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Observations on the retinal pigment epithelium and retinal macrophages in experimental retinal detachment.

After experimental retinal detachment in rabbits macrophages are a prominent feature in the subretinal space or within the retina. Two sources for these macrophages are identified. The retinal pigment epithelium (RPE) may undergo metaplasia and actively 'bud'; the evolving macrophage is then formed by a vitreal protrusion of the cytoplasm of an RPE cell which is 'nipped off' by lateral protrusions from adjacent cells. In addition, in regions of RPE proliferation, blood-borne cells were found in Bruch's membrane and among the mass of proliferated RPE cells, suggesting that blood-borne cells may pass from the choroidal circulation through Bruch's membrane and the RPE layer.

Animals

Defective phagocytosis of isolated rod outer segments by RCS rat retinal pigment epithelium in culture.

Retinal pigment epithelium cultured from normal rats phagocytizes large amounts of rod outer segment fragments isolated from normal rats and from RCS rats with inherited retinal degeneration. Cultured RCS rat pigment epithelium rarely ingests outer segment material, although the cells extend cellular processes around fragments of either type. Both normal and RCS pigment epithelium phagocytize polystyrene spheres. This demonstrates that RCS rat pigment epithelial cells contain a defect in the mechanism for phagocytizing outer segments.

Animals

[Pigmentation of feather primordium achieved by grafting of retinal pigment epithelium].

Small fragments of the retinal pigmented epithelium from White Leghorn, Rhode Island Red chick embryos, and from Quail, Starling and Red Partridge embryos, were grafted into the wing bud of White Leghorn chick embryos. Retinal pigmented epithelium of White Leghorn chick embryos did not give rise to pigmentation of feather primordia in the hosts. In contrast, 60% of grafts from Rhode Island Red, Quail, Starling and Red Partridge, gave rise to feather primordia whose pigmentation was identical to that of the donors. Pigmentation depends on the fact that some graft retinal melanocytes diffuse in the wing bud of the host and reach the feather primordia. This was shown by the Feulgen technique in embryos grafted with Quail retinal pigmented epithelium, taking advantage of the fact that Quail cells possess DNA rich nucleoli. The local factors which allow the migration of the melanocytes are probably inactive on the retinal melanocytes of the White Leghorn chick embryos, whose genotype does not contain the "feather pigmentation" character.

Animals

Retinal vascular changes in congenital hypertrophy of the retinal pigment epithelium.

The overlying retinal blood vessels were abnormal in five cases of congenital hypertrophy of the retinal pigment epithelium. This illustrated the well-recognized association between outer retinal degeneration and obliteration of the overlying retinal vasculature. The proposed pathophysiological mechanisms, however, seem inadequate to explain completely the morphological changes of the retinal blood vessels in the presence of atrophy of the outer retina.

Adult

Microperoxisomes in retinal pigment epithelium.

Microperoxisomes were found to be abundant in the retinal pigment epithelium of the human, rhesus monkey, mice, rats, domestic fowl, and frog by ultrastructural histochemistry. They were rare in other cells of the retina and choroid. These organelles had a granular matrix, ranged in diameter from 0.15 mum to 0.30 mum, and were bound by a single tripartite membrane which often maintained slender connections with the smooth endoplasmic reticulum and other microperoxisomes. They exhibited a positive reaction (electron opaque product) following incubation in diaminobenzidine and H2O2 for the demonstration of the peroxidatic activity of catalase (Novikoff et al., J. Histochem. Cytochem. 20: 1006, 1972). The reaction was inhibited by: (1) aminotriazole; (2) dichlorophenol-indophenol; (3) preheating at 95 degrees C.; or (4) elimination of H2O2. Microperoxisomes, like the well-known peroxisomes (microbodies) of liver cells have been inplicated in various aspects of lipid metabolism and the detoxification of H2O2. We demonstrated for the first time that microperoxisomes respond to drug-induced changes in lipid metabolism, as previously shown for peroxisomes. Nafenopin is a recently utilized drug which greatly decreases serum lipids, increases hepatic catalase activity, and induces an increased size and number of hepatic peroxisomes. Black, beige, albino, and obese mutant mice of the C57BL/6J strain treated with nafenopin for several weeks showed a two- to threefold increase in the number of microperoxisomes in the retinal pigment epithelium. Microperoxisomes of the retinal pigment epithelium may be involved in the transport, storage, and rapid turnover of lipids associated with the maintenance of photoreceptor outer segment disc membranes.

Adult

Primary retinal degeneration: evidence of normal phagocytosis in the retinal pigment epithelium.

In rats with primary retinal degeneration, lens extraction combined with total retinal detachment provided a model for injection of a tracer of colloidal carbon into the subretinal space. Electron microscopy and acid phosphatase cytochemistry were subsequently used to analyze the ingestion of tracer by the retinal pigment epithelium. It was found that the attachment, ingestion, and digestion phases of the phagocytic process were apparently preserved. From this evidence it is suggested that there is no lack of phagocytic power in the retinal pigment epithelium of affected rat strains.

Acid Phosphatase

Hemorrhagic detachment of the peripheral retinal pigment epithelium.

Six patients had hemorrhagic detachment of the peripheral retinal pigment epithelium. Abnormal neovascular nets in older individuals, under the retinal pigment epithelium and originating from the choriocapillaris, may well be the cause of such hemorrhagic detachments. This lesion must be differentiated from choroidal tumors, choroidal detachment, and uveal effusion.

Adult

Accumulation of taurine by cultured retinal pigment epithelium of the rat.

Cultured rat retinal pigment epithelium actively accumulates taurine against a concentration gradient; an intracellular concentration of 12 mM is maintained at an extracellular taurine concentration of 0.12 mM. Taurine accumulation conforms to Michaelis-Menten kinetics for extracellular taurine concentrations between 0.006 and 0.05 mM; within that range the Km is 16 muM and the Vmax is 2.3 nmol./hr./mul of cells. Above an extracellular concentration of 0.5 mM the kinetics are complex. Accumulation is inhibited by reduced temperature and iodoacetate as well as by dinitrophenol in the absence of glucose. Ouabain, reduced extracellular sodium, and removal of extracellular potassium and calcium also inhibit taurine accumulation. The cells also release previously accumulated taurine at half-time of 1 to 3 hours. The rapid accumulation and release of taurine by cultured retinal pigment epithelial cells is consistent with the idea that this tissue is a site for the transport of taurine in vivo.

Animals

Phagocytosis in the retinal pigment epithelium of the RCS rat.

The retinal pigment epithelium of RCS rats, previously thought not to phagocytize photoreceptor outer segments, exhibited a peak of phagocytosis in vivo when animals were kept under conditions of cyclic lighting (12 hours of darkness and 12 hours of light). The peak occurred at 1 hour after the onset of light, with maximum and minimum levels of phagocytosis averaging about 5 percent of that found in the pigment epithelium of Osborn-Mendel rats used as a control. Eyecups that were obtained from Osborn-Mendel rats and maintained for up to 3 hours in organ culture demonstrated levels of phagocytosis that were sevenfold greater than those of unincubated controls. Likewise a tenfold increase occurred in incubated as opposed to unicubated RCS eyes, raising the possibility that phagocytosis could be experimentally stimulated in vivo.

Animals

Culture of rat retinal pigment epithelium.

A method of preparing monolayer cultures of retinal pigment epithelium from normal pigmented neonatal rats is described. Critical features include incubating the eyes in balanced salt solution and treating with trypsin before dissecting the eyes. The tissue also has been culured from RCS rats with inherited retinal degeneration. Since the pigment epithelium has been shown to be the primary site of action of the gene for retinal dystrophy in the RCS rat, the method should be usefull in studying the defect(s) associated with this mutation.

Animals

Patterned dystrophies of the retinal pigment epithelium.

Three varieties of patterned dystrophies of the retinal pigment epithelium have been described in the literature: They are reticular dystrophy, macroreticular dystrophy, and butterfly-shaped pigment dystrophy of the fovea (macula). They are all inherited and are bilateral and symmetric. Results of retinal-function tests in all three varieties were essentially normal, except for the electro-oculogram in the butterfly-shaped pigment dystrophy. We have observed within a single family one member manifesting butterfly-shaped pigment dystrophy, one member with macroreticular dystrophy, and one member with probable reticular dystrophy. Electrophysiologic studies of the retina in these patients were in accord with those already reported, except for supernormal scotopic responses in the latter two patients. This family demonstrates the linkage necessary to tie together a number of entities heretofore considered as separate.

Adult

Some observations in the fine structure of age changes of the mouse retinal pigment epithelium.

Age changes of the fine structure in the CF-1 mouse retinal pigment epithelium were studied from 1--24 months old. The change most characteristic of the senescence of the retinal pigment epithelium was found to be the appearance of wavy undulating 12nm wide filaments having a periodicity of about 80nm after the 12th month. In addition, the retinal pigment epithelial cells of the aging eye were found to have a marked accumulation of lysosomal dense bodies measuring about 0.3 to 1.2 micrometer in diameter especially in the apical portion of the cytoplasm. Extended and enlarged basal cytoplasmic infoldings of the retinal pigment epithelial cells were another prominent age change in the old mouse. In these basal infoldings densely fixed amorphous materials by tannic acid and very fine filaments are recognized. Age-related changes of the retinal pigment epithelial cell of the CF-1 mouse may take place as the degeneration of cytoplasmic materials, the appearance of bundles consisting of parallel undulating filaments, and the extending process of basal infolding from the choroid side.

Aging

Natural history of serous detachments of the retinal pigment epithelium.

Fifty eyes of 40 patients with serous (avascular) detachments of the retinal pigment epithelium were followed up for an average of 22 months. Of the 50 eyes, 14 (28%) developed bleeding beneath the retinal pigment epithelium and retina or subretinal neovascularization detected on fluorescein angiography. These changes did not occur in eyes of patients younger than age 56 years at diagnosis or in detachments less than 1 disk diameter in size, whereas 35.9% (14) of eyes in patients older than age 56 years developed vascular complications. Vascular complications were uncommon when the initial detachment did not involve the fovea. Visual acuity declined in only one (9.1%) of eyes in patients younger than age 56 years but in 20 (51.3%) of eyes in patients age 56 years or greater.

Adult

On the retinal pigment epithelium of the barn owl (Tyto alba).

The fine structure of the retinal pigment epithelium (RPE) of the barn owl (Tyto alba) is described. In general it resembles that of the retinal pigment epithelium of other vertebrate animals and a high content of myeloid bodies, lipid droplets, electron dense cytosomes, and especially phagosomes is observed. The latter show different structural specializations which include membranes, tubules, filaments, granules, vesicles, and crystalloid patterns. The central part of the retina often contains a peculiar inclusion body which is separated from the cytoplasm by one or two distinct membranes and is composed of a regularly arranged tubular network which forms a three-dimensional honeycomb structure. The tubules in part are in continuation with randomly oriented and densly packed tubules and stacks of discs consisting of paired lamellae. Similar formations within secretory cells of a gland (dendritic organ) and the taste buds of catfish are discussed.

Animals

Localization of lysosomal enzymes in retinal pigment epithelium of rats with inherited retinal dystrophy.

Four acid hydrolase activities are demonstrable by light microscopy in pigment epithelial cell lysosomes of rats (Royal College of Surgeons--RCS) with inherited retinal dystrophy and in control (Fischer) rats. The enzymes include acid phosphatase, aryl sulfatase, N-acetyl-beta-glucosaminidase, and esterase activities. No marked differences are observed in distribution or staining intensity of lysosomes in the two strains of rat. Acid hydrolase activities are not localized in sites other than lysosomes. Acid phosphatase and aryl sulfatase activities are also demonstrable by electron microscopy. In both strains, acid phosphatase reaction product is localized to various forms of lysosomes in pigment epithelial cells. A diffuse precipitate, considered to be nonspecific in origin, is seen in the cytoplasm, apical processes, outer segments (control), and outer segment debris (RCS). The precipitate is probably due to adsorption of lead from the incubation medium or of lead phosphate that diffuses from heavy accumulations in nearby lysosomes. Aryl sulfatase reaction product, in contrast to acid phosphatase, is localized to far fewer lysosomes and there is virtually no nonspecific precipitate. The findings indicate that lysosomes of RCS pigment epithelial cells possess several cytochemically demonstrable acid hydrolase activities. There is no evidence for the localization of acid phosphatase (or aryl sulfatase) activities in sites other than lysosomes.

Acid Phosphatase

Effects of colchicine on phagosome-lysosome interaction in retinal pigment epithelium. I. In vivo observations in albino rats.

The retinal pigment epithelium (RPE) from young albino rats was examined under light and electron microscope 2 1/2 to 16 h after intravenous injections of colchicine (0.5 mg per 100 gm body weight). One of the primary effects of colchicine treatment on the RPE is the appearance of extensive acPase-positive dense body aggregates in the Golgi area. The size of the aggregates and the disappearance of microtubules appear to be related to the length of time allowed for colchicine to react. Acid phosphatase (acPase) activity is localized on the dense body aggregates and is particularly evident at a light microscopic level, where it can be observed as darkly stained clumps. Dense bodies are only rarely seen in the immediate proximity of acPase-positive phagosomes in colchicine-treated animals. The results of these experiments suggest the existence of an intracellular transport mechanism for acPase-carrying dense bodies. This intracellular transport is inhibited by colchicine, being probably related to a block of microtubular assembly.

Animals