albino mice

Summary In the subretinal space of albino mice, macrophages appear from the time of eye opening and increase in number for 6 months; thereafter they decline with age. Dark rearing retards the accumulation of these cells, and exposure to constant light results in a rapid increase. Observations suggest that macrophages appear as a response to visual cell decay in albino mice and supplement the phagocytic activity of the pigment epithelium.We thank R.K. Hawkins for efficient management of the animal colony, H.G. Jansen for cooperation in the use of electron microscope and Ms Paula van Alphen for help in preparation of the illustrations.     

Photoreceptor damage following exposure to excess riboflavin

Flavins generate oxidants during metabolism and when exposed to light. Here we report that the photoreceptor layer of retinas from black-eyed rats is reduced in size by a dietary regime containing excess riboflavin. The effect of excess riboflavin was dose-dependent and was manifested by a decrease in photoreceptor length. This decrease was due in part to a reduction in the thickness of the outer nuclear layer, a structure formed from stacked photoreceptor nuclei. These changes were accompanied by an increase in photoreceptor outer segment autofluorescence following illumination at 328 nm, a wavelength that corresponds to the excitation maxima of oxidized lipopigments of the retinal pigment epithelium.     

Autoradiographic evidence for binding of3H-flunitrazepam (Rohypnol®) to melanin granules in the cat eye

Summary This autoradiographic study revealed after an i.v. injection an accumulation of³H-flunitrazepam in melanin granules of the pigment epithelium and of the choroid cells. It seems that after a short period, a large proportion of the strongly bound drug is released.Acknowledgments. I thank Dr E. Kovács and Mrs V. Beuck for help in treating the animals and Miss C. Böhler and Mr R. Krauer for excellent technical assistance.This drug, with a radiochemical purity of 99.2%, was prepared by Drs R. Schwob and J. Würsch, F. Hoffmann-La Roche & Co., Ltd, Basel, Switzerland.     

Light and temperature affect retinyl ester hydrolase activity and visual pigment composition

Summary Dim light, in combination with high water temperature, resulted in a significant increase in the retinyl ester hydrolase activity in the goldfish retina. This rise in enzyme activity may relate to a selective increase in the availability of retinal chromophores thereby favoring the formation of rhodopsin under these light and temperature conditions.