Radioautographic study of melanin synthesis in cells of the pigmented epithelium of the retina in adult tritons following surgical removal of the retina]

The results are given for the autoradiographic study of melanin synthesis in the pigment epithelium cells of eye retina in the adult newts by the incorporation of 3H-dioxyphenylalanine (3H-DOPA). The synthesis of melanin began 4--8 days after the operation in the peripheral parts of the eye and but it was not observed in the central region of the eye fundus even 25 days alter the operation. A suggestion is put forward to the effect of the existence of different mechanisms for the initiation of melanin synthesis in these subpopulations of pigment epithelium.     

The retinal pigment epithelium and photoreceptor cells light-and electron microscopic studies on monkey eyes

Summary In the perifoveal retina of the monkey, Cercopithecus aethiops, the melanin granules are accumulated in apical cytoplasmatic protrusions of the pigment epithelial cells, facing the end of the cones. The rods are inserted deeper into the pigment epithelium than the cones; they reach the bottom of the infoldings of the apical surface membrane of the pigment epithelial cells. No melanin granules or other inclusions are situated at the end of the rods. The outer extremity of the rods is considerably inclined and in sections often appears as groups of rod discs which are incompletely or completely separated from the main part of the outer segments. This separation is regarded as an artifact caused by the inclination of the rods, and it is therefore not considered to represent phagocytosis of the outer segments by the pigment epithelium.The inclusions of the pigment epithelial cells are classified in five categories which seem to be related to each other owing to their shared structural characteristics. It is suggested that melanin granules are produced, modified and destroyed by the pigment epithelial cells of the adult.Because of the relations between the photoreceptors and the melanin granules it is suggested that light scattered by the melanin granules may pass backwards through the outer segments of the cones, but not of the rods.This investigation was supported in part by the Danish Foundation for the Advancement of Science and by the Danish Medical Research Council.     

Does Melanin Turnover Occur in the Eyes of Adult Vertebrates?

This paper is a review of what is known about the turnover of melanin in iris, choroid, and retinal pigment epithelium (RPE) of the adult vertebrate eye. Differences in size and structure of choroideal and retinal pigment epithelial melanin granules are shown by electron micrographs. The classical stages of melanin synthesis, including the premelanosome, are shown in the RPE of adult hamsters that had been exposed to intense light. Degradation or synthesis of melanin also seem to occur in the melanocytes of the choroid in these animals. It is postulated that all three pigmented eye tissues (iris, RPE, and choroid) of adult vertebrates form melanin granules in vivo. However, nothing is known about the amount of this turnover.     

A newly discovered pathway of melanin formation in cultured retinal pigment epithelium of cattle

According to a recent hypothesis the melanin granules in the retinal pigment epithelium of mammals originate from photosensory membrane degradation. To test this hypothesis the retinal pigment epithelium of cattle was kept in tissue culture and exposed to gold-labelled rod outer segments. Gold granules were later detected inside phagosomes, melanosomes and mature melanin granules. Tyrosinase, the key enzyme in melanogenesis, was additionally localized inside phagosomes. These results indicate that in cultured retinal pigment epithelium the matrix of the melanosome can originate from phagosomes. therefore, the melanosome is a specialized lysosome.     

The postnatal reaction of the albino rat pigment epithelium after administration of chloroquindiphosphate]

The development of pigment granules has been studied in 0...5 day old normal albino-rats and after a single injection of chloroquinediphosphat. During the postnatal time the pigment epithelial cell changes structurally. The basal region of the epithelium containes lipoidvacuoles and later on glycogen. The apical region form long skin projections that enclose the developing inner segments of photoreceptor cells. The pigment granules develop normally until the stage of melanin deposition. The lamellated premelanosomes are equipped by helical structures. The development of premelanosomes are not disturbed, by chloroquinediphosphate blocking the protein synthesis in the cell. The drug produce lamellated vacuoles inside the cell arising from the ergastoplasm. Also the extracellular space is enlarged. A significant increase or decrease in the number of phagosomes in the pigment epithelial cells after administration of the drug does not occur as compared to the cells of untreated animals.     

Retinal development in the lamprey (Petromyzon marinus L.): premetamorphic ammocoete eye.

Development of the retina of the ammocoete begins early in embryogenesis, with the formation of the optic vesicle, but development of the rudimentary eye is suspended and remains arrested during larval life. Prior to the onset of metamorphosis, the retina of the ammocoete is completely undifferentiated, with the exception of a small area (Zone II) surrounding the optic nerve head, where all of the adult retinal layers are found. The photoreceptors in this area have developed to include synaptic contacts as well as inner and outer segments. The pigment epithelium in this area, too, has differentiated to include well-formed melanin granules, myeloid bodies and endoplasmic reticulum and is closely associated with the receptor cell outer segments. With the approach of metamorphosis, differentiation of the remainder of the retina (Zone I) begins, taking place in a radial fashion from the optic nerve head. Differentiating pigment epithelial cells adjacent to the differentiated retinal zone begin to accumulate melanin granules. In the neural retina, junctional complexes are established in the form of an external limiting membrane, and connecting cilia project into the optic ventricle. Photoreceptor differentiation begins with the formation of a mitochondria-filled ellipsoid within the inner segment. Development and differentiation of the ammocoete retina is unique to vertebrates in that only a small area of differentiated retina is present during the larval stage. The remainder of the retina differentiates and becomes functional during metamorphosis.     

Light-Induced Dopamine Release from Teleost Retinas Acts as a Light-Adaptive Signal to the Retinal Pigment Epithelium

In the retinal pigment epithelium (RPE) of lower vertebrates, melanin pigment granules migrate in and out of the cells' long apical projections in response to changes in light condition. When the RPE is in its normal association with the retina, light onset induces pigment granules to disperse into the apical projections; dark onset induces pigment granules to aggregate into the cell bodies. However, when the RPE is separated from the retina, pigment granule movement in the isolated RPE is insensitive to light onset. It thus seems likely that a signal from the retina communicates light onset to the RPE to initiate pigment dispersion. We have examined the nature of this retina-to-RPE signal in green sunfish, Lepomis cyanellus. In isolated retinas with adherent RPE, light-induced pigment dispersion in the RPE is blocked by treatments known to block Ca2+-dependent transmitter release in the retina. In addition, the medium obtained from incubating previously dark-adapted retinas in the light induces light-adaptive pigment dispersion when added to isolated RPE. In contrast, the medium obtained from incubating dark-adapted retinas in constant darkness does not affect pigment distribution when added to isolated RPE. These results are consistent with the idea that RPE pigment dispersion is triggered by a substance that diffuses from the retina at light onset. The capacity of the conditioned medium from light-incubated retinas to induce pigment dispersion in isolated RPE is inhibited by a D2 dopamine antagonist, but not by D1 or alpha-adrenergic antagonists. Light-induced pigment dispersion in whole RPE-retinas is also blocked by a D2 dopamine antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)     

光和暗对埃及蟾蜍(BUFO REGULARIS)发育中的视网膜的影响(英文)

本文研究了埃及蟾蜍(Bufo regularis Reuss)从早期幼虫到变态结束,光和暗对视网膜的影响。所观察到的对光和暗反应的变化限于色素上皮和光感受细胞。实验动物分四组:1)在亮处固定的对照(CL);2)在暗处固定的对照(CD);3)连续养在暗处的动物(DD);4)连续受光照的动物(LL)。在CD和DD组动物,黑色素颗粒在色素上皮细胞突起(PEP)中的分布限于光感受细胞顶部间之外周区(巩膜方位)。与此相反。在CL和LL组动物,大量黑色素颗粒则分散在视觉细胞外节段和椭球段间色素上皮细胞突起之向心端位置(玻璃体方位)。在这种动物,上皮细胞色素对光和暗反应发生的光机械运动出现在肢芽期以前。新变态小蟾蜍DD组眼球,与其他三组比较起来,色素上皮层相当厚并含有大的脂肪滴。在较晚期,只有DD组动物的一些杆细胞外节段呈现退化现象。其次,这一组中,由于连续缺少光照的结果,眼球之锥细胞数目有明显减少。四组动物的视网膜上也观察到锥细胞的细胞核位置略有不同。     

Ultrastructural study of the retinal pigment epithelium during metamorphosis in the sea lamprey (Petromyzon marinus L.)

Summary The sequence of morphological changes in the retinal pigment epithelium during the metamorphic period of the sea lamprey Petromyzon marinus L. has been investigated using electron microscopy. At early metamorphic stages (stages I and II), photoreceptors are present in a small zone of the retina. During these stages, the lateral surface of the epithelial cells shows zonulae occludentes and adhaerentes. The degree of cell differentiation varies throughout the retinal pigment epithelium. Cells covering the differentiated photoreceptors in the central retina have phagosomes, whereas pigment granules appear only in the retinal pigment epithelium dorsal to the optic nerve head. Most epithelial cells have myeloid bodies; their morphology is more complex around the optic nerve head. At stage III, when photoreceptors develop over the whole retina, the distribution of cytoplasmic organelles is almost homogeneous in the retinal pigment epithelium. Subsequently, the basal plasma membrane of the epithelial cells becomes progressively folded and their apical processes enlarged. In addition, extensive gap junctions develop between retinal pigment cells. In late metamorphic stages, noticeable growth of myeloid bodies occurs and consequently the retinal pigment epithelium resembles that of the adult. This study also describes, for the first time, the presence of wandering phagocytes in the retinal pigment epithelium of lampreys; their role in melanosome degradation is discussed.     

Stimulation of Distinct D2 Dopaminergic and α2-Adrenergic Receptors Induces Light-Adaptive Pigment Dispersion in Teleost Retinal Pigment Epithelium

In the retinal pigment epithelium (RPE) of lower vertebrates, melanin pigment granules aggregate and disperse in response to changes in light conditions. Pigment granules aggregate into the RPE cell body in the dark and disperse into the long apical projections in the light. Pigment granule movement retains its light sensitivity in vitro only if RPE is explanted together with neural retina. In the absence of retina, RPE pigment granules no longer move in response to light onset or offset. Using a preparation of mechanically isolated fragments of RPE from green sunfish, Lepomis cyanellus, we investigated the effects of catecholamines on pigment migration. We report here that 3,4-dihydoxyphenylethylamine (dopamine) and clonidine each mimic the effect of light in vivo by inducing pigment granule dispersion. Dopamine had a half-maximal effect at approximately 2 nM; clonidine, at 1 microM. Dopamine-induced dispersion was inhibited by the D2 dopaminergic antagonist sulpiride but not by D1 or alpha-adrenergic antagonists. Furthermore, a D2 dopaminergic agonist (LY 171555) but not a D1 dopaminergic agonist (SKF 38393) mimicked the effect of dopamine. Clonidine-induced dispersion was inhibited by the alpha 2-adrenergic antagonist yohimbine but not by sulpiride. These results suggest that teleost RPE cells possess distinct D2 dopaminergic and alpha 2-adrenergic receptors, and that stimulation of either receptor type is sufficient to induce pigment granule dispersion. In addition, forskolin, an activator of adenylate cyclase, induced pigment granule movement in the opposite direction, i.e., dark-adaptive pigment aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)     

The human vocal cord surface

Summary The effect of phenylthiourea (PTU) on the pigment epithelium and the photoreceptor cells in the developing retina of Haplochromis burtoni was studied by electron microscopy. In the retinal pigment epithelium of 6-day old embryos, both types of melanin granule (spindle-shaped and rod-shaped) are already found. PTU inhibits the biosynthesis of melanin but does not influence the formation of premelanosomes so that in PTU-treated embryos there are no melanosomes, but an abundance of premelanosomes. The structure of the premelanosomes is described. It differs completely from that of all other vertebrates. Other changes: an increase in polysomes, retarded development of the inner segment of the photoreceptor cells and enlargement of the intercellular space between the inner and outer leaflet of the retina, may be due to a toxic effect of PTU.This investigation was supported by grants of the Deutsche Forschungsgemeinschaft     

The classical pathway of melanogenesis is not essential for melanin synthesis in the adult retinal pigment epithelium

Premelanosomes are presumed to be essential for melanogenesis in melanocytes and pre-natal retinal pigment epithelium (RPE) cells. We analysed melanin synthesis in adenoviral-transduced tyrosinase-gene-expressing amelanotic RPE (ARPE) 19 cells to determine whether premelanosome formation is needed for post-natal melanogenesis. The synthesis of melanogenic proteins and melanin granules was investigated by immunocytochemistry and light and electron microscopy. The occurrence of tyrosinase was analysed ultrastructurally by dihydroxyphenylalanine histochemistry. The viability of transduced cell cultures was examined via MTT assay. We found active tyrosinase in small granule-like vesicles throughout the cytoplasm and in the endoplasmic reticulum and nuclear membrane. Tyrosinase was also associated with multi-vesicular and multi-lamellar organelles. Typical premelanosomes, structural protein PMEL17, tyrosinase-related protein 1 and classic melanosomal stages I–IV were not detected. Instead, melanogenesis took place inside multi-vesicular and multi-lamellar bodies of unknown origin. Viability was not affected up to 10 days after transduction. We thus demonstrate a pathway of melanin formation lacking typical hallmarks of melanogenesis.     

Molecular Characterization of the Mouse Tyrosinase Gene:Pigment Cell-Specific Expression in Transgenic Mice

Tyrosinase is the key enzyme in melanin synthesis, and is expressed in the pigment epithelium of the retina, a cell layer derived from the optic cup; and in neural crest-derived melanocytes of skin, hair follicle, choroid, and iris. The tyrosinase gene has been cloned and shown to map to the well-characterized c-locus (albino locus) of the mouse. Subsequent studies demonstrated that a functional tyrosinase minigene was able to rescue the albino phenotype in transgenic mice. The transgene was expressed in a cell type-specific manner in skin and eye. During development of the mouse, the tyrosinase gene is expressed in the pigment epithelium of the retina as early as day 10.5 of gestation. In the hair follicle, tyrosinase gene expression is detected from day 16.5 onwards. This cell-type–specific expression is largely reproduced in transgenic mice. Our results suggest that sequences in the immediate vicinity of the mouse tyrosinase gene are sufficient to provide cell type-specificity and developmental regulation in melanocytes and the pigment epithelium.     

Development of Pigment Cells in the Brain of Ascidian Tadpole Larvae: Insights into the Origins of Vertebrate Pigment Cells

In vertebrates, melanins produced in specialized pigment cells are required for visual acuity, camouflage, sexual display and protection from ultra violet (UV) radiation. There are three pigment cell types that are classified based on their distinct embryonic origins. Retinal pigment epithelium (RPE) cells originate from the outer layer of the optic cup. Pigment cells of the pineal organ are formed from the developing diencephalon. Melanocytes are derived from the neural crest unique to vertebrate embryos. Some of these pigment cells also play roles that are independent of the activity of tyrosinase, the key melanogenesis enzyme, or melanin: production of substrate(s) for catecholamine synthesis, maintenance of endolymph composition in the cochlea, maintenance of photoreceptor cells in the retina and retinoid metabolism essential for the visual cycle. To deduce the evolutionary origins of vertebrate pigment cells and a possible archetypal genetic circuitry, which may have been modified and utilized to generate multiple pigment cell types, comparison of developmental mechanisms of pigment cells between vertebrates and closely related invertebrate ascidians are proposed to provide useful information. The tadpole‐type larva of ascidians possesses two melanin‐containing pigment cells, termed the otolith and ocellus pigment cells, in the brain that are believed to be required for photo‐ and geotactic responses during swimming. In this review, current knowledge on the development of the two ascidian pigment cells is summarized, i.e. complete cell lineage, structure and expression of genes encoding two melanogenesis enzymes, and molecular developmental mechanisms involving BMP‐CHORDIN antagonism, and possible evolutionary relationships between ascidian and vertebrate pigment cells are discussed.     

Ultimate Fate of Rod Outer Segments in the Retinal Pigment Epithelium

To investigate the degradation pathway of rod outer segments (ROS) in vivo, we injected gold-labeled ROS into the subretinal space of rabbits using a pars plana approach. Histology and electron microscopy performed on the specimens 72 hr after ROS injection revealed that the retina over the injection site was reattached, the retinal pigment epithelial (RPE) cells were intact, and gold granules were localized inside melanin granules and melanosomes. These results indicate that, in RPE, in vivo degradation of ROS is associated with melanosomes.     

Ultimate fate of rod outer segments in the retinal pigment epithelium.

To investigate the degradation pathway of rod outer segments (ROS) in vivo, we injected gold-labeled ROS into the subretinal space of rabbits using a pars plana approach. Histology and electron microscopy performed on the specimens 72 hr after ROS injection revealed that the retina over the injection site was reattached, the retinal pigment epithelial (RPE) cells were intact, and gold granules were localized inside melanin granules and melanosomes. These results indicate that, in RPE, in vivo degradation of ROS is associated with melanosomes.     

Morphogenesis and nature of the pigment granules in the adult human retinal pigment epithelium

Summary The pigment epithelial cells of the retina are a layer of highly specialized melanocytes. Beginning in the early embryonic period they produce melanin throughout the entire life. The Golgi apparatus plays a key role in the biosynthesis of melanin. The following steps can be distinguished morphologically: (a) Golgi-vesicles, (b) intermediate vesicles, (c) melanosomes, (d) melanin granules. Structures with a ringlike appearance that are described as lipofuscin granules in the literature prove to be altered intermediate vesicles and melanosomes.This investigation was carried out in part at the Francis I. Proctor Foundation for Research in Ophthalmology, San Francisco, California, U.S.A., and supported by United States Public Health Service Program Project Grant EY 00310, and Deutsche Forschungsgemeinschaft, Training Grant Nr. Sp 102/1.     

Effects of melatonin and light on porphyrin synthesis in the bovine retina, pigment epithelium and choroid.

The quantity of porphyrin synthesized in the presence of 10(-3) M delta-aminolevulinic acid (ALA) is several times higher in the bovine pigment epithelium than in the retina. Synthesis in the retina was found to be increased by illumination, whereas synthesis in the pigment epithelium was decreased if the whole anatomical unit (retina-pigment epithelium-choroid) was cultivated together. The quantity of porphyrin synthesized in the presence of 10(-3) M ALA or 10(-6) M melatonin was different when the pigment epithelium and retina were separated. The combination 10(-3) M ALA with 10(-6) M melatonin inhibited retinal porphyrin synthesis after green light adaptation, while in the pigment epithelium green light adaptation induced porphyrin synthesis. It is postulated that the light-sensitive porphyrin-haeme synthesis of the retina-pigment epithelium-choroid functional unit may serve and modulate the synthesis of guanylate cyclase for cGMP.     

Detection of a Fine Lamellar Gridwork After Degradation of Ocular Melanin Granules by Cultured Peritoneal Macrophages

In the present study we investigated by electron microscopy whether melanin granules derived from choroidal melanocytes and retinal pigment epithelium of cattle could be degraded in the phagolysosomes of cultured murine macrophages. It was found that degradation of ocular melanin is possible by the lysosomes of these macrophages. During degradation of the melanin granules an internal gridwork of fine concentric, highly ordered membranes, 3-4 nm thick, became visible. These membranes may represent remnants of the melanin polymer in the original melanosome or may result from self-assembly of degradation products. Early-stage melanosome-like structures also appeared during digestion of these melanin granules. Melanin granules that seemed to break down into smaller fragments without any visible internal structure were also observed.