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PubMed2003

A novel Xenopus SWS2, P434 visual pigment: structure, cellular location, and spectral analyses.

PURPOSE: The purpose of this study was to clone and characterize the green rod pigment in Xenopus laevis. METHODS: The cDNA for the Xenopus "green rod" pigment was cloned and sequenced from Xenopus retina mRNA by reverse transcription polymerase chain reaction and the 5' end cloned by rapid amplification of the cDNA ends. The cellular localization of the Xenopus opsin was determined by immunolabeling of flat-mounted retinas using a specific antibody against this opsin. Spectral properties of the expressed protein were determined by absorption spectroscopy using recombinant pigment. RESULTS: A novel Xenopus opsin cDNA containing a full-length coding region has been cloned and sequenced. The deduced amino acid sequence predicts a protein of 362 amino acids, forming 7 hydrophobic helices. Sequence analysis indicates that it belongs firmly to the SWS2 class of visual pigments and has 89%, 80%, and 75% amino acid sequence identity with bullfrog, tiger salamander, and newt SWS2 pigments, respectively. Staining of Xenopus retina with a Xenopus SWS2 opsin-specific polyclonal antibody demonstrated that the SWS2 pigment is expressed in green rods. After expression in COS cells, reconstitution with 11-cis retinal, and purification, the SWS2 pigment exhibits an absolute absorption maximum of 434 nm Thus, the name "SWS2, P434" was assigned for this opsin. The pigment decays rapidly in hydroxylamine in the dark, unlike the red rod pigment, rhodopsin. CONCLUSIONS: A novel green rod opsin cDNA has been cloned and sequenced from the retina of adult Xenopus laevis, which encodes a protein belonging to the SWS2 group of opsins. The expressed opsin possesses cone-opsin-like properties although it was identified only in the Xenopus green rod cells.

Amino Acid Sequence↗
PubMed1997

Intraleucocyte malaria pigment in asymptomatic and uncomplicated malaria.

While malaria pigment or haemozoin is known to be an end product of haemoglobin digestion by the malaria parasite, its clinical significance is just beginning to be elucidated. We have studied the distribution of intraleucocyte malaria pigment in 92 children, consisting of 32 children with asymptomatic malaria, 32 children with mild or uncomplicated malaria and 28 children with no malaria. Over 90% of children in each of the three groups had pigment-containing monocytes and the numbers of pigment-containing monocytes were not significantly different between the three groups. While over 90% of children in both the asymptomatic malaria and uncomplicated malaria groups had pigment-containing neutrophils, 71.4% of the no malaria group had such neutrophils. The numbers of pigment containing neutrophils was highest in the uncomplicated malaria group, followed by the asymptomatic malaria group with the no malaria group having the least numbers. The pigmented neutrophil: monocyte ratio followed the same pattern. It was concluded that the number of pigment-containing neutrophils and the pigmented neutrophil:monocyte ratio may be a marker of the severity of malaria infection when one considers the conditions: no malaria, asymptomatic malaria and mild malaria. Further work to verify this hypothesis across the full spectrum of the manifestations of malaria infection is needed.

Adolescent↗
PubMed2004

Autofluorescence method to measure macular pigment optical densities fluorometry and autofluorescence imaging.

Non-invasive measurement of the optical density of the human macular pigment by the autofluorescence method takes advantage of the fluorescence of lipofuscin in the human retinal pigment epithelium. Measuring the intensity of fluorescence above 550 nm, where macular pigment has essentially zero absorption, and stimulating the fluorescence with two wavelengths, one well absorbed by macular pigment and the other minimally absorbed by macular pigment, provides a single-pass measurement of the macular pigment optical density. The method is implemented either by fluorometry of lipofuscin to yield the optical density of the macular pigment in a 2 degrees diameter central area, or by autofluorescence imaging to yield a high-resolution map of the macular pigment distribution.

Female↗
PubMedDate not supplied

Neuromodulation of pigment movement in the RPE of normal and 6-OHDA-lesioned goldfish retinas.

The role of dopamine as the endogenous signal-initiating light-dependent changes in the distribution of pigment granules in goldfish retinal pigment epithelium was investigated. In normal retinas, light adaptation resulted in the dispersion of pigment granules. This effect of light was mimicked by the intraocular injection of dopamine or serotonin, which is thought to increase endogenous dopamine release, into dark-adapted eyes. The effect of light, dopamine, or serotonin on dark-adapted retinas was blocked by the dopamine receptor antagonists haloperidol and sulpiride. However, lesioning the endogenous source of retinal dopamine, by prior intraocular injection of 6-hydroxydopamine (6-OHDA), did not block the dispersion of pigment granules in light-adapted retinas. No significant differences in pigment dispersion were noted between unlesioned and lesioned light- or dark-adapted retinas. However, the effect of light on pigment dispersion was no longer blocked by haloperidol or sulpiride in 6-OHDA lesioned animals. Dopamine and serotonin mimicked the effect of light when injected into lesioned dark-adapted eyes, but their effects were also not blocked by haloperidol or sulpiride. These results suggest that dopamine, acting on D2 receptors, is sufficient to induce pigment migration in unlesioned animals. In 6-OHDA-lesioned animals, however, pigment migration is mediated by a receptor mechanism other than D2.

Animals↗
PubMed1963

THE PHOTOSENSITIVE RETINAL PIGMENT SYSTEM OF GEKKO GEKKO.

Retinal extracts of Gekko gekko were found to contain two retinene(1) photopigments, one with maximum absorption at about 521 mmicro, the second with a maximum in the region of 478 mmicro. These pigments were assayed by the method of partial bleaching and their spectral characteristics studied by examining their difference spectra. The 478 mmicro pigment was present in the extracts as 8 per cent of the total photopigment concentration. The two pigment systems were shown to be biochemically independent and to have different properties. Unlike the 521 mmicro pigment, for example, the 478 mmicro pigment was found to resist the action of NH(2)OH and, within the cells, to be unaffected by sucrose solutions. These solutions destroyed or altered the 521 system so that extracts of sucrose-treated retinae were found to contain significantly less 521 photopigment. In digitonin solution the 521 pigment was unaffected by sucrose treatment. Both pigments were extracted from separated, washed outer segments and so are considered to be visual pigments. This dual system accounts for the spectral sensitivity of this gecko as determined by Denton. A search was made, but no evidence was secured for the presence of a photopigment absorbing at longer wavelengths. Electoretinographic data suggest, however, that an elevated sensitivity at longer wavelengths occurs in some geckos so that a continued search is justified for a third photopigment.

Animals↗
PubMedDate not supplied

Eye color pigment granules in Drosophila mauritiana: mosaics produced by excision of a transposable element.

Compound eyes of the white-peach (wpch) mutant strain of Drosophila mauritiana have some pigment and receptor cells with wild-type eye color pigmentation. These eyes are mosaic, because excision of a transposable element reverts wpch to wild type during the development of somatic cells. Wild-type patches have three types of pigment granule residing in three respective cell types: primary pigment cells, secondary pigment cells, and retinula (visual receptor) cells. Most aspects of these granules, as well as all other aspects of compound eye ultrastructure, are exactly as in the better studied sibling species D. melanogaster. In the wpch parts of the eye, small and giant unpigmented "pigment granules" reside in secondary pigment cells. These white granules are just like the corresponding granules of w mutant D. melanogaster. Small vs. large patches of pigmented cells likely represent excision events occurring late vs. early respectively during development. Mosaics of eye color markers have been important in developmental analyses; the ease of constructing mosaics of D. mauritiana gives this preparation advantages for mosaic analyses.

Animals↗
PubMed1971

Pigmentation and acriflavine resistance in Serratia marcescens.

Stable, orange, acriflavine-resistant variants were selected by treatment of a wild-type, red, acriflavine-sensitive strain of Serratia marcescens with acriflavine. Visible, ultraviolet, infrared, and nuclear magnetic resonance spectra of purified pigment from the red strain were identical to those of the pigment from the orange strain, and the orange mutant was not due to a mutation affecting the structure of the pigment, prodigiosin. The color of the red strain was not affected by variations in pH between 5.0 and 8.0, whereas the color of the orange mutant changed from pink to orange over the same pH range. This variation was mimicked by the pH-induced variation in color of prodigiosin purified from either the red, wild-type or the orange, mutant strains. Density-gradient centrifugation of cell fragments after ultrasonic disintegration resulted in characteristic pigmented bands. Biochemical characterization of these pigmented bands showed that they contained pigment and a protein component, but no lipids, polysaccharides, sugars, glucosamine, or phosphates were detected. Further fractionation of these pigmented bands by zone electrophoresis on a sucrose density gradient indicated that some pigment in S. marcescens was specifically attached to protein components.

Acridines↗
PubMed1986

The effects of dim cyclic light on pigment epithelial function in the albino rat.

Most pathologies of the outer retina include physiological and morphological changes in the pigment epithelium. The question of pigment epithelial involvement in retinal light damage caused by low intensities of light is still unresolved. In the present study, we investigated the effects of low intensity cyclic light on pigment epithelial function in albino rats. The functioning of the pigment epithelium was assessed electrophysiologically from d.c. recordings of ERG c-waves and sodium azide induced changes in the resting potential. Responses obtained from albino rats raised under low intensity cyclic light (0.63 ft cd. 12:12 L:D) were compared to those obtained from albino rats raised under minimal light exposure conditions (dark-reared) and pigmented rats housed under low intensity cyclic light. We report, for the first time, that albino rats raised from birth under low intensity cyclic light possess c-waves. Their responses were comparable in amplitude and latency to those recorded from pigmented rats housed under similar conditions, but were significantly smaller than those recorded from dark-reared albino rats. The reduction in the amplitudes of the c-waves recorded from cyclic light-reared albino rats was probably not due to retinal light damage. Comparisons of the amplitudes and latencies of ERG b-waves recorded from cyclic light-reared and dark-reared albino rats did not suggest that the retinas of the cyclic light-reared albino rats had been damaged by light. Light microscopic examination of these retinas also provided no evidence for light damage. The transient, positive potential changes recorded from cyclic light-reared albino rats in response to bolus injections of sodium azide were significantly smaller than those recorded from either dark-reared albino rats or pigmented rats housed under low intensity cyclic light. The results of these experiments suggest that the pigment epithelium of albino rats is functionally altered by extremely low intensities of cyclic light.

Albinism↗
PubMed1985

[Pigmented lesions of the oral cavity].

Pigmented lesions of the oral cavity are of multiple origin. They can be subdivided as follows: non tumoral pigmentations, non melanin pigmented tumors or tumor-like lesions, benign melanin pigmented tumors and malignant melanomas. Among non tumoral pigmented lesions, some of them show melanin deposits: they can be associated with a systemic disease (Peutz Jeghers syndrome, Addison's disease) or have a medicamentous origin, or belong to a lichen migricans. Non tumoral and non melanin pigmentations are principally due to a heavy metal accumulation or an accidental tatoo arising after tooth treatment. Peripheral giant cell granuloma, so-called giant cell epulis is the major non pigmented non melanin pseudotumoral lesion; pigmentation is due to hemosiderin deposits. In the oral cavity nevi are principally of the intramucosal type. Blue nevus, the second type in frequency, is usually located on the hard palate. Primary malignant melanomas are rare in the oral cavity, but it is--because its very bad prognosis--the most important lesion. In order to improve the survival it is necessary to do the diagnosis as early as possible.

Adult↗
PubMed1978

Changes of the external and internal pigment pattern upon fertilization in the egg of Xenopus laevis.

External and internal pigment shifts in Xenopus laevis eggs were studied between fertilization and first cleavage. Externally visible, constant features are: (1) the 'activation contraction', a pigment shift towards the animal side taking place between 5 and 15 min post fertilization (p.f.) and (2) the concentration of the pigment around the sperm entrance point leading to the formation of the grey crescent at the opposite side of the egg. Hence, in Xenopus the grey crescent is not formed by rotation of the pigmented cortical layer with respect to the internal egg mass. Histological examination reveals that during the activation contraction the pigment is mainly concentrated in the cortical cytoplasm. Except in the region around the sperm entrance point, from 15 min p.f. onwards, the pigment progressively disperses through the subcortical layer and part of it even moves more deeply into the egg. After fusion of the pronuclei (45--60 min p.f.) the pigment in the subcortical layer forms aggregates. During the pigment shift the yolk-free cytoplasm is displaced dorsally and is ultimately found opposite the sperm entrance point. Thin fibrillar structures in the yolk-free cytoplasm progressively orient themselves parallel to the dorso-ventral plane, and from 40 min p.f. onwards towards the pronuclei. These observations are discussed in connexion with cinematographic observations by Hara, Tydeman & Hengst (1977).

Animals↗
PubMed1995

In vivo observation of magnified features of pigmented lesions on volar skin using video macroscope. Usefulness of epiluminescence techniques in clinical diagnosis.

BACKGROUND AND DESIGN: In vivo epiluminescence microscopy is now used as a useful noninvasive method for determining clinical diagnosis of pigmented skin lesions. Until now, however, pigmented lesions on the volar skin have been hardly studied with this method. In the present epiluminescent study, various kinds of pigmented lesions on the volar skin were extensively investigated by means of video macroscope, a newly developed electronic device with a higher magnification power, and correlation between the magnified features and histopathologic findings was evaluated. RESULTS: Magnified features of most lesions of acquired or congenital melanocytic nevus on the volar skin were classified into the following three typical patterns: (1) a parallel pattern formed by pigmented parallel lines corresponding to the furrows of the skin markings, (2) a latticelike pattern composed of pigmented lines along and across the furrows of the skin markings, and (3) a fibrillar pattern formed by densely packed, fibrillar pigmented lines arranged in the direction crossing the furrows. In contrast, macular or plaque portions of acral lentiginous melanoma exhibited disorderly arranged, irregular pigment patterns, mainly affecting the ridges of the skin markings. In addition, brown globules of various shades and many black dots of variable sizes were often observed and, on the margin of the lesions, pseudopods and/or the "serrated" pattern were detected. Cutaneous hemorrhagic macule and so-called black heel showed highly specific features and thus could be easily diagnosed with video macroscopy. CONCLUSION: Video macroscope proved to be a very useful instrument for the diagnosis of pigmented lesions on the volar skin.

Equipment Design↗
PubMed1989

Does pigmentation affect the trabecular meshwork?

Pigment has been associated with glaucoma in pigment dispersion syndrome and in intraocular lens-associated pigment dispersion. Human eyes with segmental pigmentation of the trabecular meshwork may provide a controlled model to study the effects of pigment on the meshwork. Nine normal human eye bank eyes with segmental pigmentation of the trabecular meshwork were examined for pigment-associated differences in trabecular cellularity or morphology. Cellularity was examined with four methods: cells per histologic section, cells per length of tissue, cells per overall area, and cells per solid tissue. The simplest method, cells per histologic section, gave results equivalent to the other methods. No differences in cellularity or morphology were found between pigmented and nonpigmented areas.

Adult↗
PubMed1999

Evolution of visual pigments and related molecules.

The molecular phylogenetic tree of vertebrate visual pigments, constructed on the basis of amino acid sequence identity, suggests that the visual pigments can be classified into five groups (L, ML, MS, S and Rh) and that their genes have evolved along these five gene lines. Goldfish has a UV-sensitive visual pigment (S group) localized in miniature single cone cells. Medaka has one type of rod cell containing rhodopsin (Rh group) and four types of cone cells, each of which contains a specific visual pigment with an absorption maximum that differs from those of the others. Frogs have a violet-sensitive visual pigment (S group) in small single cone cells and a blue-sensitive visual pigment (MS group) in green rod cells. Although nocturnal and diurnal geckos have rod- and cone-based retinas, respectively, they have phylogenetically closely related visual pigments. The pigments in each line may have restricted absorption maxima. We have cloned cDNAs encoding molecules involved in the phototransduction system of visual cells, such as phosphodiesterase, opsin kinase and arrestin. We then constructed phylogenetic trees of these molecules with the deduced amino acid sequences. The resulting phylogenetic trees show that these molecules are classified into two groups; one is expressed in cones and another in rods, suggesting that rods and cones contain homologous molecules with different amino acid sequences. These differences may result in the different light responses of rods and cones.

Animals↗
PubMed1995

Expression of kinesin heavy chain isoforms in retinal pigment epithelial cells.

To examine the possible role of kinesin in pigment granule migration in the retinal pigment epithelium (RPE) of teleosts, we investigated the expression and distribution of kinesin heavy chain (KHC) in RPE. Blots of fish RPE lysates probed with two well-characterized antibodies to KHC (H2 and HD) displayed a prominent band at 120 kD. A third KHC antibody (SUK4) recognized a band at 118 kD. The 118 kD band was also occasionally present in blots probed with H2, suggesting the presence of two KHC isoforms in teleost RPE. Reverse transcriptase-polymerase chain reaction (RT-PCR) of mRNA from RPE using primers homologous to conserved regions of the KHC motor domain resulted in the identification of two putative KHC genes (FKIF1 and FKIF5) based on partial amino acid sequences. Previous studies had demonstrated a requirement for microtubules in pigment granule aggregation in RPE. In addition, the reported microtubule polarity orientation in RPE apical projections is consistent with a role for kinesin in pigment granule aggregation. Immunofluorescent localization of KHC in isolated RPE cells using H2 revealed a mottled distribution over the entire cell body, with no detectable selective association with pigment granules, even in cells fixed while aggregating pigment granules. Microinjected KHC antibodies had no effect on pigment granule aggregation or dispersion, although each of the three antibodies has been shown to block kinesin function in other systems. Thus we found no evidence for KHC function in RPE pigment granule aggregation. However, the two KHC isoforms may participate in other microtubule-dependent processes in RPE.

Amino Acid Sequence↗
PubMed1981

Axonal guidance during development of the optic nerve: the role of pigmented epithelia and other extrinsic factors.

It is well established that a congenital lack of ocular melanin (albinism) can lead to developmental abnormalities of the central visual pathways. However, it is yet unknown how the pigmentation per se acts to influence formation of the optic projection. In order to study the possible interaction between eye pigment and optic axons during development, we have examined, with the use of serial section techniques, a series of timed embryos at stages when the ocular pigment and outgrowing axons first become apparent. Our results have demonstrated that, in mice and rats, the upper wall of the distal half of the primitive eye stalk (a region which lies along the potential route to be taken by the earliest developing nerve fibers) is transiently pigmented prior to and during the migration of the pioneer optic axons. All outgrowing neurites avoid this stretch of melanotic tissue and instead grow preferentially through a system of extracellular tunnels in the ventral, pigment-free zones of the distal eye stalk. The stalk remains unpigmented from about its midpoint and continuing toward the brain. At the pigment/pigment-free interface many of the axons shift upward from their ventral positions, forming a marginal annulus. In the chick, on the contrary, pigmentation of the stalk does not occur and as the optic axons exit the globe they grow immediately in an annulus configuration. In Xenopus, the entire stalk becomes pigmented and the optic fibers congregate in one discrete bundle of fascicles along the length of the stalk's most ventral margin. These observations suggest that melanin-producing stalk cells may play a role in controlling the topographic patterning of optic fibers within the developing nerve by inhibiting the lateral spread of axonal growth cones into or within their territory. To test this hypothesis we have charted the distribution of optic fibers in the developing optic stalks of timed albino rat embryos. Indeed, as fibers leave the mutant eye, it was found that a small but consistent number of pioneering axons (day E15) become ectopic and immediately invade nonpigmented regions (those normally pigmented and axon-free) in the distal optic stalk. Thus, the usual topographic arrangement of the collection of pioneer optic fibers is altered in the albino.

Animals↗
PubMed1988

Aberrant optic axons in the retinal pigment epithelium during chick and quail visual pathway development.

Examination of a large number of retinal pigment epithelia revealed that, in a small proportion, optic axons in chick and quail eyes aberrantly entered the pigment cell layer between embryonic day (E) 7 to E14. The aberrant retinal axons originated from the main stream of retinal fibers in the optic nerve and invaded the pigment layer from various positions of the optic nerve head or fissure by growing along the basal side of the pigment epithelium. The axon bundles grew several millimeters into the epithelial sheet and arborized at the margin of the eye. As shown by electron microscopy the nerve fibers occurred as bundles of three to several hundred axons. They always were located at the basal side of the epithelium, and were enveloped by processes of epithelial cells. Very large bundles of axons, however, displaced the epithelial cells from the basal matrix. These retinal axons contacted the pigment epithelial basal lamina. The basal extracellular matrix from the retinal pigment epithelium was isolated and used as substratum for in vitro cultures of various types of neural explants. The matrix preparations consisted of a sheet of a 50 nm thick basal lamina with a central lamina densa, two laminae rarae, and a 15 micron thick stroma. Axons from avian retina explants, as well as sensory ganglia, grew on the basal lamina side of the pigment cell matrix with the same growth rate and with the same fiber density as on similarly prepared basal laminae from the neural retina. These experiments show that the matrix from the pigment epithelium of the avian eye does not have negative effects on axonal growth and indicate that a basal lamina from a normally non-innervated tissue can provide a favorable matrix for axonal growth.

Animals↗
PubMed1996

Methionine, but not taurine, protects against formation of canine pigment gallstones.

Dogs fed a marginal protein, high carbohydrate diet containing borderline amounts of methionine consistently develop pigment gallstones, marked taurine deficiency, and abnormal secretion of unconjugated bile salts. Since taurine is essential for normal secretion of bile salts in the dog, a species that cannot use glycine for bile salt conjugation, we hypothesized that taurine deficiency plays an important role in the pathogenesis of canine pigment gallstones. The rates of formation of pigment gallstones at 6 weeks were compared in dogs fed normal dog chow, lithogenic diet alone, and lithogenic diet supplemented with either taurine or methionine (45-55 mg/kg/day). No dog fed normal dog chow, but 11 of 12 dogs fed lithogenic diet, formed pigment gallstones and sludge. Supplementation of the lithogenic diet with taurine did not protect against pigment gallstones; five of six dogs developed gallstones and sludge. However, supplementation with methionine was protective, only one of six dogs was found to have a few flecks of pigmented material in its gallbladder. Interestingly, neither taurine nor methionine protected against previously observed increases in the concentrations of biliary calcium and bile salt profile indicating that at least some of the abnormalities previously observed in this model are not related to methionine/taurine deficiency. These results disprove our initial hypothesis that taurine deficiency plays an important role in the pathogenesis of pigment gallstones, but did show that a lack of methionine is linked to the formation and growth of canine pigment gallstones. The mechanism(s) leading to the formation of gallstones in dogs that are methionine deficient are is not clear from these studies.

Animals↗
PubMed1992

Pigment epithelial repair.

We studied the repair of large, surgically induced retinal pigment epithelial defects in rabbit eyes for up to 28 days. Ophthalmoscopically, the pigment epithelium surrounding the defect became less pigmented and scattered pale foci appeared. The margin remained distinct for 28 days and the defect was subsequently covered by a variable lacework of pigmentation. Angiographically, the acute defect was hyperfluorescent and leaked heavily, but within 7 days fluorescein leakage had ceased. Histology, transmission and scanning electron microscopy, and autoradiography for the study of cells in division revealed that the pigment epithelium surrounding the defect became pleomorphic after 1 day, began to proliferate within 3 days, and covered the defect within 7 days. Subsequently, many cells became pigmented and some were fibrocyte-like. The cells within the healing defect appeared to have derived from the pigment epithelium. Our findings suggest that similar processes occur in large human defects such as pigment epithelial rips.

Animals↗