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At least 775 records · Page 43Linked to original sources
PubMed2001

Microenvironmental regulation of visual pigment expression in the chick retina.

Visual pigment (VP) expression in the chick embryo retina was investigated in ovo, in dissociated and explant cultures, and in cDNAs from individual cells. While VP mRNA is not detectable by in situ hybridization until embryonic day (ED) 14-16 in ovo, analysis of VP expression by RT-PCR showed that VP messages are present in the retina as many as 7-10 days before they become detectable by in situ hybridization, and are also detected in other regions of the embryonic CNS. On the other hand, red opsin expression is markedly accelerated when cells are isolated from their intraocular microenvironment at ED 6, and placed in pigment epithelium-free dissociated or explant cultures. This acceleration occurs regardless of cell density, birth date, or serum presence in the medium, suggesting that many photoreceptors are already programmed to express red opsin on or before ED 6, and that microenvironmental inhibitory factors prevent implementation of this program until ED 14 in ovo. The selectivity of this phenomenon is suggested by the finding that other VPs are not observed by in situ hybridization in ED 6 cultures, although they are detectable in cultures of older retinas. Taken together, these findings suggest that red opsin expression may be constitutive for many developing photoreceptor cells in the chick.

Animals↗
PubMed1994

Light deprivation profoundly affects gene expression of interphotoreceptor retinoid-binding protein in the mouse eye.

Ambient light appears to play a role in regulating gene and protein expression of interphotoreceptor retinoid-binding protein (IRBP), a protein that facilitates the transport of retinoids between the neural retina and pigment epithelium in the visual cycle. Pregnant CD-1 mice were placed in the dark approximately 48 hr before parturition, and the pups were reared for 14 days under these conditions. Control animals were reared on a 12 hr light/12 hr dark cycle. Northern blotting of total RNA isolated from whole mouse eyes at post-natal days 7-14 (P7-P14) showed a marked reduction in IRBP message in the light-deprived animals to 10-20% of levels in control animals. Reprobing of the blots for opsin and S-antigen message showed a significant decrease of about 80-90% in opsin message at 5.1 kb but no change in S-antigen message in the eyes of light-deprived mice. Light microscopic examination of the light-deprived mouse retinas showed no apparent abnormalities in morphological development and immunocytochemistry demonstrated normal distribution and levels of IRBP protein. Immunochemical quantitation of IRBP protein confirmed that there was no reduction in light-deprived as compared to normal mouse eyes. Similarly, when adult mice were light-deprived for 14 days, a marked reduction in IRBP message was also observed with no decrease in the amount of IRBP protein. Thus, light deprivation causes a large decrease in IRBP message in the mouse eye, but IRBP protein is not decreased. The dramatic effect of light deprivation on IRBP mRNA and some opsin mRNAs, but not on S-antigen message and the fact that IRBP protein levels are relatively unaffected, suggest a complex pathway of light regulation of photoreceptor function previously not encountered. This may involve regulatory controls at levels including gene transcription, mRNA stability or protein degradation that may make use of a feedback control mechanism involving light- or dark-dependent signal transduction.

Animals↗
PubMed1996

Selective absence of cone outer segment beta 3-transducin immunoreactivity in hereditary cone degeneration (cd).

We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In the cd retina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ hybridization with a bovine rod opsin and human red/green cone opsin cRNA probes, or were reacted with antibodies directed against photoreceptor-specific proteins and visualized with appropriate biotinylated antibodies. Antibodies against the following proteins were used: alpha- and beta 3-transducins, phosducin, alpha/beta- and gamma-phosphodiesterases, COS-1, and OS-2, opsin, S-antigen and IRBP. Immunoreactivity or hybridization labeling was evaluated in unstained sections; cone pathology was judged in adjacent Toluidine Blue-stained sections. With these methods it was possible to evaluate immunoreactivity or hybridization labeling and cone pathology at the single cell level. Both middle-(COS-1) and short-(OS-2) wavelength-sensitive cones were present in controls and cd affected retinae at 2.2 months, and distinct transcripts of the red/green cone pigment gene were identified in the majority of cones in both normal and affected retinas at this age. However, beta 3-transducin immunoreactivity was completely absent from cd-affected cone outer segments. Both cone types were present but in reduced numbers in older animals (11.5 and 17 months), and no reactivity to beta 3-transducin was noted. No differences were found with the other antibodies used. The specific absence of beta 3-transducin immunoreactivity from the cone outer segments suggests a potential involvement of the beta 3-transducin gene or gene product in the disease process.

Animals↗
PubMed1999

An immunohistochemical study of an autosomal dominant feline rod/cone dysplasia (Rdy cats).

An autosomal dominant, early onset feline model of rod/cone dysplasia has been described. The clinical features, light and electron microscopy, and the electrophysiology were documented. We have now examined in more detail the histopathological and immunohistochemical changes during the early phase of the disease using antibodies against opsin, synaptophysin, glial fibrillary acidic protein (GFAP) and an epithelial marker (MNF118). We have also demonstrated programmed cell death by a modified TUNEL (Terminal deoxynucleotidyl transferase, Uridine triphosphate, Nick End Labelling) technique. In the Rdy cats, there was significant photoreceptor degeneration between 5 and 17 weeks of age. The TUNEL-labeled cell and pyknotic cell counts in the outer nuclear layer peaked at around 9 weeks of age. Accumulation of opsin in the entire outer nuclear layer of the retina was noted with opsin-immunolabeled rod neurite sprouting. There was a reduction in synaptophysin immunoreactivity in the outer plexiform layer. The Muller cells were activated and expressed GFAP. No significant change of immunolabeling of MNF118 was found. These findings closely parallel those seen in human RP.

Animals↗
PubMed2002

Distribution and density of medium- and short-wavelength selective cones in the domestic pig retina.

The topography of medium (M)- and short (S)-wavelength sensitive cone photoreceptors was studied in the domestic pig retina. Antisera specific for M or S opsin as well as cone photoreceptor proteins arrestin and alpha-transducin were used to label cone types. Retinal wholemounts and their blood vessel patterns were drawn and specific regions removed. The wholemounts were immunocytochemically labelled to detect both M and S cones, and the specific regions labelled to detect S cones. Cones were counted in a 1 mm grid pattern, using the drawings as a guide. Pig retina has a high cone density retinal streak extending across the retina covering the optic disc (OD) and horizontal meridian. Densities in the streak are 20,000-35,000 mm(-2). Two higher peaks occur in the streak, one in temporal retina near the OD (39,000 mm(-2)) and the other in nasal retina 5-7 mm from the OD (40,500 mm(-2)). The lowest cone density is in far peripheral inferior retina (7000 mm(-2)). The total number of cones in pig retina is 17-20 million. Both types of cones are found throughout the retina, with S cone percentages ranging from 7.4 to 17.5% in no consistent topographical pattern. S cones have an irregular local distribution which can vary from a regular hexagonal pattern to small clusters of adjacent S cones to small areas lacking S cones. Double-label immunocytochemistry found that virtually all S cone outer segments (OS) contain some M opsin. M cone OS do not label at detectible levels for S opsin. Domestic pig retina is widely available, large, has a high cone density and has two types of cones. This tissue should be an excellent source for biochemical analysis of cone proteins, and for in vitro approaches to understanding cone survival factors.

Animals↗
PubMed2001

Photoreceptor subtype specification: from flies to humans.

Multiple cell types often differentiate from a pluripotent cell. These cells may then further diversify as distinct subtypes. The visual system provides an ideal model for studying subtype specification as various photoreceptors acquire different functions based on the type of opsin they express. Opsin expression is mostly controlled through transcriptional mechanisms that are evolutionary conserved from Drosophila to humans. In addition, it appears that, from a "default" developmental state, distinct "acquired" photoreceptor states develop upon receiving intrinsic or extrinsic signals. This review discusses factors involved in opsin gene regulation and how their integration may explain how subtype specificity is achieved.

Animals↗
PubMed1987

The pineal organ is the first differentiated light receptor in the embryonic salmon, Salmo salar L.

The initial appearance of S-antigen, alpha-transducin, opsin and 5-HT during embryogenesis of the pineal organ and retina was studied by means of immunocytochemistry in the Atlantic salmon, Salmo salar L. The presence of these substances may be taken as a good indication of photoreceptor differentiation; alpha-transducin and S-antigen are involved in the phototransduction process, opsin is the proteinaceous component of the photopigment rhodopsin, and 5-HT is a neurotransmitter or neurohormone produced by pineal photoreceptors. Two days after the retinal pigment layer became visible in the eggs, the outer segments of a few pineal photosensory cells showed immunoreactivity to opsin and alpha-transducin. At the same time S-antigen and serotonin were present in pineal cells of the photoreceptor type. The number of immunoreactive cells in the pineal organ increased up to hatching. In the differentiating retina of the salmon, no immunoreactivity to antibodies raised against the mentioned substances were detectable until after hatching. These results indicate that in ontogeny the developing pineal organ of the salmon embryo has the ability to perceive light information much earlier than the retina.

Animals↗
PubMed1987

Immunocytochemical localization of photopigments in cephalopod retinae.

The photopigments, rhodopsin and retinochrome, have been localized in cephalopod retinae using light and electron microscopic immunocytochemical methods. Polyclonal antibodies prepared against squid opsin demonstrated the presence of this protein in the photoreceptor rhabdomes, Golgi zone, Golgi-associated vesicles, plasma membrane, large cytoplasmic vesicles, and axonal membranes of octopus retinae. Monoclonal anti-opsin immunostained the rhabdomes and multivesicular bodies in the photoreceptor inner segments of squid. We believe the multivesicular bodies are involved in rhabdome turnover. Polyclonal anti-retinochrome localized this photopigment to the myeloid bodies of the photoreceptor inner segments, the rhabdomes, and to the extracellular space between opposing rhabdomeres in octopus retina. The results suggest some interesting functional relationships between rhodopsin and retinochrome with regard to chromophore exchange between illuminated forms of these photopigments and chromophore addition to newly synthesized opsin.

Animals↗
PubMed2007

Role of a novel photopigment, melanopsin, in behavioral adaptation to light.

Adaptation to changes in the ambient light is of critical importance to life. In mammals, three principal photoadaptation mechanisms depend on ocular photoreception and exhibit spectral sensitivity suggestive of the opsin class of photopigment(s). These include rapid adaptation of the visual system to the ambient light by pupil constriction, direct modulation of neuroendocrine function and entrainment of the circadian clock to the day:night cycle. Surprisingly, these processes can largely function independent of classical rod/cone photoreceptors, suggesting a novel opsin-based signaling mechanism. They appear to involve a recently discovered network of intrinsically photosensitive retinal ganglion cells that make direct or indirect axonal connections to brain centers regulating photoadaptive behaviors. The discovery of a novel opsin, melanopsin, in these cells has offered an exciting entry point to explore, at the molecular level, how mammals adapt to their light environment. There is now genetic proof of a principal role for melanopsin in all three major photoadaptation processes.

Adaptation, Biological↗
PubMed1998

Ciliary neurotrophic factor blocks rod photoreceptor differentiation from postmitotic precursor cells in vitro.

The development of photoreceptors in the mammalian retina is thought to be controlled by extrinsic signals. We have shown previously that ciliary neurotrophic factor (CNTF) potently inhibits photoreceptor differentiation in cultures of rat retina. The present study analyzes which developmental processes are affected by CNTF. Rod differentiation as determined by opsin and recoverin immunocytochemistry was effectively blocked by CNTF and leukemia inhibitory factor, but not by other neurotrophic agents tested. CNTF did not influence proliferation, cell death, or survival, and had no effect on the downregulation of nestin immunoreactivity in progenitor cells. Opsin-positive rods could be reverted to an opsin-negative state initially, but became unresponsive to CNTF later. No compensatory increase in the number of other cell types was observed. Application of neutralizing antibodies against CNTF revealed that rod development was partially blocked by an endogenous CNTF-like molecule in control cultures. Our results suggest that CNTF can act as a specific negative regulator of rod differentiation. Its action on photoreceptor precursor cells could serve to synchronize the maturation of photoreceptors, which are born over an extended period of time. Together with other stimulatory signals, CNTF may thus control the temporally and numerically correct integration of photoreceptors into the retinal network.

Animals↗
PubMed1988

Presence of retina-specific proteins in the lamprey pineal complex.

The pineal complex of river lamprey reacted with the antisera raised against retina specific proteins including bovine opsin, chick visinin and frog light-sensitive cyclic GMP phosphodiesterase (PDE). Immunoreactive materials stained with anti-opsin were evenly located at the outer segment of photoreceptor cells in the pineal organ and also found in the parapineal organ. Although anti-visinin stained the pineal and parapineal photoreceptor cells, the immunopositive photoreceptor cells were observed only at the lateral portion and not at the medial portion of the pineal organ. No immunoreactive materials were found in the pineal complex by the anti-PDE, whereas the anti-PDE reacted with photoreceptor cells of the retinal tissue. The data suggest that the pineal and parapineal retinas of lamprey contain opsin- and visinin-like proteins with different distribution in their photoreceptor cell layer as found in the lamprey retinal tissue.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1989

Alpha transducin is present in blue-, green-, and red-sensitive cone photoreceptors in the human retina.

Phototransduction in vertebrate rod and cone photoreceptor cells involves G protein-mediated light stimulation of cGMP hydrolysis. Enzymes of the cGMP hydrolysis cascades of rods and cones are products of different genes. Three different classes of cones in the human retina are maximally sensitive to either blue, green, or red light. Distinct opsin genes are expressed in each type of cone. The distribution of cone types in human retina was determined using anti-peptide antibodies that recognize specific amino acid sequences in green/red opsin and blue opsin. These antibodies together with an anti-peptide antibody against Tc alpha were used in double labeling experiments to demonstrate the presence of the Tc alpha peptide in all types of cones. cDNA clones corresponding to human rod and cone transducin alpha subunit (Tr alpha and Tc alpha) genes were isolated. Southern blot analyses of human genomic DNA suggest that there is only one rod T alpha gene but more than one cone T alpha gene. The multiple Tc alpha genes could be closely related genes or different Tc alpha alleles, or one could be a pseudogene.

Amino Acid Sequence↗
PubMed1988

Phenotypic expression of photoreceptor and endocrine cell properties by cultured pineal cells of the newborn rat.

Pineal glands of newborn rats were dissociated and maintained under cell culture conditions. The phenotypic expression of both photoreceptor and endocrine cell properties was investigated using immunohistochemical techniques (specific antibodies against opsin or serotonin). After one week in culture, a number of small round cells appeared on top of a sheet of flat epithelium. Among those cells, opsin-like immunoreactive cells were observed. These cells showed a neuron-like morphology with neuritic processes and often formed rosettes. Immunoreactivity was found on the plasma membrane of both the soma and cell processes. Serotonin-like immunoreactive cells were also differentiated in culture with two different morphological types of cells being found. One type resembled cultured serotonin-containing amacrine cells of the retina, and the other type had a flat, polygonal shape similar to that of pinealocytes. Both types of immunoreactive cells possessed fine neuritic processes. These results indicated that cell culture of rat pineal gland cells allowed expression of some properties, such as opsin synthesis and neuron-like morphology with long neuritic processes, that were not expressed in the intact rat pineal gland.

Animals↗
PubMed2005

Recombination reaction of rhodopsin in situ studied by photoconversion of "indicator yellow".

We measured the kinetics of recombination of 11-cis-retinal with opsin in intact frog rod outer segment (ROS). The rhodopsin in ROS was bleached and allowed to decay to "indicator yellow," a photoproduct where all-trans-retinal is partly free, and partly bound to non-specific amino groups of disk membranes. By briefly illuminating the "indicator yellow" by an intense 465 or 380-nm flash, we then photoconverted all-trans-retinal to (mostly) the 11-cis- form thus introducing into ROS a certain amount of cis-chromophore. The recombination of cis-retinal with opsin and the formation of rhodopsin were followed by fast single-cell microspectrophotometry. Regeneration proceeded with a time constant of approximately 3.5 min; up to 27% of bleached visual pigment was restored. The regenerated pigment consisted of 91% rhodopsin (11-cis-chromophore) and 9% of presumably isorhodopsin (9-cis-chromophore). The recombination of 11-cis-retinal with opsin inside the ROS proceeds substantially faster than rhodopsin regeneration in the intact eye and, hence, is not the rate-limiting step in the visual cycle.

Animals↗
PubMed1986

Chromophore/protein interaction in bacterial sensory rhodopsin and bacteriorhodopsin.

Retinal analogues with altered conjugated double bond systems or altered stereochemistry were incorporated into the phototaxis receptor sensory rhodopsin (SR) and the light-driven proton pump bacteriorhodopsin (BR) from Halobacterium halobium. Wavelength shifts in absorption ("opsin shifts") due to analogue interaction with the protein microenvironment demonstrate that the same overall electrostatic and steric properties of the retinal binding-site structures exist in both proteins despite their different functions. pi-Electron calculations from the opsin shifts lead to a new description of protein charge distribution that applies to the binding sites of both SR and BR. The new data extends the previously proposed external point charge model for BR to include an ion-pair protein/chromophore interaction near the beta-ionone moiety. The new data modifies the previously proposed external point-charge model, the derivation of which involved an experimentally erroneous opsin shift for one of the BR analogues.

Bacteriorhodopsins↗
PubMed2003

Evolutionary analysis of rhodopsin and cone pigments: connecting the three-dimensional structure with spectral tuning and signal transfer.

Extensive sequence data and structural sampling of expressed proteins from different species lead to the idea that entire molecules or specific domain folds belong to large superfamilies of proteins. A subset of G protein-coupled receptors, one of the largest families involved in cellular signaling, rod and cone opsins are involved in phototransduction in photoreceptor cells. Here, the evolutionary analysis of opsin sequences and structures predicts key residues involved in the transmission of the signal from the binding site of the chromophore to the cytoplasmic surface and residues that are involved in the spectral tuning of opsins to short wavelengths of light.

Amino Acid Sequence↗
PubMed1999

Evolution of visual pigments in geckos.

Most geckos are nocturnal forms and possess rod retinas, but some diurnal genera have pure-cone retinas. We isolated cDNAs encoding the diurnal gecko opsins, dg1 and dg2, similar to nocturnal gecko P521 and P467, respectively. Despite the large morphological differences between the diurnal and nocturnal gecko photoreceptor types, they express phylogenetically closely related opsins. These results provide molecular evidence for the reverse transmutation, that is, rods of an ancestral nocturnal gecko have backed into cones of diurnal geckos. The amino acid substitution rates of dgl and dg2 are higher than those of P521 and P467, respectively. Changes of behavior regarding photic environment may have contributed to acceleration of amino acid substitutions in the diurnal gecko opsins.

Amino Acid Sequence↗
PubMed2003

Multiple photopigments from the Mexican blind cavefish, Astyanax fasciatus: a microspectrophotometric study.

The cave-dwelling (hypogean) form of the teleost Astyanax fasciatus is blind, having only subdermal eye rudiments, but nevertheless maintains intact opsin genes. Second generation offspring of a cross between these and the normally sighted surface (epigean) form inherit opsin genes from both ancestries. A study of the expressed hypogean opsins of the hybrids, in comparison to the epigean forms, was undertaken by microspectrophotometry. The hybrid population showed considerable variation in the visual pigments of double cones, with evidence for two groups of cells with lambda(max) intermediate to those of the epigean pigments. Possible explanations for these intermediate pigments are discussed, including the hypothesis that they may represent hybrid genes similar to the genes for anomalous cone pigments in humans. Evidence was also found for ultraviolet-sensitive single cones and for an additional MWS pigment.

Animals↗