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Sample records for wave-sensitive lws opsins

  1. Genomic organization of duplicated short wave-sensitive and long wave-sensitive opsin genes in the green swordtail, Xiphophorus helleri

    PubMed Central

    2010-01-01

    Background Long wave-sensitive (LWS) opsin genes have undergone multiple lineage-specific duplication events throughout the evolution of teleost fishes. LWS repertoire expansions in live-bearing fishes (family Poeciliidae) have equipped multiple species in this family with up to four LWS genes. Given that color vision, especially attraction to orange male coloration, is important to mate choice within poeciliids, LWS opsins have been proposed as candidate genes driving sexual selection in this family. To date the genomic organization of these genes has not been described in the family Poeciliidae, and little is known about the mechanisms regulating the expression of LWS opsins in any teleost. Results Two BAC clones containing the complete genomic repertoire of LWS opsin genes in the green swordtail fish, Xiphophorus helleri, were identified and sequenced. Three of the four LWS loci identified here were linked in a tandem array downstream of two tightly linked short wave-sensitive 2 (SWS2) opsin genes. The fourth LWS opsin gene, containing only a single intron, was not linked to the other three and is the product of a retrotransposition event. Genomic and phylogenetic results demonstrate that the LWS genes described here share a common evolutionary origin with those previously characterized in other poeciliids. Using qualitative RT-PCR and MSP we showed that each of the LWS and SWS2 opsins, as well as three other cone opsin genes and a single rod opsin gene, were expressed in the eyes of adult female and male X. helleri, contributing to six separate classes of adult retinal cone and rod cells with average λmax values of 365 nm, 405 nm, 459 nm, 499 nm, 534 nm and 568 nm. Comparative genomic analysis identified two candidate teleost opsin regulatory regions containing putative CRX binding sites and hormone response elements in upstream sequences of LWS gene regions of seven teleost species, including X. helleri. Conclusions We report the first complete genomic description of LWS and SWS2 genes in poeciliids. These data will serve as a reference for future work seeking to understand the relationship between LWS opsin genomic organization, gene expression, gene family evolution, sexual selection and speciation in this fish family. PMID:20353595

  2. Androgens Increase lws Opsin Expression and Red Sensitivity in Male Three-Spined Sticklebacks

    PubMed Central

    Shao, Yi Ta; Wang, Feng-Yu; Fu, Wen-Chun; Anraku, Kazuhiko; Chen, I-Shiung

    2014-01-01

    Optomotor studies have shown that three-spined sticklebacks (Gasterosteus aculeatus) are more sensitive to red during summer than winter, which may be related to the need to detect the red breeding colour of males. This study aimed to determine whether this change of red light sensitivity is specifically related to reproductive physiology. The mRNA levels of opsin genes were examined in the retinae of sexually mature and immature fish, as well as in sham-operated males, castrated control males, or castrated males implanted with androgen 11-ketoandrostenedione (11 KA), maintained under stimulatory (L16:D8) or inhibitory (L8:D16) photoperiods. In both sexes, red-sensitive opsin gene (lws) mRNA levels were higher in sexually mature than in immature fish. Under L16:D8, lws mRNA levels were higher in intact than in castrated males, and were up-regulated by 11 KA treatment in castrated males. Moreover, electroretinogram data confirmed that sexual maturation resulted in higher relative red spectral sensitivity. Mature males under L16:D8 were more sensitive to red light than males under L8:D16. Red light sensitivity under L16:D8 was diminished by castration, but increased by 11 KA treatment. Thus, in sexually mature male sticklebacks, androgen is a key factor in enhancing sensitivity to red light via regulation of opsin gene expression. This is the first study to demonstrate that sex hormones can regulate spectral vision sensitivity. PMID:24963891

  3. Retinal photoreceptor arrangement, SWS1 and LWS opsin sequence, and electroretinography in the South American marsupial Thylamys elegans (Waterhouse, 1839).

    PubMed

    Palacios, Adrián G; Bozinovic, Francisco; Vielma, Alex; Arrese, Catherine A; Hunt, David M; Peichl, Leo

    2010-05-01

    We studied the retinal photoreceptors in the mouse opossum Thylamys elegans, a nocturnal South American marsupial. A variety of photoreceptor properties and color vision capabilities have been documented in Australian marsupials, and we were interested to establish what similarities and differences this American marsupial showed. Thylamys opsin gene sequencing revealed two cone opsins, a longwave-sensitive (LWS) opsin and a shortwave-sensitive (SWS1) opsin with deduced peak sensitivities at 560 nm and 360 nm (ultraviolet), respectively. Immunocytochemistry located these opsins to separate cone populations, a majority of LWS cones (density range 1,600-5,600/mm(2)) and a minority of SWS1 cones (density range 100-690/mm(2)). With rod densities of 440,000-590,000/mm(2), the cones constituted 0.4-1.2% of the photoreceptors. This is a suitable adaptation to nocturnal vision. Cone densities peaked in a horizontally elongated region ventral to the optic nerve head. In ventral-but not dorsal-retina, roughly 40% of the LWS opsin-expressing cones occurred as close pairs (double cones), and one member of each double cone contained a colorless oil droplet. The corneal electroretinogram (ERG) showed a high scotopic sensitivity with a rod peak sensitivity at 505 nm. At mesopic light levels, the spectral ERG revealed the contributions of a UV-sensitive SWS1 cone mechanism and an LWS cone mechanism with peak sensitivities at 365 nm and 555 nm, respectively, confirming the tuning predictions from the cone opsin sequences. The two spectral cone types provide the basis for dichromatic color vision, or trichromacy if the rods contribute to color processing at mesopic light levels. PMID:20187149

  4. Mix and match color vision: tuning spectral sensitivity by differential opsin gene expression in Lake Malawi cichlids.

    PubMed

    Parry, Juliet W L; Carleton, Karen L; Spady, Tyrone; Carboo, Aba; Hunt, David M; Bowmaker, James K

    2005-10-11

    Cichlid fish of the East African Rift Lakes are renowned for their diversity and offer a unique opportunity to study adaptive changes in the visual system in rapidly evolving species flocks. Since color plays a significant role in mate choice, differences in visual sensitivities could greatly influence and even drive speciation of cichlids. Lake Malawi cichlids inhabiting rock and sand habitats have significantly different cone spectral sensitivities. By combining microspectrophotometry (MSP) of isolated cones, sequencing of opsin genes, and spectral analysis of recombinant pigments, we have established the cone complements of four species of Malawi cichlids. MSP demonstrated that each of these species predominately expresses three cone pigments, although these differ between species to give three spectrally different cone complements. In addition, rare populations of spectrally distinct cones were found. In total, seven spectral classes were identified. This was confirmed by opsin gene sequencing, expression, and in vitro reconstitution. The genes represent the four major classes of cone opsin genes that diverged early in vertebrate evolution. All four species possess a long-wave-sensitive (LWS), three spectrally distinct green-sensitive (RH2), a blue-sensitive (SWS2A), a violet-sensitive (SWS2B), and an ultraviolet-sensitive (SWS1) opsin. However, African cichlids determine their spectral sensitivity by differential expression of primarily only three of the seven available cone opsin genes. Phylogenetic analysis suggests that all percomorph fish have similar potential. PMID:16213819

  5. Retinoic Acid Signaling Regulates Differential Expression of the Tandemly-Duplicated Long Wavelength-Sensitive Cone Opsin Genes in Zebrafish

    PubMed Central

    Frey, Ruth A.; Hunter, Samuel S.; Ashino, Ryuichi; Kawamura, Shoji; Stenkamp, Deborah L.

    2015-01-01

    The signaling molecule retinoic acid (RA) regulates rod and cone photoreceptor fate, differentiation, and survival. Here we elucidate the role of RA in differential regulation of the tandemly-duplicated long wavelength-sensitive (LWS) cone opsin genes. Zebrafish embryos were treated with RA from 48 hours post-fertilization (hpf) to 75 hpf, and RNA was isolated from eyes for microarray analysis. ~170 genes showed significantly altered expression, including several transcription factors and components of cellular signaling pathways. Of interest, the LWS1 opsin gene was strongly upregulated by RA. LWS1 is the upstream member of the tandemly duplicated LWS opsin array and is normally not expressed embryonically. Embryos treated with RA 48 hpf to 100 hpf or beyond showed significant reductions in LWS2-expressing cones in favor of LWS1-expressing cones. The LWS reporter line, LWS-PAC(H) provided evidence that individual LWS cones switched from LWS2 to LWS1 expression in response to RA. The RA signaling reporter line, RARE:YFP indicated that increased RA signaling in cones was associated with this opsin switch, and experimental reduction of RA signaling in larvae at the normal time of onset of LWS1 expression significantly inhibited LWS1 expression. A role for endogenous RA signaling in regulating differential expression of the LWS genes in postmitotic cones was further supported by the presence of an RA signaling domain in ventral retina of juvenile zebrafish that coincided with a ventral zone of LWS1 expression. This is the first evidence that an extracellular signal may regulate differential expression of opsin genes in a tandemly duplicated array. PMID:26296154

  6. Divergent selection for opsin gene variation in guppy (Poecilia reticulata) populations of Trinidad and Tobago

    PubMed Central

    Tezuka, A; Kasagi, S; van Oosterhout, C; McMullan, M; Iwasaki, W M; Kasai, D; Yamamichi, M; Innan, H; Kawamura, S; Kawata, M

    2014-01-01

    The guppy is known to exhibit remarkable interindividual variations in spectral sensitivity of middle to long wavelength-sensitive (M/LWS) cone photoreceptor cells. The guppy has four M/LWS-type opsin genes (LWS-1, LWS-2, LWS-3 and LWS-4) that are considered to be responsible for this sensory variation. However, the allelic variation of the opsin genes, particularly in terms of their absorption spectrum, has not been explored in wild populations. Thus, we examined nucleotide variations in the four M/LWS opsin genes as well as blue-sensitive SWS2-B and ultraviolet-sensitive SWS1 opsin genes for comparison and seven non-opsin nuclear loci as reference genes in 10 guppy populations from various light environments in Trinidad and Tobago. For the first time, we discovered a potential spectral variation (180 Ser/Ala) in LWS-1 that differed at an amino acid site known to affect the absorption spectra of opsins. Based on a coalescent simulation of the nucleotide variation of the reference genes, we showed that the interpopulation genetic differentiation of two opsin genes was significantly larger than the neutral expectation. Furthermore, this genetic differentiation was significantly related to differences in dissolved oxygen (DO) level, and it was not explained by the spatial distance between populations. The DO levels are correlated with eutrophication that possibly affects the color of aquatic environments. These results suggest that the population diversity of opsin genes is significantly driven by natural selection and that the guppy could adapt to various light environments through color vision changes. PMID:24690753

  7. Conservation, Duplication, and Divergence of Five Opsin Genes in Insect Evolution

    PubMed Central

    Feuda, Roberto; Marlétaz, Ferdinand; Bentley, Michael A.; Holland, Peter W.H.

    2016-01-01

    Opsin proteins covalently bind to small molecular chromophores and each protein-chromophore complex is sensitive to particular wavelengths of light. Multiple opsins with different wavelength absorbance peaks are required for color vision. Comparing opsin responses is challenging at low light levels, explaining why color vision is often lost in nocturnal species. Here, we investigated opsin evolution in 27 phylogenetically diverse insect species including several transitions between photic niches (nocturnal, diurnal, and crepuscular). We find widespread conservation of five distinct opsin genes, more than commonly considered. These comprise one c-opsin plus four r-opsins (long wavelength sensitive or LWS, blue sensitive, ultra violet [UV] sensitive and the often overlooked Rh7 gene). Several recent opsin gene duplications are also detected. The diversity of opsin genes is consistent with color vision in diurnal, crepuscular, and nocturnal insects. Tests for positive selection in relation to photic niche reveal evidence for adaptive evolution in UV-sensitive opsins in day-flying insects in general, and in LWS opsins of day-flying Lepidoptera specifically. PMID:26865071

  8. Conservation, Duplication, and Divergence of Five Opsin Genes in Insect Evolution.

    PubMed

    Feuda, Roberto; Marlétaz, Ferdinand; Bentley, Michael A; Holland, Peter W H

    2016-01-01

    Opsin proteins covalently bind to small molecular chromophores and each protein-chromophore complex is sensitive to particular wavelengths of light. Multiple opsins with different wavelength absorbance peaks are required for color vision. Comparing opsin responses is challenging at low light levels, explaining why color vision is often lost in nocturnal species. Here, we investigated opsin evolution in 27 phylogenetically diverse insect species including several transitions between photic niches (nocturnal, diurnal, and crepuscular). We find widespread conservation of five distinct opsin genes, more than commonly considered. These comprise one c-opsin plus four r-opsins (long wavelength sensitive or LWS, blue sensitive, ultra violet [UV] sensitive and the often overlooked Rh7 gene). Several recent opsin gene duplications are also detected. The diversity of opsin genes is consistent with color vision in diurnal, crepuscular, and nocturnal insects. Tests for positive selection in relation to photic niche reveal evidence for adaptive evolution in UV-sensitive opsins in day-flying insects in general, and in LWS opsins of day-flying Lepidoptera specifically. PMID:26865071

  9. Anion sensitivity and spectral tuning of middle- and long-wavelength-sensitive (MWS/LWS) visual pigments.

    PubMed

    Davies, Wayne I L; Wilkie, Susan E; Cowing, Jill A; Hankins, Mark W; Hunt, David M

    2012-07-01

    The long-wavelength-sensitive (LWS) opsins form one of four classes of vertebrate cone visual pigment and exhibit peak spectral sensitivities (λ(max)) that generally range from 525 to 560 nm for rhodopsin/vitamin-A(1) photopigments. Unique amongst the opsin classes, many LWS pigments show anion sensitivity through the interaction of chloride ions with a histidine residue at site 197 (H197) to give a long-wavelength spectral shift in peak sensitivity. Although it has been shown that amino acid substitutions at five sites (180, 197, 277, 285 and 308) are useful in predicting the λ(max) values of the LWS pigment class, some species, such as the elephant shark and most marine mammals, express LWS opsins that possess λ(max) values that are not consistent with this 'five-site' rule, indicating that other interactions may be involved. This study has taken advantage of the natural mutation at the chloride-binding site in the mouse LWS pigment. Through the use of a number of mutant pigments generated by site-directed mutagenesis, a new model has been formulated that takes into account the role of charge and steric properties of the side chains of residues at sites 197 and 308 in the function of the chloride-binding site in determining the peak sensitivity of LWS photopigments. PMID:22349213

  10. Molecular cloning and expression of long-wavelength-sensitive cone opsin in the brain of a tropical damselfish.

    PubMed

    Takeuchi, Yuki; Bapary, Mohammad Abu Jafor; Igarashi, Syugo; Imamura, Satoshi; Sawada, Yuji; Matsumoto, Mio; Hur, Sung-Pyo; Takemura, Akihiro

    2011-12-01

    Ovarian development of the sapphire devil, Chrysiptera cyanea, exhibits photoperiodism and is stimulated under long-day conditions. Previous studies suggest that red light is more effective than green and blue lights for inducing ovarian development. In addition, the extra-retinal photoreception involved in the ovarian development is suggested in this species. The present study aimed to clone the red-light-sensitive cone opsin (sdLWS) of this species, to demonstrate its expression in the brain, and to confirm the effectiveness of red light on the initiation of ovarian development. A phylogenetic analysis revealed that sdLWS belongs to the long-wavelength-sensitive opsin (LWS opsin) group, showing high identity (77-92%) with the LWS opsin of other fishes. RT-PCR showed that sdLWS is expressed in the eye, brain, and ovary. In situ hybridization indicated that sdLWS is expressed in the third ventricle periventricular area in the anterior hypothalamus. Exposing fish to long-day conditions of red light resulted in the appearance of vitellogenic oocytes in the ovary and an increase in the gonadosomatic index within 2weeks, suggesting that fish immediately undergo oocyte development under red light conditions. These results indicate that sdLWS is a possible candidate of deep brain photoreceptor molecule involved in photoperiodic ovarian development in the sapphire devil. PMID:21871576

  11. Rod monochromacy and the coevolution of cetacean retinal opsins.

    PubMed

    Meredith, Robert W; Gatesy, John; Emerling, Christopher A; York, Vincent M; Springer, Mark S

    2013-04-01

    Cetaceans have a long history of commitment to a fully aquatic lifestyle that extends back to the Eocene. Extant species have evolved a spectacular array of adaptations in conjunction with their deployment into a diverse array of aquatic habitats. Sensory systems are among those that have experienced radical transformations in the evolutionary history of this clade. In the case of vision, previous studies have demonstrated important changes in the genes encoding rod opsin (RH1), short-wavelength sensitive opsin 1 (SWS1), and long-wavelength sensitive opsin (LWS) in selected cetaceans, but have not examined the full complement of opsin genes across the complete range of cetacean families. Here, we report protein-coding sequences for RH1 and both color opsin genes (SWS1, LWS) from representatives of all extant cetacean families. We examine competing hypotheses pertaining to the timing of blue shifts in RH1 relative to SWS1 inactivation in the early history of Cetacea, and we test the hypothesis that some cetaceans are rod monochomats. Molecular evolutionary analyses contradict the "coastal" hypothesis, wherein SWS1 was pseudogenized in the common ancestor of Cetacea, and instead suggest that RH1 was blue-shifted in the common ancestor of Cetacea before SWS1 was independently knocked out in baleen whales (Mysticeti) and in toothed whales (Odontoceti). Further, molecular evidence implies that LWS was inactivated convergently on at least five occasions in Cetacea: (1) Balaenidae (bowhead and right whales), (2) Balaenopteroidea (rorquals plus gray whale), (3) Mesoplodon bidens (Sowerby's beaked whale), (4) Physeter macrocephalus (giant sperm whale), and (5) Kogia breviceps (pygmy sperm whale). All of these cetaceans are known to dive to depths of at least 100 m where the underwater light field is dim and dominated by blue light. The knockout of both SWS1 and LWS in multiple cetacean lineages renders these taxa rod monochromats, a condition previously unknown among mammalian species. PMID:23637615

  12. Rod Monochromacy and the Coevolution of Cetacean Retinal Opsins

    PubMed Central

    Meredith, Robert W.; Gatesy, John; Emerling, Christopher A.; York, Vincent M.; Springer, Mark S.

    2013-01-01

    Cetaceans have a long history of commitment to a fully aquatic lifestyle that extends back to the Eocene. Extant species have evolved a spectacular array of adaptations in conjunction with their deployment into a diverse array of aquatic habitats. Sensory systems are among those that have experienced radical transformations in the evolutionary history of this clade. In the case of vision, previous studies have demonstrated important changes in the genes encoding rod opsin (RH1), short-wavelength sensitive opsin 1 (SWS1), and long-wavelength sensitive opsin (LWS) in selected cetaceans, but have not examined the full complement of opsin genes across the complete range of cetacean families. Here, we report protein-coding sequences for RH1 and both color opsin genes (SWS1, LWS) from representatives of all extant cetacean families. We examine competing hypotheses pertaining to the timing of blue shifts in RH1 relative to SWS1 inactivation in the early history of Cetacea, and we test the hypothesis that some cetaceans are rod monochomats. Molecular evolutionary analyses contradict the “coastal” hypothesis, wherein SWS1 was pseudogenized in the common ancestor of Cetacea, and instead suggest that RH1 was blue-shifted in the common ancestor of Cetacea before SWS1 was independently knocked out in baleen whales (Mysticeti) and in toothed whales (Odontoceti). Further, molecular evidence implies that LWS was inactivated convergently on at least five occasions in Cetacea: (1) Balaenidae (bowhead and right whales), (2) Balaenopteroidea (rorquals plus gray whale), (3) Mesoplodon bidens (Sowerby's beaked whale), (4) Physeter macrocephalus (giant sperm whale), and (5) Kogia breviceps (pygmy sperm whale). All of these cetaceans are known to dive to depths of at least 100 m where the underwater light field is dim and dominated by blue light. The knockout of both SWS1 and LWS in multiple cetacean lineages renders these taxa rod monochromats, a condition previously unknown among mammalian species. PMID:23637615

  13. Short Wavelength Cone Opsin Is Not Expressed in the Retina of Arboreal African Pangolin (Manis tricuspis)

    PubMed Central

    Adekanmbi, Adejoke J.; Adekanmbi, Adefisayo A.; Akinola, Oluwole B.

    2016-01-01

    This paper reports a study of cone photoreceptors present in the retina of Manis tricuspis. Specifically, the LWS (L-) opsin expressed in longwave-sensitive cones and SWS1 (S-) opsin shortwave-sensitive cones were targeted. Vertical sections revealed reactivity to a cone marker, peanut agglutinin (PNA), and to an LWS antibody, but not to an SWS1 antibody. This suggests that the Manis tricuspis visual system is not able to discriminate shorter wavelengths from longer wavelengths because the short wavelength cones are not expressed in their retina.

  14. Evolution of opsin expression in birds driven by sexual selection and habitat.

    PubMed

    Bloch, Natasha I

    2015-01-01

    Theories of sexual and natural selection predict coevolution of visual perception with conspecific colour and/or the light environment animals occupy. One way to test these theories is to focus on the visual system, which can be achieved by studying the opsin-based visual pigments that mediate vision. Birds vary greatly in colour, but opsin gene coding sequences and associated visual pigment spectral sensitivities are known to be rather invariant across birds. Here, I studied expression of the four cone opsin genes (Lws, Rh2, Sws2 and Sws1) in 16 species of New World warblers (Parulidae). I found levels of opsin expression vary both across species and between the sexes. Across species, female, but not male Sws2 expression is associated with an index of sexual selection, plumage dichromatism. This fits predictions of classic sexual selection models, in which the sensory system changes in females, presumably impacting female preference, and co-evolves with male plumage. Expression of the opsins at the extremes of the light spectrum, Lws and Uvs, correlates with the inferred light environment occupied by the different species. Unlike opsin spectral tuning, regulation of opsin gene expression allows for fast adaptive evolution of the visual system in response to natural and sexual selection, and in particular, sex-specific selection pressures. PMID:25429020

  15. Evolution of opsin expression in birds driven by sexual selection and habitat

    PubMed Central

    Bloch, Natasha I.

    2015-01-01

    Theories of sexual and natural selection predict coevolution of visual perception with conspecific colour and/or the light environment animals occupy. One way to test these theories is to focus on the visual system, which can be achieved by studying the opsin-based visual pigments that mediate vision. Birds vary greatly in colour, but opsin gene coding sequences and associated visual pigment spectral sensitivities are known to be rather invariant across birds. Here, I studied expression of the four cone opsin genes (Lws, Rh2, Sws2 and Sws1) in 16 species of New World warblers (Parulidae). I found levels of opsin expression vary both across species and between the sexes. Across species, female, but not male Sws2 expression is associated with an index of sexual selection, plumage dichromatism. This fits predictions of classic sexual selection models, in which the sensory system changes in females, presumably impacting female preference, and co-evolves with male plumage. Expression of the opsins at the extremes of the light spectrum, Lws and Uvs, correlates with the inferred light environment occupied by the different species. Unlike opsin spectral tuning, regulation of opsin gene expression allows for fast adaptive evolution of the visual system in response to natural and sexual selection, and in particular, sex-specific selection pressures. PMID:25429020

  16. RT-qPCR reveals opsin gene upregulation associated with age and sex in guppies (Poecilia reticulata) - a species with color-based sexual selection and 11 visual-opsin genes

    PubMed Central

    2011-01-01

    Background PCR-based surveys have shown that guppies (Poecilia reticulata) have an unusually large visual-opsin gene repertoire. This has led to speculation that opsin duplication and divergence has enhanced the evolution of elaborate male coloration because it improves spectral sensitivity and/or discrimination in females. However, this conjecture on evolutionary connections between opsin repertoire, vision, mate choice, and male coloration was generated with little data on gene expression. Here, we used RT-qPCR to survey visual-opsin gene expression in the eyes of males, females, and juveniles in order to further understand color-based sexual selection from the perspective of the visual system. Results Juvenile and adult (male and female) guppies express 10 visual opsins at varying levels in the eye. Two opsin genes in juveniles, SWS2B and RH2-2, accounted for >85% of all visual-opsin transcripts in the eye, excluding RH1. This relative abundance (RA) value dropped to about 65% in adults, as LWS-A180 expression increased from approximately 3% to 20% RA. The juvenile-to-female transition also showed LWS-S180 upregulation from about 1.5% to 7% RA. Finally, we found that expression in guppies' SWS2-LWS gene cluster is negatively correlated with distance from a candidate locus control region (LCR). Conclusions Selective pressures influencing visual-opsin gene expression appear to differ among age and sex. LWS upregulation in females is implicated in augmenting spectral discrimination of male coloration and courtship displays. In males, enhanced discrimination of carotenoid-rich food and possibly rival males are strong candidate selective pressures driving LWS upregulation. These developmental changes in expression suggest that adults possess better wavelength discrimination than juveniles. Opsin expression within the SWS2-LWS gene cluster appears to be regulated, in part, by a common LCR. Finally, by comparing our RT-qPCR data to MSP data, we were able to propose the first opsin-to-λmax assignments for all photoreceptor types in the cone mosaic. PMID:21447186

  17. Euarchontan Opsin Variation Brings New Focus to Primate Origins.

    PubMed

    Melin, Amanda D; Wells, Konstans; Moritz, Gillian L; Kistler, Logan; Orkin, Joseph D; Timm, Robert M; Bernard, Henry; Lakim, Maklarin B; Perry, George H; Kawamura, Shoji; Dominy, Nathaniel J

    2016-04-01

    Debate on the adaptive origins of primates has long focused on the functional ecology of the primate visual system. For example, it is hypothesized that variable expression of short- (SWS1) and middle-to-long-wavelength sensitive (M/LWS) opsins, which confer color vision, can be used to infer ancestral activity patterns and therefore selective ecological pressures. A problem with this approach is that opsin gene variation is incompletely known in the grandorder Euarchonta, that is, the orders Scandentia (treeshrews), Dermoptera (colugos), and Primates. The ancestral state of primate color vision is therefore uncertain. Here, we report on the genes (OPN1SW and OPN1LW) that encode SWS1 and M/LWS opsins in seven species of treeshrew, including the sole nocturnal scandentian Ptilocercus lowii. In addition, we examined the opsin genes of the Central American woolly opossum (Caluromys derbianus), an enduring ecological analogue in the debate on primate origins. Our results indicate: 1) retention of ultraviolet (UV) visual sensitivity in C. derbianus and a shift from UV to blue spectral sensitivities at the base of Euarchonta; 2) ancient pseudogenization of OPN1SW in the ancestors of P. lowii, but a signature of purifying selection in those of C. derbianus; and, 3) the absence of OPN1LW polymorphism among diurnal treeshrews. These findings suggest functional variation in the color vision of nocturnal mammals and a distinctive visual ecology of early primates, perhaps one that demanded greater spatial resolution under light levels that could support cone-mediated color discrimination. PMID:26739880

  18. Euarchontan Opsin Variation Brings New Focus to Primate Origins

    PubMed Central

    Melin, Amanda D.; Wells, Konstans; Moritz, Gillian L.; Kistler, Logan; Orkin, Joseph D.; Timm, Robert M.; Bernard, Henry; Lakim, Maklarin B.; Perry, George H.; Kawamura, Shoji; Dominy, Nathaniel J.

    2016-01-01

    Debate on the adaptive origins of primates has long focused on the functional ecology of the primate visual system. For example, it is hypothesized that variable expression of short- (SWS1) and middle-to-long-wavelength sensitive (M/LWS) opsins, which confer color vision, can be used to infer ancestral activity patterns and therefore selective ecological pressures. A problem with this approach is that opsin gene variation is incompletely known in the grandorder Euarchonta, that is, the orders Scandentia (treeshrews), Dermoptera (colugos), and Primates. The ancestral state of primate color vision is therefore uncertain. Here, we report on the genes (OPN1SW and OPN1LW) that encode SWS1 and M/LWS opsins in seven species of treeshrew, including the sole nocturnal scandentian Ptilocercus lowii. In addition, we examined the opsin genes of the Central American woolly opossum (Caluromys derbianus), an enduring ecological analogue in the debate on primate origins. Our results indicate: 1) retention of ultraviolet (UV) visual sensitivity in C. derbianus and a shift from UV to blue spectral sensitivities at the base of Euarchonta; 2) ancient pseudogenization of OPN1SW in the ancestors of P. lowii, but a signature of purifying selection in those of C. derbianus; and, 3) the absence of OPN1LW polymorphism among diurnal treeshrews. These findings suggest functional variation in the color vision of nocturnal mammals and a distinctive visual ecology of early primates, perhaps one that demanded greater spatial resolution under light levels that could support cone-mediated color discrimination. PMID:26739880

  19. Short- and long-wavelength-sensitive opsins are involved in photoreception both in the retina and throughout the central nervous system of crayfish.

    PubMed

    Kingston, Alexandra C N; Cronin, Thomas W

    2015-12-01

    Crayfish have two classes of photoreceptors in the retinas of their reflecting superposition eyes. Long-wavelength-sensitive photoreceptors, comprised of microvilli from R1-7 cells, make up the main rhabdoms. Eighth retinular cells, located distal to the main rhabdoms, house short-wavelength-sensitive photoreceptors. While the opsin involved in long-wavelength sensitivity has long been known, we present the first description of the short-wavelength-sensitive opsin in the retina of the red swamp crayfish, Procambarus clarkii. The expression patterns of these SWS and LWS opsin proteins in the retina are consistent with the previously described locations of SWS and LWS receptors. Crayfish also have a well-characterized extraocular photoreceptor, called the caudal photoreceptor, located in the sixth abdominal ganglion. To search for retinal opsins in the caudal photoreceptor (and elsewhere in the CNS), we used RT-PCR and immunohistochemical labeling. We found both SWS and LWS opsin transcripts not only in the sixth abdominal ganglion, but also in all ganglia of the nerve cord. Immunolabeling shows that both opsins are expressed in nerve fibers that extend from the brain through the entire length of the CNS. Thus, the same two photopigments are used both for vision in the retina and for extraocular functions throughout the CNS of crayfish. PMID:26445969

  20. Opsin evolution in the Ambulacraria.

    PubMed

    D'Aniello, S; Delroisse, J; Valero-Gracia, A; Lowe, E K; Byrne, M; Cannon, J T; Halanych, K M; Elphick, M R; Mallefet, J; Kaul-Strehlow, S; Lowe, C J; Flammang, P; Ullrich-Lüter, E; Wanninger, A; Arnone, M I

    2015-12-01

    Opsins--G-protein coupled receptors involved in photoreception--have been extensively studied in the animal kingdom. The present work provides new insights into opsin-based photoreception and photoreceptor cell evolution with a first analysis of opsin sequence data for a major deuterostome clade, the Ambulacraria. Systematic data analysis, including for the first time hemichordate opsin sequences and an expanded echinoderm dataset, led to a robust opsin phylogeny for this cornerstone superphylum. Multiple genomic and transcriptomic resources were surveyed to cover each class of Hemichordata and Echinodermata. In total, 119 ambulacrarian opsin sequences were found, 22 new sequences in hemichordates and 97 in echinoderms (including 67 new sequences). We framed the ambulacrarian opsin repertoire within eumetazoan diversity by including selected reference opsins from non-ambulacrarians. Our findings corroborate the presence of all major ancestral bilaterian opsin groups in Ambulacraria. Furthermore, we identified two opsin groups specific to echinoderms. In conclusion, a molecular phylogenetic framework for investigating light-perception and photobiological behaviors in marine deuterostomes has been obtained. PMID:26472700

  1. Molecular and functional characterization of opsins in barfin flounder (Verasper moseri).

    PubMed

    Kasagi, Satoshi; Mizusawa, Kanta; Murakami, Naoto; Andoh, Tadashi; Furufuji, Sumihisa; Kawamura, Shoji; Takahashi, Akiyoshi

    2015-02-10

    Green light irradiation facilitates the somatic growth of barfin flounder (Verasper moseri). However, the V. moseri visual system, which may be associated with somatic growth by acting on the endocrine system upon exposure to this particular wavelength, remains largely unexplored. Herein, we characterized the visual opsin repertoire of V. moseri to understand the molecular basis underlying this effect. The five types of visual opsins that are found in vertebrates were cloned from RNA that was extracted from the eyes of V. moseri. Notably, V. moseri possessed one pseudogene (RH2-A) and two intact (RH2-B and RH2-C) copies of "green-sensitive" opsin genes. The wavelengths of maximum absorption spectra (λmax) for each of the reconstituted photopigments were 552nm for "red-sensitive" LWS, 506nm for RH2-B, 490nm for RH2-C, 482nm and 416nm for "blue-sensitive" SWS2A and SWS2B, respectively, 367nm for "ultraviolet-sensitive" SWS1, and 494nm for "dim-light sensitive rhodopsin" RH1. The λmax of SWS2A was longer than that of any other reported vertebrate SWS2 opsin. By measuring the expression level of these opsin genes with quantitative RT-PCR in 3-, 15-, and 27-month-old fish, we found that RH2-B and SWS2A were expressed at a constant level, whereas the expression of LWS, RH2-C, SWS2B, and SWS1 opsin genes decreased, and that of RH1 increased with age. Barfin flounders inhabit inshore waters at a young age and expand their habitat to deep sea areas as they age, and green light is relatively abundant in deep water compared to the lights of other wavelengths in shallow water. Our results indicate that gene repertoire and expression profile of the opsin genes of barfin flounder are adaptive to their habitat shift that occurs during development, with some opsins acquiring a distinct λmax. PMID:25433330

  2. Evolution and functional diversity of jellyfish opsins.

    PubMed

    Suga, Hiroshi; Schmid, Volker; Gehring, Walter J

    2008-01-01

    Cnidaria are the most basal animal phylum possessing complex eyes [1]. Their eyes predominantly use ciliary photoreceptor cells (c-PRCs) like vertebrates, whereas insect eyes use rhabdomeric photoreceptor cells (r-PRCs) [1-4]. These two cell types show not only different cytoarchitectures but distinct phototransduction cascades, which are triggered by the respective types of opsins (e.g., [5]), ciliary opsins (c-opsins) and rhabdomeric opsins (r-opsins) [6]. Recent reports suggested that the c- and r-PRCs and their respective opsins diverged at least before the deuterostome-protostome split [7-9]. To study the earlier evolution of animal PRCs and opsins, we investigated two hydrozoan jellyfishes. We report here the first-characterized cnidarian opsins. Molecular phylogeny revealed that the cloned 20 jellyfish opsins, together with all the opsins from a hydra and some from a sea anemone, are more closely related to the c-opsins than to any other major opsin subfamily, indicating that the divergence of c- and r-opsins antedates the Cnidaria-Bilateria split. Possible scenarios of animal PRC evolution are discussed. Furthermore, Cladonema opsins show several distinct tissue- and stage-specific expression patterns. The expression of specific opsins in the eyes suggests a role in vision, whereas that in the gonads suggests a role in light-controlled release of gametes. PMID:18160295

  3. Variable light environments induce plastic spectral tuning by regional opsin coexpression in the African cichlid fish, Metriaclima zebra.

    PubMed

    Dalton, Brian E; Lu, Jessica; Leips, Jeff; Cronin, Thomas W; Carleton, Karen L

    2015-08-01

    Critical behaviours such as predation and mate choice often depend on vision. Visual systems are sensitive to the spectrum of light in their environment, which can vary extensively both within and among habitats. Evolutionary changes in spectral sensitivity contribute to divergence and speciation. Spectral sensitivity of the retina is primarily determined by visual pigments, which are opsin proteins bound to a chromophore. We recently discovered that photoreceptors in different regions of the retina, which view objects against distinct environmental backgrounds, coexpress different pairs of opsins in an African cichlid fish, Metriaclima zebra. This coexpression tunes the sensitivity of the retinal regions to the corresponding backgrounds and may aid in detection of dark objects, such as predators. Although intraretinal regionalization of spectral sensitivity in many animals correlates with their light environments, it is unknown whether variation in the light environment induces developmentally plastic alterations of intraretinal sensitivity regions. Here, we demonstrate with fluorescent in situ hybridization and qPCR that the spectrum and angle of environmental light both influence the development of spectral sensitivity regions by altering the distribution and level of opsins across the retina. Normally, M. zebra coexpresses LWS opsin with RH2Aα opsin in double cones of the ventral but not the dorsal retina. However, when illuminated from below throughout development, adult M. zebra coexpressed LWS and RH2Aα in double cones both dorsally and ventrally. Thus, environmental background spectra alter the spectral sensitivity pattern that develops across the retina, potentially influencing behaviours and related evolutionary processes such as courtship and speciation. PMID:26175094

  4. LWS/SET Technology Experiment Carrier

    NASA Technical Reports Server (NTRS)

    Sherman, Barry; Giffin, Geoff

    2002-01-01

    This paper examines the approach taken to building a low-cost, modular spacecraft bus that can be used to support a variety of technology experiments in different space environments. It describes the techniques used and design drivers considered to ensure experiment independence from as yet selected host spacecraft. It describes the technology experiment carriers that will support NASA's Living With a Star Space Environment Testbed space missions. NASA has initiated the Living With a Star (LWS) Program to develop a better scientific understanding to address the aspects of the connected Sun-Earth system that affect life and society. A principal goal of the program is to bridge the gap between science, engineering, and user application communities. The Space Environment Testbed (SET) Project is one element of LWS. The Project will enable future science, operational, and commercial objectives in space and atmospheric environments by improving engineering approaches to the accommodation and/or mitigation of the effects of solar variability on technological systems. The SET Project is highly budget constrained and must seek to take advantage of as yet undetermined partnering opportunities for access to space. SET will conduct technology validation experiments hosted on available flight opportunities. The SET Testbeds will be developed in a manner that minimizes the requirements for accommodation, and will be flown as flight opportunities become available. To access the widest range of flight opportunities, two key development requirements are to maintain flexibility with respect to accommodation constraints and to have the capability to respond quickly to flight opportunities. Experiments, already developed to the technology readiness level of needing flight validation in the variable Sun-Earth environment, will be selected on the basis of the need for the subject technology, readiness for flight, need for flight resources and particular orbit. Experiments will be accumulated by the Project and manifested for specific flight opportunities as they become available. The SET Carrier is designed to present a standard set of interfaces to SET technology experiments and to be modular and flexible enough to interface to a variety of possible host spacecraft. The Carrier will have core components and mission unique components. Once the core carrier elements have been developed, only the mission unique components need to be defined and developed for any particular mission. This approach will minimize the mission specific cost and development schedule for a given flight opportunity. The standard set of interfaces provided by SET to experiments allows them to be developed independent of the particulars of a host spacecraft. The Carrier will provide the power, communication, and the necessary monitoring features to operate experiments. The Carrier will also provide all of the mechanical assemblies and harnesses required to adapt experiments to a particular host. Experiments may be hosted locally with the Carrier or remotely on the host spacecraft. The Carrier design will allow a single Carrier to support a variable number of experiments and will include features that support the ability to incrementally add experiments without disturbing the core architecture.

  5. Intraspecific cone opsin expression variation in the cichlids of Lake Malawi.

    PubMed

    Smith, Adam R; D'Annunzio, Lindsay; Smith, Abbi E; Sharma, Anit; Hofmann, Christopher M; Marshall, N J; Carleton, Karen L

    2011-01-01

    The expression of cone opsin genes is a primary determinant of the characteristics of colour vision. Interspecific variation in opsin expression is common in African cichlids. It is correlated with foraging among cichlids from Lake Malawi, and with ambient light environment among cichlids from Lake Victoria. In this study, we tested whether gene expression varied within species such that it might be important in contributing to divergence. We hypothesized that light attenuation with depth would be correlated with predictable changes in gene expression in Lake Malawi, and that this variation would tune visual sensitivities to match the ambient light environment. We observed significant differences in cone opsin expression in three different comparisons among populations of the same species. Higher LWS expression was found in shallow versus deep Copadichromis eucinostomus. In Metriaclima zebra, individuals from Zimbawe Rock expressed significantly more SWS2B than those from Thumbi West Island, although these locales have similar ambient light environments. Finally, Tropheops gracilior from deeper water had significantly more variation in expression than their shallow counterparts. These results support that gene expression varies significantly between populations of the same species. Surprisingly, these results could not be explained by predicted visual performance as models predicted that differential expression patterns did not confer sensitivity advantages at different depths. This suggested that expression variation did not confer a local sensitivity advantage. Therefore, our findings were contrary to a primary requirement of the sensory bias hypothesis. As such, other explanations for intraspecific gene expression variation need to be tested. PMID:21091561

  6. Vision for the Future of Lws TR&T

    NASA Astrophysics Data System (ADS)

    Schwadron, N.; Mannucci, A. J.; Antiochos, S. K.; Bhattacharjee, A.; Gombosi, T. I.; Gopalswamy, N.; Kamalabadi, F.; Linker, J.; Pilewskie, P.; Pulkkinen, A. A.; Spence, H. E.; Tobiska, W. K.; Weimer, D. R.; Withers, P.; Bisi, M. M.; Kuznetsova, M. M.; Miller, K. L.; Moretto, T.; Onsager, T. G.; Roussev, I. I.; Viereck, R. A.

    2014-12-01

    The Living With a Star (LWS) program addresses acute societal needs for understanding the effects of space weather and developing scientific knowledge to support predictive capabilities. Our society's heavy reliance on technologies affected by the space environment, an enormous number of airline customers, interest in space tourism, and the developing plans for long-duration human exploration space missions are clear examples that demonstrate urgent needs for space weather models and detailed understanding of space weather effects and risks. Since its inception, the LWS program has provided a vehicle to innovate new mechanisms for conducting research, building highly effective interdisciplinary teams, and ultimately in developing the scientific understanding needed to transition research tools into operational models that support the predictive needs of our increasingly space-reliant society. The advances needed require broad-based observations that cannot be obtained by large missions alone. The Decadal Survey (HDS, 2012) outlines the nation's needs for scientific development that will build the foundation for tomorrow's space weather services. Addressing these goals, LWS must develop flexible pathways to space utilizing smaller, more diverse and rapid development of observational platforms. Expanding utilization of ground-based assets and shared launches will also significantly enhance opportunities to fulfill the growing LWS data needs. Partnerships between NASA divisions, national/international agencies, and with industry will be essential for leveraging resources to address increasing societal demand for space weather advances. Strengthened connections to user communities will enhance the quality and impact of deliverables from LWS programs. Thus, we outline the developing vision for the future of LWS, stressing the need for deeper scientific understanding to improve forecasting capabilities, for more diverse data resources, and for project deliverables that address the growing needs of user communities.

  7. Opsin vs opsin: New materials for biotechnological applications

    SciTech Connect

    Alfinito, Eleonora; Reggiani, Lino

    2014-08-14

    The need of new diagnostic methods satisfying, as an early detection, a low invasive procedure and a cost-efficient value, is orienting the technological research toward the use of bio-integrated devices, in particular, bio-sensors. The set of know-why necessary to achieve this goal is wide, from biochemistry to electronics and is summarized in an emerging branch of electronics, called proteotronics. Proteotronics is here applied to state a comparative analysis of the electrical responses coming from type-1 and type-2 opsins. In particular, the procedure is used as an early investigation of a recently discovered family of opsins, the proteorhodopsins activated by blue light, BPRs. The results reveal some interesting and unexpected similarities between proteins of the two families, suggesting the global electrical response are not strictly linked to the class identity.

  8. Chromatic organization of cone photoreceptors in the retina of rainbow trout: single cones irreversibly switch from UV (SWS1) to blue (SWS2) light sensitive opsin during natural development.

    PubMed

    Cheng, Christiana L; Flamarique, Iñigo Novales

    2007-12-01

    The retinas of salmonid fishes have single and double cones arranged in square to row formations termed mosaics. The square mosaic unit is formed by four double cones that make the sides of the square with a single (centre) cone in the middle, and a single (corner) cone at each corner of the square when present. Previous research using coho salmon-derived riboprobes on four species of anadromous Pacific salmon has shown that all single cones express a SWS1 (UV sensitive) visual pigment protein (opsin) at hatching, and that these cones switch to a SWS2 (blue light sensitive) opsin during the juvenile period. Whether this opsin switch applies to non-anadromous species, like the rainbow trout, is under debate as species-specific riboprobes have not been used to study opsin expression during development of a trout. As well, a postulated recovery of SWS1 opsin expression in the retina of adult rainbow trout, perhaps via a reverse process to that occurring in the juvenile, has not been investigated. Here, we used in situ hybridization with species-specific riboprobes and microspectrophotometry on rainbow trout retina to show that: (1) single cones in the juvenile switch opsin expression from SWS1 to SWS2, (2) this switch is not reversed in the adult, i.e. all single cones in the main retina continue to express SWS2 opsin, and (3) opsin switches do not occur in double cones: each member expresses one opsin, maximally sensitive to green (RH2) or red (LWS) light. The opsin switch in the single cones of salmonid fishes may be a general process of chromatic organization that occurs during retinal development of most vertebrates. PMID:18025012

  9. Diversity of Active States in TMT Opsins

    PubMed Central

    Sakai, Kazumi; Yamashita, Takahiro; Imamoto, Yasushi; Shichida, Yoshinori

    2015-01-01

    Opn3/TMT opsins belong to one of the opsin groups with vertebrate visual and non-visual opsins, and are widely distributed in eyes, brains and other internal organs in various vertebrates and invertebrates. Vertebrate Opn3/TMT opsins are further classified into four groups on the basis of their amino acid identities. However, there is limited information about molecular properties of these groups, due to the difficulty in preparing the recombinant proteins. Here, we successfully expressed recombinant proteins of TMT1 and TMT2 opsins of medaka fish (Oryzias latipes) in cultured cells and characterized their molecular properties. Spectroscopic and biochemical studies demonstrated that TMT1 and TMT2 opsins functioned as blue light-sensitive Gi/Go-coupled receptors, but exhibited spectral properties and photo-convertibility of the active state different from each other. TMT1 opsin forms a visible light-absorbing active state containing all-trans-retinal, which can be photo-converted to 7-cis- and 9-cis-retinal states in addition to the original 11-cis-retinal state. In contrast, the active state of TMT2 opsin is a UV light-absorbing state having all-trans-retinal and does not photo-convert to any other state, including the original 11-cis-retinal state. Thus, TMT opsins are diversified so as to form a different type of active state, which may be responsible for their different functions. PMID:26491964

  10. Shedding new light on opsin evolution

    PubMed Central

    Porter, Megan L.; Blasic, Joseph R.; Bok, Michael J.; Cameron, Evan G.; Pringle, Thomas; Cronin, Thomas W.; Robinson, Phyllis R.

    2012-01-01

    Opsin proteins are essential molecules in mediating the ability of animals to detect and use light for diverse biological functions. Therefore, understanding the evolutionary history of opsins is key to understanding the evolution of light detection and photoreception in animals. As genomic data have appeared and rapidly expanded in quantity, it has become possible to analyse opsins that functionally and histologically are less well characterized, and thus to examine opsin evolution strictly from a genetic perspective. We have incorporated these new data into a large-scale, genome-based analysis of opsin evolution. We use an extensive phylogeny of currently known opsin sequence diversity as a foundation for examining the evolutionary distributions of key functional features within the opsin clade. This new analysis illustrates the lability of opsin protein-expression patterns, site-specific functionality (i.e. counterion position) and G-protein binding interactions. Further, it demonstrates the limitations of current model organisms, and highlights the need for further characterization of many of the opsin sequence groups with unknown function. PMID:22012981

  11. Analysis of the Opsin Repertoire in the Tardigrade Hypsibius dujardini Provides Insights into the Evolution of Opsin Genes in Panarthropoda

    PubMed Central

    Hering, Lars; Mayer, Georg

    2014-01-01

    Screening of a deeply sequenced transcriptome using Illumina sequencing as well as the genome of the tardigrade Hypsibius dujardini revealed a set of five opsin genes. To clarify the phylogenetic position of these genes and to elucidate the evolutionary history of opsins in Panarthropoda (Onychophora + Tardigrada + Arthropoda), we reconstructed the phylogeny of broadly sampled metazoan opsin genes using maximum likelihood and Bayesian inference methods in conjunction with carefully selected substitution models. According to our findings, the opsin repertoire of H. dujardini comprises representatives of all three major bilaterian opsin clades, including one r-opsin, three c-opsins, and a Group 4 opsin (neuropsin/opsin-5). The identification of the tardigrade ortholog of neuropsin/opsin-5 is the first record of this opsin type in a protostome, but our screening of available metazoan genomes revealed that it is also present in other protostomes. Our opsin phylogeny further suggests that two r-opsins, including an “arthropsin,” were present in the last common ancestor of Panarthropoda. Although both r-opsin lineages were retained in Onychophora and Arthropoda, the arthropsin was lost in Tardigrada. The single (most likely visual) r-opsin found in H. dujardini supports the hypothesis of monochromatic vision in the panarthropod ancestor, whereas two duplications of the ancestral panarthropod c-opsin have led to three c-opsins in tardigrades. Although the early-branching nodes are unstable within the metazoans, our findings suggest that the last common ancestor of Bilateria possessed six opsins: Two r-opsins, one c-opsin, and three Group 4 opsins, one of which (Go opsin) was lost in the ecdysozoan lineage. PMID:25193307

  12. Analysis of the opsin repertoire in the tardigrade Hypsibius dujardini provides insights into the evolution of opsin genes in panarthropoda.

    PubMed

    Hering, Lars; Mayer, Georg

    2014-09-01

    Screening of a deeply sequenced transcriptome using Illumina sequencing as well as the genome of the tardigrade Hypsibius dujardini revealed a set of five opsin genes. To clarify the phylogenetic position of these genes and to elucidate the evolutionary history of opsins in Panarthropoda (Onychophora + Tardigrada + Arthropoda), we reconstructed the phylogeny of broadly sampled metazoan opsin genes using maximum likelihood and Bayesian inference methods in conjunction with carefully selected substitution models. According to our findings, the opsin repertoire of H. dujardini comprises representatives of all three major bilaterian opsin clades, including one r-opsin, three c-opsins, and a Group 4 opsin (neuropsin/opsin-5). The identification of the tardigrade ortholog of neuropsin/opsin-5 is the first record of this opsin type in a protostome, but our screening of available metazoan genomes revealed that it is also present in other protostomes. Our opsin phylogeny further suggests that two r-opsins, including an "arthropsin," were present in the last common ancestor of Panarthropoda. Although both r-opsin lineages were retained in Onychophora and Arthropoda, the arthropsin was lost in Tardigrada. The single (most likely visual) r-opsin found in H. dujardini supports the hypothesis of monochromatic vision in the panarthropod ancestor, whereas two duplications of the ancestral panarthropod c-opsin have led to three c-opsins in tardigrades. Although the early-branching nodes are unstable within the metazoans, our findings suggest that the last common ancestor of Bilateria possessed six opsins: Two r-opsins, one c-opsin, and three Group 4 opsins, one of which (Go opsin) was lost in the ecdysozoan lineage. PMID:25193307

  13. The Microbial Opsin Family of Optogenetic Tools

    PubMed Central

    Zhang, Feng; Vierock, Johannes; Yizhar, Ofer; Fenno, Lief E.; Tsunoda, Satoshi; Kianianmomeni, Arash; Prigge, Matthias; Berndt, Andre; Cushman, John; Polle, Jürgen; Magnuson, Jon; Hegemann, Peter; Deisseroth, Karl

    2014-01-01

    The capture and utilization of light is an exquisitely evolved process. The single-component microbial opsins, although more limited than multicomponent cascades in processing, display unparalleled compactness and speed. Recent advances in understanding microbial opsins have been driven by molecular engineering for optogenetics and by comparative genomics. Here we provide a Primer on these light-activated ion channels and pumps, describe a group of opsins bridging prior categories, and explore the convergence of molecular engineering and genomic discovery for the utilization and understanding of these remarkable molecular machines. PMID:22196724

  14. The Microbial Opsin Family of Optogenetic Tools

    SciTech Connect

    Zhang, Feng; Vierock, Johannes; Yizhar, Ofer; Fenno, Lief E.; Tsunoda, Satoshi; Kianianmomeni, Arash; Prigge, Matthias; Berndt, Andre; Cushman, John C.; Polle, Juergen E.; Magnuson, Jon K.; Hegemann, Peter; Deisseroth, Karl

    2011-12-23

    The capture and utilization of light is an exquisitely evolved process. The single-component microbial opsins, although more limited than multicomponent cascades in processing, display unparalleled compactness and speed. Recent advances in understanding microbial opsins have been driven by molecular engineering for optogenetics and by comparative genomics. Here we provide a Primer on these light-activated ion channels and pumps, describe a group of opsins bridging prior categories, and explore the convergence of molecular engineering and genomic discovery for the utilization and understanding of these remarkable molecular machines.

  15. Involvement of opsins in mammalian sperm thermotaxis

    PubMed Central

    Pérez-Cerezales, Serafín; Boryshpolets, Sergii; Afanzar, Oshri; Brandis, Alexander; Nevo, Reinat; Kiss, Vladimir; Eisenbach, Michael

    2015-01-01

    A unique characteristic of mammalian sperm thermotaxis is extreme temperature sensitivity, manifested by the capacity of spermatozoa to respond to temperature changes of <0.0006 °C as they swim their body-length distance. The identity of the sensing system that confers this exceptional sensitivity on spermatozoa is not known. Here we show that the temperature-sensing system of mammalian spermatozoa involves opsins, known to be G-protein-coupled receptors that act as photosensors in vision. We demonstrate by molecular, immunological, and functional approaches that opsins are present in human and mouse spermatozoa at specific sites, which depend on the species and the opsin type, and that they are involved in sperm thermotaxis via two signalling pathways—the phospholipase C and the cyclic-nucleotide pathways. Our results suggest that, depending on the context and the tissue, mammalian opsins act not only as photosensors but also as thermosensors. PMID:26537127

  16. Broad-Band Activatable White-Opsin

    PubMed Central

    Batabyal, Subrata; Cervenka, Gregory; Ha, Ji Hee; Kim, Young-tae; Mohanty, Samarendra

    2015-01-01

    Currently, the use of optogenetic sensitization of retinal cells combined with activation/inhibition has the potential to be an alternative to retinal implants that would require electrodes inside every single neuron for high visual resolution. However, clinical translation of optogenetic activation for restoration of vision suffers from the drawback that the narrow spectral sensitivity of an opsin requires active stimulation by a blue laser or a light emitting diode with much higher intensities than ambient light. In order to allow an ambient light-based stimulation paradigm, we report the development of a ‘white-opsin’ that has broad spectral excitability in the visible spectrum. The cells sensitized with white-opsin showed excitability at an order of magnitude higher with white light compared to using only narrow-band light components. Further, cells sensitized with white-opsin produced a photocurrent that was five times higher than Channelrhodopsin-2 under similar photo-excitation conditions. The use of fast white-opsin may allow opsin-sensitized neurons in a degenerated retina to exhibit a higher sensitivity to ambient white light. This property, therefore, significantly lowers the activation threshold in contrast to conventional approaches that use intense narrow-band opsins and light to activate cellular stimulation. PMID:26360377

  17. Metazoan opsin evolution reveals a simple route to animal vision

    PubMed Central

    Feuda, Roberto; Hamilton, Sinead C.; McInerney, James O.; Pisani, Davide

    2012-01-01

    All known visual pigments in Neuralia (Cnidaria, Ctenophora, and Bilateria) are composed of an opsin (a seven-transmembrane G protein-coupled receptor), and a light-sensitive chromophore, generally retinal. Accordingly, opsins play a key role in vision. There is no agreement on the relationships of the neuralian opsin subfamilies, and clarifying their phylogeny is key to elucidating the origin of this protein family and of vision. We used improved methods and data to resolve the opsin phylogeny and explain the evolution of animal vision. We found that the Placozoa have opsins, and that the opsins share a common ancestor with the melatonin receptors. Further to this, we found that all known neuralian opsins can be classified into the same three subfamilies into which the bilaterian opsins are classified: the ciliary (C), rhabdomeric (R), and go-coupled plus retinochrome, retinal G protein-coupled receptor (Go/RGR) opsins. Our results entail a simple scenario of opsin evolution. The first opsin originated from the duplication of the common ancestor of the melatonin and opsin genes in a eumetazoan (Placozoa plus Neuralia) ancestor, and an inference of its amino acid sequence suggests that this protein might not have been light-sensitive. Two more gene duplications in the ancestral neuralian lineage resulted in the origin of the R, C, and Go/RGR opsins. Accordingly, the first animal with at least a C, an R, and a Go/RGR opsin was a neuralian progenitor. PMID:23112152

  18. Metazoan opsin evolution reveals a simple route to animal vision.

    PubMed

    Feuda, Roberto; Hamilton, Sinead C; McInerney, James O; Pisani, Davide

    2012-11-13

    All known visual pigments in Neuralia (Cnidaria, Ctenophora, and Bilateria) are composed of an opsin (a seven-transmembrane G protein-coupled receptor), and a light-sensitive chromophore, generally retinal. Accordingly, opsins play a key role in vision. There is no agreement on the relationships of the neuralian opsin subfamilies, and clarifying their phylogeny is key to elucidating the origin of this protein family and of vision. We used improved methods and data to resolve the opsin phylogeny and explain the evolution of animal vision. We found that the Placozoa have opsins, and that the opsins share a common ancestor with the melatonin receptors. Further to this, we found that all known neuralian opsins can be classified into the same three subfamilies into which the bilaterian opsins are classified: the ciliary (C), rhabdomeric (R), and go-coupled plus retinochrome, retinal G protein-coupled receptor (Go/RGR) opsins. Our results entail a simple scenario of opsin evolution. The first opsin originated from the duplication of the common ancestor of the melatonin and opsin genes in a eumetazoan (Placozoa plus Neuralia) ancestor, and an inference of its amino acid sequence suggests that this protein might not have been light-sensitive. Two more gene duplications in the ancestral neuralian lineage resulted in the origin of the R, C, and Go/RGR opsins. Accordingly, the first animal with at least a C, an R, and a Go/RGR opsin was a neuralian progenitor. PMID:23112152

  19. The Living With a Star (LWS) Sentinels Mission

    NASA Technical Reports Server (NTRS)

    Szabo, A.

    2005-01-01

    The Sentinels Mission, the heliospheric element of the NASA Living With a Star (LWS) program, is still rapidly evolving, especially as the Sentinels Science and Technology Definition Team is progressing with its work. With the Solar Dynamics Observatory, the solar component, and the Geospace elements taking a more finalized form, it becomes clearer what scientific and measurement objectives will be necessary to establish the solar-geospace connection in order to achieve the goals of the LWS program. Possible, early formulation designs of the Sentinels mission will be presented that includes the Inner Heliospheric Mappers, a four spacecraft mission to observe the inner heliosphere between 0.25 and 1.0 AUs along with a Far Side Sentinel that will perform remote solar observations from nearly the opposite side of the Sun. Moreover, the complementarity of the various planned international missions (e.g., ESA Solar Orbiter, and Beppi Colombo) along with NASA planetary projects (e.g., Mars program and MESSENGER) will be discussed and how they can form a coherent system. Finally, the importance of already available heliospheric data will be emphasized.

  20. The vertebrate ancestral repertoire of visual opsins, transducin alpha subunits and oxytocin/vasopressin receptors was established by duplication of their shared genomic region in the two rounds of early vertebrate genome duplications

    PubMed Central

    2013-01-01

    Background Vertebrate color vision is dependent on four major color opsin subtypes: RH2 (green opsin), SWS1 (ultraviolet opsin), SWS2 (blue opsin), and LWS (red opsin). Together with the dim-light receptor rhodopsin (RH1), these form the family of vertebrate visual opsins. Vertebrate genomes contain many multi-membered gene families that can largely be explained by the two rounds of whole genome duplication (WGD) in the vertebrate ancestor (2R) followed by a third round in the teleost ancestor (3R). Related chromosome regions resulting from WGD or block duplications are said to form a paralogon. We describe here a paralogon containing the genes for visual opsins, the G-protein alpha subunit families for transducin (GNAT) and adenylyl cyclase inhibition (GNAI), the oxytocin and vasopressin receptors (OT/VP-R), and the L-type voltage-gated calcium channels (CACNA1-L). Results Sequence-based phylogenies and analyses of conserved synteny show that the above-mentioned gene families, and many neighboring gene families, expanded in the early vertebrate WGDs. This allows us to deduce the following evolutionary scenario: The vertebrate ancestor had a chromosome containing the genes for two visual opsins, one GNAT, one GNAI, two OT/VP-Rs and one CACNA1-L gene. This chromosome was quadrupled in 2R. Subsequent gene losses resulted in a set of five visual opsin genes, three GNAT and GNAI genes, six OT/VP-R genes and four CACNA1-L genes. These regions were duplicated again in 3R resulting in additional teleost genes for some of the families. Major chromosomal rearrangements have taken place in the teleost genomes. By comparison with the corresponding chromosomal regions in the spotted gar, which diverged prior to 3R, we could time these rearrangements to post-3R. Conclusions We present an extensive analysis of the paralogon housing the visual opsin, GNAT and GNAI, OT/VP-R, and CACNA1-L gene families. The combined data imply that the early vertebrate WGD events contributed to the evolution of vision and the other neuronal and neuroendocrine functions exerted by the proteins encoded by these gene families. In pouched lamprey all five visual opsin genes have previously been identified, suggesting that lampreys diverged from the jawed vertebrates after 2R. PMID:24180662

  1. The comb jelly opsins and the origins of animal phototransduction.

    PubMed

    Feuda, Roberto; Rota-Stabelli, Omar; Oakley, Todd H; Pisani, Davide

    2014-08-01

    Opsins mediate light detection in most animals, and understanding their evolution is key to clarify the origin of vision. Despite the public availability of a substantial collection of well-characterized opsins, early opsin evolution has yet to be fully understood, in large part because of the high level of divergence observed among opsins belonging to different subfamilies. As a result, different studies have investigated deep opsin evolution using alternative data sets and reached contradictory results. Here, we integrated the data and methods of three, key, recent studies to further clarify opsin evolution. We show that the opsin relationships are sensitive to outgroup choice; we generate new support for the existence of Rhabdomeric opsins in Cnidaria (e.g., corals and jellyfishes) and show that all comb jelly opsins belong to well-recognized opsin groups (the Go-coupled opsins or the Ciliary opsins), which are also known in Bilateria (e.g., humans, fruit flies, snails, and their allies) and Cnidaria. Our results are most parsimoniously interpreted assuming a traditional animal phylogeny where Ctenophora are not the sister group of all the other animals. PMID:25062921

  2. The Comb Jelly Opsins and the Origins of Animal Phototransduction

    PubMed Central

    Feuda, Roberto; Rota-Stabelli, Omar; Oakley, Todd H.; Pisani, Davide

    2014-01-01

    Opsins mediate light detection in most animals, and understanding their evolution is key to clarify the origin of vision. Despite the public availability of a substantial collection of well-characterized opsins, early opsin evolution has yet to be fully understood, in large part because of the high level of divergence observed among opsins belonging to different subfamilies. As a result, different studies have investigated deep opsin evolution using alternative data sets and reached contradictory results. Here, we integrated the data and methods of three, key, recent studies to further clarify opsin evolution. We show that the opsin relationships are sensitive to outgroup choice; we generate new support for the existence of Rhabdomeric opsins in Cnidaria (e.g., corals and jellyfishes) and show that all comb jelly opsins belong to well-recognized opsin groups (the Go-coupled opsins or the Ciliary opsins), which are also known in Bilateria (e.g., humans, fruit flies, snails, and their allies) and Cnidaria. Our results are most parsimoniously interpreted assuming a traditional animal phylogeny where Ctenophora are not the sister group of all the other animals. PMID:25062921

  3. Losses of functional opsin genes, short-wavelength cone photopigments, and color vision--a significant trend in the evolution of mammalian vision.

    PubMed

    Jacobs, Gerald H

    2013-03-01

    All mammalian cone photopigments are derived from the operation of representatives from two opsin gene families (SWS1 and LWS in marsupial and eutherian mammals; SWS2 and LWS in monotremes), a process that produces cone pigments with respective peak sensitivities in the short and middle-to-long wavelengths. With the exception of a number of primate taxa, the modal pattern for mammals is to have two types of cone photopigment, one drawn from each of the gene families. In recent years, it has been discovered that the SWS1 opsin genes of a widely divergent collection of eutherian mammals have accumulated mutational changes that render them nonfunctional. This alteration reduces the retinal complements of these species to a single cone type, thus rendering ordinary color vision impossible. At present, several dozen species from five mammalian orders have been identified as falling into this category, but the total number of mammalian species that have lost short-wavelength cones in this way is certain to be much larger, perhaps reaching as high as 10% of all species. A number of circumstances that might be used to explain this widespread cone loss can be identified. Among these, the single consistent fact is that the species so affected are nocturnal or, if they are not technically nocturnal, they at least feature retinal organizations that are typically associated with that lifestyle. At the same time, however, there are many nocturnal mammals that retain functional short-wavelength cones. Nocturnality thus appears to set the stage for loss of functional SWS1 opsin genes in mammals, but it cannot be the sole circumstance. PMID:23286388

  4. Cubozoan genome illuminates functional diversification of opsins and photoreceptor evolution.

    PubMed

    Liegertová, Michaela; Pergner, Jiří; Kozmiková, Iryna; Fabian, Peter; Pombinho, Antonio R; Strnad, Hynek; Pačes, Jan; Vlček, Čestmír; Bartůněk, Petr; Kozmik, Zbyněk

    2015-01-01

    Animals sense light primarily by an opsin-based photopigment present in a photoreceptor cell. Cnidaria are arguably the most basal phylum containing a well-developed visual system. The evolutionary history of opsins in the animal kingdom has not yet been resolved. Here, we study the evolution of animal opsins by genome-wide analysis of the cubozoan jellyfish Tripedalia cystophora, a cnidarian possessing complex lens-containing eyes and minor photoreceptors. A large number of opsin genes with distinct tissue- and stage-specific expression were identified. Our phylogenetic analysis unequivocally classifies cubozoan opsins as a sister group to c-opsins and documents lineage-specific expansion of the opsin gene repertoire in the cubozoan genome. Functional analyses provided evidence for the use of the Gs-cAMP signaling pathway in a small set of cubozoan opsins, indicating the possibility that the majority of other cubozoan opsins signal via distinct pathways. Additionally, these tests uncovered subtle differences among individual opsins, suggesting possible fine-tuning for specific photoreceptor tasks. Based on phylogenetic, expression and biochemical analysis we propose that rapid lineage- and species-specific duplications of the intron-less opsin genes and their subsequent functional diversification promoted evolution of a large repertoire of both visual and extraocular photoreceptors in cubozoans. PMID:26154478

  5. A mammalian neural tissue opsin (Opsin 5) is a deep brain photoreceptor in birds

    PubMed Central

    Nakane, Yusuke; Ikegami, Keisuke; Ono, Hiroko; Yamamoto, Naoyuki; Yoshida, Shosei; Hirunagi, Kanjun; Ebihara, Shizufumi; Kubo, Yoshihiro; Yoshimura, Takashi

    2010-01-01

    It has been known for many decades that nonmammalian vertebrates detect light by deep brain photoreceptors that lie outside the retina and pineal organ to regulate seasonal cycle of reproduction. However, the identity of these photoreceptors has so far remained unclear. Here we report that Opsin 5 is a deep brain photoreceptive molecule in the quail brain. Expression analysis of members of the opsin superfamily identified as Opsin 5 (OPN5; also known as Gpr136, Neuropsin, PGR12, and TMEM13) mRNA in the paraventricular organ (PVO), an area long believed to be capable of phototransduction. Immunohistochemistry identified Opsin 5 in neurons that contact the cerebrospinal fluid in the PVO, as well as fibers extending to the external zone of the median eminence adjacent to the pars tuberalis of the pituitary gland, which translates photoperiodic information into neuroendocrine responses. Heterologous expression of Opsin 5 in Xenopus oocytes resulted in light-dependent activation of membrane currents, the action spectrum of which showed peak sensitivity (λmax) at ∼420 nm. We also found that short-wavelength light, i.e., between UV-B and blue light, induced photoperiodic responses in eye-patched, pinealectomized quail. Thus, Opsin 5 appears to be one of the deep brain photoreceptive molecules that regulates seasonal reproduction in birds. PMID:20679218

  6. Multiple rod–cone and cone–rod photoreceptor transmutations in snakes: Evidence from visual opsin gene expression

    USGS Publications Warehouse

    Simoe, Bruno F; Sampaio, Filipa L.; Loew, Ellis R.; Sanders, Kate L.; Fisher, Robert N.; Hart, Nathan S.; Hunt, David M.; Partridge, Julian C.; Gower, David J.

    2016-01-01

    In 1934, Gordon Walls forwarded his radical theory of retinal photoreceptor ‘transmutation’. This proposed that rods and cones used for scotopic and photopic vision, respectively, were not fixed but could evolve into each other via a series of morphologically distinguishable intermediates. Walls' prime evidence came from series of diurnal and nocturnal geckos and snakes that appeared to have pure-cone or pure-rod retinas (in forms that Walls believed evolved from ancestors with the reverse complement) or which possessed intermediate photoreceptor cells. Walls was limited in testing his theory because the precise identity of visual pigments present in photoreceptors was then unknown. Subsequent molecular research has hitherto neglected this topic but presents new opportunities. We identify three visual opsin genes, rh1, sws1 and lws, in retinal mRNA of an ecologically and taxonomically diverse sample of snakes central to Walls' theory. We conclude that photoreceptors with superficially rod- or cone-like morphology are not limited to containing scotopic or photopic opsins, respectively. Walls' theory is essentially correct, and more research is needed to identify the patterns, processes and functional implications of transmutation. Future research will help to clarify the fundamental properties and physiology of photoreceptors adapted to function in different light levels.

  7. Multiple rod-cone and cone-rod photoreceptor transmutations in snakes: evidence from visual opsin gene expression.

    PubMed

    Simões, Bruno F; Sampaio, Filipa L; Loew, Ellis R; Sanders, Kate L; Fisher, Robert N; Hart, Nathan S; Hunt, David M; Partridge, Julian C; Gower, David J

    2016-01-27

    In 1934, Gordon Walls forwarded his radical theory of retinal photoreceptor 'transmutation'. This proposed that rods and cones used for scotopic and photopic vision, respectively, were not fixed but could evolve into each other via a series of morphologically distinguishable intermediates. Walls' prime evidence came from series of diurnal and nocturnal geckos and snakes that appeared to have pure-cone or pure-rod retinas (in forms that Walls believed evolved from ancestors with the reverse complement) or which possessed intermediate photoreceptor cells. Walls was limited in testing his theory because the precise identity of visual pigments present in photoreceptors was then unknown. Subsequent molecular research has hitherto neglected this topic but presents new opportunities. We identify three visual opsin genes, rh1, sws1 and lws, in retinal mRNA of an ecologically and taxonomically diverse sample of snakes central to Walls' theory. We conclude that photoreceptors with superficially rod- or cone-like morphology are not limited to containing scotopic or photopic opsins, respectively. Walls' theory is essentially correct, and more research is needed to identify the patterns, processes and functional implications of transmutation. Future research will help to clarify the fundamental properties and physiology of photoreceptors adapted to function in different light levels. PMID:26817768

  8. Carrier Plus: A Sensor Payload for Living With a Star Space Environment Testbed (LWS/SET)

    NASA Technical Reports Server (NTRS)

    Marshall, Cheryl; Moss, Steven; Howard, Regan; LaBel, Kenneth; Grycewicz, Tom; Barth, Janet; Brewer, Dana

    2003-01-01

    The paper discusses the following: 1. Living with a Star (LWS) program: space environment testbed (SET); natural space environment. 2. Carrier plus: goals and benefits. 3. ON-orbit sensor measurements. 4. Carrier plus architecture. 5. Participation in carrier plus.

  9. The Dynamic Evolutionary History of Pancrustacean Eyes and Opsins.

    PubMed

    Henze, Miriam J; Oakley, Todd H

    2015-11-01

    Pancrustacea (Hexapoda plus Crustacea) display an enormous diversity of eye designs, including multiple types of compound eyes and single-chambered eyes, often with color vision and/or polarization vision. Although the eyes of some pancrustaceans are well-studied, there is still much to learn about the evolutionary paths to this amazing visual diversity. Here, we examine the evolutionary history of eyes and opsins across the principle groups of Pancrustacea. First, we review the distribution of lateral and median eyes, which are found in all major pancrustacean clades (Oligostraca, Multicrustacea, and Allotriocarida). At the same time, each of those three clades has taxa that lack lateral and/or median eyes. We then compile data on the expression of visual r-opsins (rhabdomeric opsins) in lateral and median eyes across Pancrustacea and find no evidence for ancient opsin clades expressed in only one type of eye. Instead, opsin clades with eye-specific expression are products of recent gene duplications, indicating a dynamic past, during which opsins often changed expression from one type of eye to another. We also investigate the evolutionary history of peropsins and r-opsins, which are both known to be expressed in eyes of arthropods. By searching published transcriptomes, we discover for the first time crustacean peropsins and suggest that previously reported odonate opsins may also be peropsins. Finally, from analyzing a reconciled, phylogenetic tree of arthropod r-opsins, we infer that the ancestral pancrustacean had four visual opsin genes, which we call LW2, MW1, MW2, and SW. These are the progenitors of opsin clades that later were variously duplicated or lost during pancrustacean evolution. Together, our results reveal a particularly dynamic history, with losses of eyes, duplication and loss of opsin genes, and changes in opsin expression between types of eyes. PMID:26319405

  10. ISO LWS observations of planetary nebula fine-structure lines

    NASA Astrophysics Data System (ADS)

    Liu, X.-W.; Barlow, M. J.; Cohen, M.; Danziger, I. J.; Luo, S.-G.; Baluteau, J. P.; Cox, P.; Emery, R. J.; Lim, T.; Péquignot, D.

    2001-05-01

    We have obtained 43-198μm far-infrared (IR) spectra for a sample of 51 Galactic planetary nebulae (PN) and protoplanetary nebulae (PPN), using the Long Wavelength Spectrometer (LWS) on board the Infrared Space Observatory (ISO). Spectra were also obtained of the former PN candidate Lo 14. The spectra yield fluxes for the fine-structure lines [Nii] 122μm, [Niii] 57μm and [Oiii] 52 and 88μm emitted in the ionized regions and the [Oi] 63- and 146-μm and [Cii] 158-μm lines from the photodissociation regions (PDRs), which have been used to determine electron densities and ionic abundances for the ionized regions and densities, temperatures and gas masses for the PDRs. The strong [Niii] and [Oiii] emission lines detected in the LWS spectrum taken centred on Lo 14 could be associated with the nearby strong radio and infrared source G 331.5-0.1. We find that the electron densities yielded by the [Oiii] 88μm/52μm doublet ratio are systematically lower than those derived from the optical [Ariv] λ4740/λ4711 and [Cliii] λ5537/λ5517 doublet ratios, which have much higher critical densities than the 52- and 88-μm lines, suggesting the presence of density inhomogeneities in the nebulae. Ionic abundances, N+/H+, N2+/H+ and O2+/H+, as well as the N2+/O2+ abundance ratio, which provides a good approximation to the N/O elemental abundance ratio, are derived. Although ionic abundances relative to H+ deduced from the far-IR fine-structure lines are sensitive to the adopted electron density and the presence of density inhomogeneities, the strong dependence on the nebular physical conditions is largely cancelled out when N2+/O2+ is calculated from the 57μm/(52μm+88μm) flux ratio, owing to the similarity of the critical densities of the lines involved. The temperatures and densities of the PDRs around 24 PN have been determined from the observed [Oi] and [Cii] line intensity ratios. Except for a few objects, the deduced temperatures fall between 200 and 500K, peaking around 250K. The densities of the PDRs vary from 104-105cm-3, reaching 3×105cm-3 in some young compact PN. With a derived temperature of 1600K and a density of 105cm-3, the PDR of NGC 7027 is one of the warmest and at the same time one of the densest amongst the nebulae studied. For most of the PN studied, the [Cii]-emitting regions contain only modest amounts of material, with gas masses <~0.1Msolar. Exceptional large PDR masses are found for a few nebulae, including NGC 7027, the bipolar nebulae M2-9 and NGC 6302, the young dense planetary nebulae BD+30°3639, IC 418 and NGC 5315, and the old, probably recombining, nebulae IC 4406 and NGC 6072.

  11. Carrier Plus: A sensor payload for Living With a Star Space Environment Testbed (LWS/SET)

    NASA Technical Reports Server (NTRS)

    Marshall, Cheryl J.; Moss, Steven; Howard, Regan; LaBel, Kenneth A.; Grycewicz, Tom; Barth, Janet L.; Brewer, Dana

    2003-01-01

    The Defense Threat Reduction Agency (DTR4) and National Aeronautics and Space Administration (NASA) Goddard Space Flight Center are collaborating to develop the Carrier Plus sensor experiment platform as a capability of the Space Environments Testbed (SET). The Space Environment Testbed (SET) provides flight opportunities for technology experiments as part of NASA's Living With a Star (LWS) program. The Carrier Plus will provide new capability to characterize sensor technologies such as state-of-the-art visible focal plane arrays (FPAs) in a natural space radiation environment. The technical objectives include on-orbit validation of recently developed FPA technologies and performance prediction methodologies, as well as characterization of the FPA radiation response to total ionizing dose damage, displacement damage and transients. It is expected that the sensor experiment will carry 4-6 FPAs and associated radiation correlative environment monitors (CEMs) for a 2006-2007 launch. Sensor technology candidates may include n- and p-charge coupled devices (CCDs), active pixel sensors (APS), and hybrid CMOS arrays. The presentation will describe the Carrier Plus goals and objectives, as well as provide details about the architecture and design. More information on the LWS program can be found at http://lws.gsfc.nasa.gov/. Business announcements for LWS/SET and program briefings are posted at http://lws-set.gsfc.nasa.gov

  12. Opsin clines in butterflies suggest novel roles for insect photopigments.

    PubMed

    Frentiu, Francesca D; Yuan, Furong; Savage, Wesley K; Bernard, Gary D; Mullen, Sean P; Briscoe, Adriana D

    2015-02-01

    Opsins are ancient molecules that enable animal vision by coupling to a vitamin-derived chromophore to form light-sensitive photopigments. The primary drivers of evolutionary diversification in opsins are thought to be visual tasks related to spectral sensitivity and color vision. Typically, only a few opsin amino acid sites affect photopigment spectral sensitivity. We show that opsin genes of the North American butterfly Limenitis arthemis have diversified along a latitudinal cline, consistent with natural selection due to environmental factors. We sequenced single nucleotide (SNP) polymorphisms in the coding regions of the ultraviolet (UVRh), blue (BRh), and long-wavelength (LWRh) opsin genes from ten butterfly populations along the eastern United States and found that a majority of opsin SNPs showed significant clinal variation. Outlier detection and analysis of molecular variance indicated that many SNPs are under balancing selection and show significant population structure. This contrasts with what we found by analysing SNPs in the wingless and EF-1 alpha loci, and from neutral amplified fragment length polymorphisms, which show no evidence of significant locus-specific or genome-wide structure among populations. Using a combination of functional genetic and physiological approaches, including expression in cell culture, transgenic Drosophila, UV-visible spectroscopy, and optophysiology, we show that key BRh opsin SNPs that vary clinally have almost no effect on spectral sensitivity. Our results suggest that opsin diversification in this butterfly is more consistent with natural selection unrelated to spectral tuning. Some of the clinally varying SNPs may instead play a role in regulating opsin gene expression levels or the thermostability of the opsin protein. Lastly, we discuss the possibility that insect opsins might have important, yet-to-be elucidated, adaptive functions in mediating animal responses to abiotic factors, such as temperature or photoperiod. PMID:25371434

  13. Living with a Star (LWS) Space Environment Testbeds (SET), Mission Carrier Overview and Capabilities

    NASA Technical Reports Server (NTRS)

    Patschke, Robert; Barth, Janet; Label, Ken; Mariano, Carolyn; Pham, Karen; Brewer, Dana; Cuviello, Michael; Kobe, David; Wu, Carl; Jarosz, Donald

    2004-01-01

    NASA has initiated the Living With a Star (LWS) Program to develop the scientific understanding to address the aspects of the Connected Sun-Earth system that affect life and society. A goal of the program is to bridge the gap between science, engineering, and user application communities. This will enable future science, operational, and commercial objectives in space and atmospheric environments by improving engineering approaches to the accommodation and/or mitigation of the effects of solar variability on technological systems. The three program elements of the LWS Program are Science Missions; Targeted Research and Technology; and Space Environment Testbeds (SETS). SET is an ideal platform for small experiments performing research on space environment effects on technologies and on the mitigation of space weather effects. A short description of the LWS Program will be given, and the SET will be described in detail, giving the mission objectives, available carrier services, and upcoming flight opportunities.

  14. Implications of the ISO LWS spectrum of the prototypical ultraluminous galaxy: ARP 220

    NASA Technical Reports Server (NTRS)

    Fischer, J.; Satyapal, S.; Luhman, M. L.; Melnick, G.; Cox, P.; Cernicharo, J.; Stacey, G. J.; Smith, H. A.; Lord, S. D.; Greenhouse, M. A.

    1997-01-01

    The low resolution far infrared spectrum of the galaxy Arp 220, obtained with the low wavelength spectrometer (LWS) onboard the Infrared Space Observatory (ISO), is presented. The spectrum is dominated by the OH, H2O, CH, NH3 and O I absorption lines. The upper limits on the far infrared fine structure lines indicate a softer radiation in Arp 220 than in starburst galaxies.

  15. Extraordinary diversity of visual opsin genes in dragonflies.

    PubMed

    Futahashi, Ryo; Kawahara-Miki, Ryouka; Kinoshita, Michiyo; Yoshitake, Kazutoshi; Yajima, Shunsuke; Arikawa, Kentaro; Fukatsu, Takema

    2015-03-17

    Dragonflies are colorful and large-eyed animals strongly dependent on color vision. Here we report an extraordinary large number of opsin genes in dragonflies and their characteristic spatiotemporal expression patterns. Exhaustive transcriptomic and genomic surveys of three dragonflies of the family Libellulidae consistently identified 20 opsin genes, consisting of 4 nonvisual opsin genes and 16 visual opsin genes of 1 UV, 5 short-wavelength (SW), and 10 long-wavelength (LW) type. Comprehensive transcriptomic survey of the other dragonflies representing an additional 10 families also identified as many as 15-33 opsin genes. Molecular phylogenetic analysis revealed dynamic multiplications and losses of the opsin genes in the course of evolution. In contrast to many SW and LW genes expressed in adults, only one SW gene and several LW genes were expressed in larvae, reflecting less visual dependence and LW-skewed light conditions for their lifestyle under water. In this context, notably, the sand-burrowing or pit-dwelling species tended to lack SW gene expression in larvae. In adult visual organs: (i) many SW genes and a few LW genes were expressed in the dorsal region of compound eyes, presumably for processing SW-skewed light from the sky; (ii) a few SW genes and many LW genes were expressed in the ventral region of compound eyes, probably for perceiving terrestrial objects; and (iii) expression of a specific LW gene was associated with ocelli. Our findings suggest that the stage- and region-specific expressions of the diverse opsin genes underlie the behavior, ecology, and adaptation of dragonflies. PMID:25713365

  16. Extraordinary diversity of visual opsin genes in dragonflies

    PubMed Central

    Futahashi, Ryo; Kawahara-Miki, Ryouka; Kinoshita, Michiyo; Yoshitake, Kazutoshi; Yajima, Shunsuke; Arikawa, Kentaro; Fukatsu, Takema

    2015-01-01

    Dragonflies are colorful and large-eyed animals strongly dependent on color vision. Here we report an extraordinary large number of opsin genes in dragonflies and their characteristic spatiotemporal expression patterns. Exhaustive transcriptomic and genomic surveys of three dragonflies of the family Libellulidae consistently identified 20 opsin genes, consisting of 4 nonvisual opsin genes and 16 visual opsin genes of 1 UV, 5 short-wavelength (SW), and 10 long-wavelength (LW) type. Comprehensive transcriptomic survey of the other dragonflies representing an additional 10 families also identified as many as 15–33 opsin genes. Molecular phylogenetic analysis revealed dynamic multiplications and losses of the opsin genes in the course of evolution. In contrast to many SW and LW genes expressed in adults, only one SW gene and several LW genes were expressed in larvae, reflecting less visual dependence and LW-skewed light conditions for their lifestyle under water. In this context, notably, the sand-burrowing or pit-dwelling species tended to lack SW gene expression in larvae. In adult visual organs: (i) many SW genes and a few LW genes were expressed in the dorsal region of compound eyes, presumably for processing SW-skewed light from the sky; (ii) a few SW genes and many LW genes were expressed in the ventral region of compound eyes, probably for perceiving terrestrial objects; and (iii) expression of a specific LW gene was associated with ocelli. Our findings suggest that the stage- and region-specific expressions of the diverse opsin genes underlie the behavior, ecology, and adaptation of dragonflies. PMID:25713365

  17. Evolutionary Dynamics of Rhodopsin Type 2 Opsins in Vertebrates

    PubMed Central

    Yokoyama, Shozo; Tada, Takashi

    2010-01-01

    Among the five groups of visual pigments in vertebrates, the rhodopsin type 2 (RH2) group shows the largest number of gene duplication events. We have isolated three intact and one nonfunctional RH2 opsin genes each from Northern lampfish (Stenobrachius leucopsarus) and scabbardfish (Lepidopus fitchi). Using the deduced amino acid sequences of these and other representative RH2 opsin genes in vertebrates, we have estimated the divergence times and evolutionary rates of amino acid substitution at various stages of RH2 opsin evolution. The results show that the duplications of the lampfish and scabbardfish RH2 opsins have occurred ∼60 and ∼30 million years ago (Ma), respectively. The evolutionary rates of RH2 opsins in the early vertebrate ancestors were ∼0.25 × 10−9/site/year, which increased to ∼1 × 10−9 to 3 × 10−9/site/year in euteleost lineages and to ∼0.3 × 10−9 to 0.5 × 10−9/site/year in coelacanth and tetrapods. PMID:19759234

  18. Plasticity of opsin gene expression in cichlids from Lake Malawi.

    PubMed

    Hofmann, Christopher M; O'Quin, Kelly E; Smith, Adam R; Carleton, Karen L

    2010-05-01

    Sensory systems play crucial roles in survival and reproduction. Therefore, sensory plasticity has important evolutionary implications. In this study, we examined retinal plasticity in five species of cichlid fish from Lake Malawi. We compared the cone opsin expression profiles of wild-caught fish to lab-reared F(1) that had been raised in a UV minus, reduced intensity light environment. All of the opsin genes that were expressed in wild-caught fish were also expressed in lab-reared individuals. However, we found statistically significant differences in relative opsin expression among all five species. The most consistent difference was in the SWS2B (violet) opsin, which was always expressed at higher levels in lab-reared individuals. Estimates of visual pigment quantum catch suggest that this change in expression would increase retinal sensitivity in the light environment of the lab. We also found that the magnitude of plasticity varied across species. These findings have important implications for understanding the genetic regulation of opsin expression and raise many interesting questions about how the cichlid visual system develops. They also suggest that sensory plasticity may have facilitated the ecological diversification of cichlids in Lake Malawi. PMID:20374487

  19. Non-visual photoreception by a variety of vertebrate opsins.

    PubMed

    Kojima, D; Fukada, Y

    1999-01-01

    Extraretinal photoreceptors in animals are involved in a variety of physiological functions such as photo-entrainment of circadian rhythm, photoperiodicity and body colour change. We have identified pinopsin in the chicken pineal gland as a typical 'non-visual' photoreceptive molecule. Pinopsin with bound 11-cis-retinal shows a blue-light sensitivity (lambda max = 468 nm), and it may play a role in synchronizing the phase of the endogenous circadian oscillator with an environmental dark-light cycle. Pinopsin is not a unique pineal opsin in animals. In the zebrafish, we have detected expression of two rhodopsin genes, the nucleotide sequences of which are very similar but distinct from each other. One is canonical rhodopsin expressed in the retina, and the other is expressed in the pineal gland. The latter gene is widely distributed among teleosts, and we named it 'exo-rhodopsin' after extraocular rhodopsin. On the other hand, our effort to identify the 'deep brain opsin' responsible for the photoperiodic gonadal response resulted in the identification of two kinds of opsins; pinopsin in the toad anterior preoptic nucleus and rhodopsin in the pigeon lateral septum. Both of these opsins are localized in the cerebrospinal fluid-contacting neurons in the brain of the two animals. We also identified VAL opsin in zebrafish retinal horizontal cells, which have not been considered as photoreceptive cells. It has become evident that animals employ a wide variety of photoreceptive molecules for 'non-visual' purposes. PMID:10614056

  20. The Evolution and Expression of the Moth Visual Opsin Family

    PubMed Central

    Fu, Xiaowei; Murphy, Robert W.; Wu, Kongming

    2013-01-01

    Because visual genes likely evolved in response to their ambient photic environment, the dichotomy between closely related nocturnal moths and diurnal butterflies forms an ideal basis for investigating their evolution. To investigate whether the visual genes of moths are associated with nocturnal dim-light environments or not, we cloned long-wavelength (R), blue (B) and ultraviolet (UV) opsin genes from 12 species of wild-captured moths and examined their evolutionary functions. Strong purifying selection appeared to constrain the functions of the genes. Dark-treatment altered the levels of mRNA expression in Helicoverpa armigera such that R and UV opsins were up-regulated after dark-treatment, the latter faster than the former. In contrast, B opsins were not significantly up-regulated. Diel changes of opsin mRNA levels in both wild-captured and lab-reared individuals showed no significant fluctuation within the same group. However, the former group had significantly elevated levels of expression compared with the latter. Consequently, environmental conditions appeared to affect the patterns of expression. These findings and the proportional expression of opsins suggested that moths potentially possessed color vision and the visual system played a more important role in the ecology of moths than previously appreciated. This aspect did not differ much from that of diurnal butterflies. PMID:24205129

  1. Neuronal Organization of Deep Brain Opsin Photoreceptors in Adult Teleosts

    PubMed Central

    Hang, Chong Yee; Kitahashi, Takashi; Parhar, Ishwar S.

    2016-01-01

    Biological impacts of light beyond vision, i.e., non-visual functions of light, signify the need to better understand light detection (or photoreception) systems in vertebrates. Photopigments, which comprise light-absorbing chromophores bound to a variety of G-protein coupled receptor opsins, are responsible for visual and non-visual photoreception. Non-visual opsin photopigments in the retina of mammals and extra-retinal tissues of non-mammals play an important role in non-image-forming functions of light, e.g., biological rhythms and seasonal reproduction. This review highlights the role of opsin photoreceptors in the deep brain, which could involve conserved neurochemical systems that control different time- and light-dependent physiologies in in non-mammalian vertebrates including teleost fish. PMID:27199680

  2. Vertebrate ancient opsin and melanopsin: divergent irradiance detectors.

    PubMed

    Davies, Wayne L; Hankins, Mark W; Foster, Russell G

    2010-11-01

    Both vertebrates and invertebrates respond to light by utilising a wide-ranging array of photosensory systems, with diverse photoreceptor organs expressing a characteristic photopigment, itself consisting of an opsin apoprotein linked to a light-sensitive retinoid chromophore based on vitamin A. In the eye, the pigments expressed in both cone and rod photoreceptors have been studied in great depth and mediate contrast perception, measurement of the spectral composition of environmental light, and thus classical image forming vision. By contrast, the molecular basis for non-visual and extraocular photoreception is far less understood; however, two photopigment genes have become the focus of much study, the vertebrate ancient (va) opsin and melanopsin (opn4). In this review, we discuss the history of discovery for each gene, as well as focusing on the evolution, expression profile, functional role and broader physiological significance of each photopigment. Recently, it has been suggested independently by Arendt et al. and Lamb that an ancestral opsin bifurcated in early metazoans and evolved into two quite different photopigments, one expressed in rhabdomeric photoreceptors and the other in ciliary photoreceptors. This interpretation of the evolution of the metazoan eye has provided a powerful framework for understanding photobiological organization. Their proposal, however, does not encompass all current experimental observations that would be consistent with what we term a central "Evolution of Photosensory Opsins with Common Heredity (EPOCH)" hypothesis to explain the complexity of animal photosensory systems. Clearly, many opsin genes (e.g. va opsin) simply do not fit neatly within this scheme. Thus, the review concludes with a discussion of these anomalies and their context regarding the phylogeny of photoreceptor and photopigment development. PMID:20922256

  3. Parallel evolution of opsin gene expression in African cichlid fishes.

    PubMed

    O'Quin, Kelly E; Hofmann, Christopher M; Hofmann, Hans A; Carleton, Karen L

    2010-12-01

    Phenotypic evolution may occur either through alterations to the structure of protein-coding genes or their expression. Evidence for which of these two mechanisms more commonly contribute to the evolution of a phenotype can be garnered from examples of parallel and convergent evolution. The visual system of East African cichlid fishes is an excellent system with which to address this question. Cichlid fishes from Lakes Malawi (LM) and Victoria together exhibit three diverse palettes of coexpressed opsins and several important protein-coding mutations that both shift spectral sensitivity. Here we assess both opsin expression and protein-coding diversity among cichlids from a third rift lake, Lake Tanganyika (LT). We found that Tanganyikan cichlids exhibit three palettes of coexpressed opsins that largely overlap the short-, middle-, and long-wavelength-sensitive palettes of LM cichlids. Bayesian phenotypic clustering and ancestral state reconstructions both support the parallel evolution of the short- and middle-wavelength palettes among cichlids from LT and LM. In each case, these transitions occurred from different ancestors that expressed the same long-wavelength palette. We also identified similar but distinct patterns of correlated evolution between opsin expression, diet, and lens transmittance among cichlids from LT and LM as well. In contrast to regulatory changes, we identified few functional or potentially functional mutations in the protein-coding sequences of three variable opsins, with the possible exception of the SWS1 (ultraviolet) opsin. These results underscore the important contribution that gene regulation can make to rapid phenotypic evolution and adaptation. PMID:20601410

  4. Opsin phylogeny and evolution: a model for blue shifts in wavelength regulation.

    PubMed

    Chang, B S; Crandall, K A; Carulli, J P; Hartl, D L

    1995-03-01

    The vast diversity in spectral sensitivities in the vision of many organisms is mediated mostly (although not entirely) through variation in the photosensitive visual pigments (opsins) of the eye. Specifically, shifts in absorption maxima of visual pigments are thought to be a result of interactions within the binding pocket of the opsin, between amino acid side chains and the retinal chromophore, However, it has proven difficult to identify specific amino acid residues important in determining wavelength absorption maxima, especially for some of the short wavelength (blue) opsins. In this paper, a comparative phylogenetic approach was applied to opsin protein sequence data to identify residues important in opsin wavelength regulation. In essence, this approach consisted of interpreting evolutionary history as a series of experiments in which natural selection has repeatedly favored amino acid replacements of certain residues to shift the opsin absorption spectra to either shorter or longer wavelengths. Opsin protein sequences were obtained from GenBank, aligned, and used to reconstruct a phylogenetic tree. Amino acid replacements were traced along the branches of this opsin tree, focusing only on residues likely to reside within the chromophore-binding pocket. A number of functionally convergent, nonconservative amino acid replacements in independently evolved opsins with similar shifts in spectral properties were identified. In short, reconstruction of the phylogeny of the opsin molecule allowed us to track amino acid substitutions in specific sites within the opsin and to target those particular substitutions that are repeatedly associated with marked changes in peak absorbance, shifting the spectral sensitivity of the opsin toward shorter or longer wavelengths. Based on these results, we propose a model for blue shifts of opsin absorption spectra. Amino acid replacements of four polar and charged residues near the protonated Schiff base (SBH+) end of the chromophore are proposed to result in blue shifts of the opsin absorption spectra. This model may explain some of the diversity of blue opsins apparent in both vertebrates and invertebrates. PMID:7620634

  5. Variations in Opsin Coding Sequences Cause X-Linked Cone Dysfunction Syndrome with Myopia and Dichromacy

    PubMed Central

    McClements, Michelle; Davies, Wayne I. L.; Michaelides, Michel; Young, Terri; Neitz, Maureen; MacLaren, Robert E.; Moore, Anthony T.; Hunt, David M.

    2013-01-01

    Purpose. To determine the role of variant L opsin haplotypes in seven families with Bornholm Eye Disease (BED), a cone dysfunction syndrome with dichromacy and myopia. Methods. Analysis of the opsin genes within the L/M opsin array at Xq28 included cloning and sequencing of an exon 3-5 gene fragment, long range PCR to establish gene order, and quantitative PCR to establish gene copy number. In vitro expression of normal and variant opsins was performed to examine cellular trafficking and spectral sensitivity of pigments. Results. All except one of the BED families possessed L opsin genes that contained a rare exon 3 haplotype. The exception was a family with the deleterious Cys203Arg substitution. Two rare exon 3 haplotypes were found and, where determined, these variant opsin genes were in the first position in the array. In vitro expression in transfected cultured neuronal cells showed that the variant opsins formed functional pigments, which trafficked to the cell membranes. The variant opsins were, however, less stable than wild type. Conclusions. It is concluded that the variant L opsin haplotypes underlie BED. The reduction in the amount of variant opsin produced in vitro compared with wild type indicates a possible disease mechanism. Alternatively, the recently identified defective splicing of exon 3 of the variant opsin transcript may be involved. Both mechanisms explain the presence of dichromacy and cone dystrophy. Abnormal pigment may also underlie the myopia that is invariably present in BED subjects. PMID:23322568

  6. Restoration of Vision with Ectopic Expression of Human Rod Opsin

    PubMed Central

    Cehajic-Kapetanovic, Jasmina; Eleftheriou, Cyril; Allen, Annette E.; Milosavljevic, Nina; Pienaar, Abigail; Bedford, Robert; Davis, Katherine E.; Bishop, Paul N.; Lucas, Robert J.

    2015-01-01

    Summary Many retinal dystrophies result in photoreceptor loss, but the inner retinal neurons can survive, making them potentially amenable to emerging optogenetic therapies. Here, we show that ectopically expressed human rod opsin, driven by either a non-selective or ON-bipolar cell-specific promoter, can function outside native photoreceptors and restore visual function in a mouse model of advanced retinal degeneration. Electrophysiological recordings from retinal explants and the visual thalamus revealed changes in firing (increases and decreases) induced by simple light pulses, luminance increases, and naturalistic movies in treated mice. These responses could be elicited at light intensities within the physiological range and substantially below those required by other optogenetic strategies. Mice with rod opsin expression driven by the ON-bipolar specific promoter displayed behavioral responses to increases in luminance, flicker, coarse spatial patterns, and elements of a natural movie at levels of contrast and illuminance (≈50–100 lux) typical of natural indoor environments. These data reveal that virally mediated ectopic expression of human rod opsin can restore vision under natural viewing conditions and at moderate light intensities. Given the inherent advantages in employing a human protein, the simplicity of this intervention, and the quality of vision restored, we suggest that rod opsin merits consideration as an optogenetic actuator for treating patients with advanced retinal degeneration. PMID:26234216

  7. Expression Dynamics and Protein Localization of Rhabdomeric Opsins in Platynereis Larvae

    PubMed Central

    Randel, Nadine; Bezares-Calderón, Luis A.; Gühmann, Martin; Shahidi, Réza; Jékely, Gáspár

    2013-01-01

    The larval stages of polychaete annelids are often responsive to light and can possess one to six eyes. The early trochophore larvae of the errant annelid Platynereis dumerilii have a single pair of ventral eyespots, whereas older nectochaete larvae have an additional two pairs of dorsal eyes that will develop into the adult eyes. Early Platynereis trochophores show robust positive phototaxis starting on the first day of development. Even though the mechanism of phototaxis in Platynereis early trochophore larvae is well understood, no photopigment (opsin) expression has yet been described in this stage. In late trochophore larvae, a rhabdomeric-type opsin, r-opsin1, expressed in both the eyespots and the adult eyes has already been reported. Here, we identify another Platynereis rhabdomeric opsin, r-opsin3, that is expressed in a single photoreceptor in the eyespots in early trochophores, suggesting that it mediates early larval phototaxis. We also show that r-opsin1 and r-opsin3 are expressed in adjacent photoreceptor cells in the eyespots in later stages, indicating that a second eyespot-photoreceptor differentiates in late trochophore larvae. Using serial transmission electron microscopy (TEM), we identified and reconstructed both photoreceptors and a pigment cell in the late larval eyespot. We also characterized opsin expression in the adult eyes and found that the two opsins co-express there in several photoreceptor cells. Using antibodies recognizing r-opsin1 and r-opsin3 proteins, we demonstrate that both opsins localize to the rhabdomere in all six eyes. In addition, we found that r-opsin1 mRNA is localized to, and translated in, the projections of the adult eyes. The specific changes we describe in opsin transcription and translation and in the cellular complement suggest that the six larval eyes undergo spectral and functional maturation during the early planktonic phase of the Platynereis life cycle. PMID:23667045

  8. Characterisation and localisation of the opsin protein repertoire in the brain and retinas of a spider and an onychophoran

    PubMed Central

    2013-01-01

    Background Opsins have been found in the majority of animals and their most apparent functions are related to vision and light-guided behaviour. As an increasing number of sequences have become available it has become clear that many opsin-like transcripts are expressed in tissues other than the eyes. Opsins can be divided into three main groups: rhabdomeric opsins (r-opsins), ciliary opsins (c-opsins) and group 4 opsins. In arthropods, the main focus has been on the r-opsins involved in vision. However, with increased sequencing it is becoming clear that arthropods also possess opsins of the c-type, group 4 opsins and the newly discovered arthropsins but the functions of these opsins are unknown in arthropods and data on their localisation is limited or absent. Results We identified opsins from the spider Cupiennius salei and the onychophoran Euperipatoides kanangrensis and characterised the phylogeny and localisation of these transcripts. We recovered all known visual opsins in C. salei, and in addition found a peropsin, a c-opsin and an opsin resembling Daphnia pulex arthropsin. The peropsin was expressed in all eye types except the anterior median eyes. The arthropsin and the c-opsin were expressed in the central nervous system but not the eyes. In E. kanangrensis we found: a c-opsin; an opsin resembling D. pulex arthropsins; and an r-opsin with high sequence similarity to previously published onychophoran onychopsins. The E. kanangrensis c-opsin and onychopsin were expressed in both the eyes and the brain but the arthropsin only in the brain. Conclusion Our novel finding that opsins of both the ciliary and rhabdomeric type are present in the onychophoran and a spider suggests that these two types of opsins were present in the last common ancestor of the Onychophora and Euarthropoda. The expression of the c-opsin in the eye of an onychophoran indicates that c-opsins may originally have been involved in vision in the arthropod clade. The lack of c-opsin expression in the spider retina suggests that the role for c-opsin in vision was lost in the euarthropods. Our discovery of arthropsin in onychophorans and spiders dates the emergence of arthropsin to the common ancestor of Onychophora and Euarthropoda and their expression in the brain suggests a non-visual function. PMID:24010579

  9. Chemical name to structure: OPSIN, an open source solution.

    PubMed

    Lowe, Daniel M; Corbett, Peter T; Murray-Rust, Peter; Glen, Robert C

    2011-03-28

    We have produced an open source, freely available, algorithm (Open Parser for Systematic IUPAC Nomenclature, OPSIN) that interprets the majority of organic chemical nomenclature in a fast and precise manner. This has been achieved using an approach based on a regular grammar. This grammar is used to guide tokenization, a potentially difficult problem in chemical names. From the parsed chemical name, an XML parse tree is constructed that is operated on in a stepwise manner until the structure has been reconstructed from the name. Results from OPSIN on various computer generated name/structure pair sets are presented. These show exceptionally high precision (99.8%+) and, when using general organic chemical nomenclature, high recall (98.7-99.2%). This software can serve as the basis for future open source developments of chemical name interpretation. PMID:21384929

  10. LWS TR&T Project on the CME - ICME Connection: A Progress Report

    NASA Astrophysics Data System (ADS)

    Georgoulis, M. K.

    2006-12-01

    During the second year of the above LWS TR&T Grant, we report on our efforts and progress. The Project highlights and will continue to uncover the wealth of eruptive diagnostics that can be obtained by studying vector (but also line-of-sight) magnetograms of solar active regions. Here we focus on one particular aspect of active-region physics: the elusive relation between the free magnetic energy and the magnetic helicity budget of eruptive and non-eruptive active regions. Besides shedding light into the eruption process itself, this analysis can uncover the true relation between the magnetic structures of the propagating ICMEs and that of the progenitor CMEs and the source active regions. This, as the title suggests, is the ultimate objective of the Project.

  11. Modeling Active Region Evolution - A New LWS TR and T Strategic Capability Model Suite

    NASA Technical Reports Server (NTRS)

    MacNeice, Peter

    2012-01-01

    In 2006 the LWS TR&T Program funded us to develop a strategic capability model of slowly evolving coronal active regions. In this poster we report on the overall design, and status of our new modeling suite. Our design features two coronal field models, a non-linear force free field model and a global 3D MHD code. The suite includes supporting tools and a user friendly GUI which will enable users to query the web for relevant magnetograms, download them, process them to synthesize a sequence of photospheric magnetograms and associated photospheric flow field which can then be applied to drive the coronal model innner boundary, run the coronal models and finally visualize the results.

  12. 2006 LWS TR&T Solar Wind Focused Science Topic Team: Overview of Current Results

    NASA Astrophysics Data System (ADS)

    Miralles, M. P.; LWS Tr&T Solar Wind Fst Team

    2008-12-01

    We present a summary of the research conducted by the members of the 2006 LWS TR&T Solar Wind Focused Science Topic (FST) Team on the physical processes that heat and accelerate the solar wind. The Team applied a combination of theoretical studies, numerical simulations, and observations for their investigation of the role of energy sources and kinetic mechanisms responsible for the heating and acceleration of the solar wind. In particular, the FST Team examined magnetic reconnection, waves, and turbulence as possible heating mechanisms. Plasma properties and their evolution over the solar cycle, determined from the analysis of remote and in situ measurements of solar wind source regions and streams, are being used to constrain the models. The consistency of candidate theoretical models with existing observational data for the solar wind will be discussed.

  13. The ISO-LWS map of the Serpens cloud core. II. The line spectra

    NASA Astrophysics Data System (ADS)

    Larsson, B.; Liseau, R.; Men'shchikov, A. B.

    2002-05-01

    We present spectrophotometric ISO imaging with the LWS and the CAM-CVF of the Serpens molecular cloud core. The LWS map is centred on the far infrared and submillimetre source FIRS 1/SMM 1 and its size is 8',x 8'. The fine structure line emission in [O I] 63 mu m and [C II] 157 mu m is extended on the arcminute scale and can be successfully modelled to originate in a PDR with G0 = 15 +/- 10 and n(H2) in the range of (104-105) cm-3. Extended emission might also be observed in the rotational line emission of H2O and high-J CO. However, lack of sufficient angular resolution prevents us from excluding the possibility that the emssion regions of these lines are point like, which could be linked to the embedded objects SMM 9/S 68 and SMM 4. Toward the Class 0 source SMM 1, the LWS observations reveal, in addition to fine structure line emission, a rich spectrum of molecular lines, superposed onto a strong, optically thick dust continuum (Larsson et al. \\cite{Lar00}). The sub-thermally excited and optically thick CO, H2O and OH lines are tracing an about 103 AU source with temperatures higher than 300 K and densities above 106 cm-3 (M=0.01 Msun). The molecular abundances, X=N(mol)/N(H2), are X=(1, 0.1, 0.02, ge 0.025) x 10-4 for CO, H2O, OH and 13CO, respectively. Our data are consistent with an ortho-to-para ratio of 3 for H2O. OH appears highly overabundant, which we tentatively ascribe to an enhanced (X-ray) ionisation rate in the Serpens cloud core (zeta >> 10-18 s-1). We show that geometry is of concern for the correct interpretation of the data and based on 2D-radiative transfer modelling of the disk/torus around SMM 1, which successfully reproduces the entire observed SED and the observed line profiles of low-to-mid-J CO isotopomers, we can exclude the disk to be the source of the LWS-molecular line emission. The same conclusion applies to models of dynamical collapse (``inside-out'' infall). The 6{' '} pixel resolution of the CAM-CVF permits us to see that the region of rotational H2 emission is offset from SMM 1 by 30{' '}, at position angle 340deg, which is along the known jet flow from the Class 0 object. This H2 gas is extinguished by AV = 4.5 mag and at a temperature of 10310 K, which suggests that the heating of the gas is achieved through relatively slow shocks. Although we are not able to establish any firm conclusion regarding the detailed nature of the shock waves, our observations of the molecular line emission from SMM 1 are to a limited extent explainable in terms of an admixture of J-shocks and of C-shocks, the latter with speeds of about (15-20) km s-1, whereas dynamical infall is not directly revealed by our data. Based on observations with ISO, an ESA project with instruments funded by ESA Member States (especially the PI countries: France, Germany, The Netherlands and the UK) and with the participation of ISAS and NASA.

  14. ISO-LWS observations of Herbig Ae/Be stars. I. Fine structure lines

    NASA Astrophysics Data System (ADS)

    Lorenzetti, D.; Tommasi, E.; Giannini, T.; Nisini, B.; Benedettini, M.; Pezzuto, S.; Strafella, F.; Barlow, M.; Clegg, P. E.; Cohen, M.; di Giorgio, A. M.; Liseau, R.; Molinari, S.; Palla, F.; Saraceno, P.; Smith, H. A.; Spinoglio, L.; White, G. J.

    1999-06-01

    We present the results of the first spectrophotometric survey of a sample of eleven Herbig Ae/Be stars (HAEBE) obtained with the Long Wavelength Spectrometer (LWS) on board the Infrared Space Observatory (ISO). The [OI] 63mu m and the [CII] 158mu m lines are observed in all the investigated sources, while the [OI] 145mu m transition, due to its relative faintness, sometimes remains undetected. By comparing line intensity ratios with model predictions, photodissociation, due to the UV photons from the central star, results the dominating excitation mechanism although contributions of C-shocks to the [OI] emission cannot be ruled out. A clear example for the presence of a photodissociation region (PDR) illuminated by an HAEBE is shown by LWS spectroscopic mapping of NGC 7129. Some diagnostic probes of the radiation field and density are provided for the objects in our sample: these substantially agree with the known characteristics of both the star and its circumstellar environment, although the observed ratio [OI]63/[OI]145 tends to be smaller than predicted by PDR models. The most likely explanation for this behaviour is self-absorption at 63mu m by cold atomic oxygen. Fine structure lines of the ionised species [OIII], [NII] were detected whenever the star had a spectral type of B0 or earlier; in particular, around the star CoD-42(deg) 11721, besides a compact HII region, evidence is given for an extended low electron density ionised region. Finally, molecular line emission is associated with stars powering a CO outflow, and clumpy PDR models, better than C-shock models, predict for them relative cooling (CO vs OI and CO vs OH) similar to the observed ones. Based on observations with ISO, an ESA project with instruments funded by ESA Member States and with the participation of ISAS and NASA}

  15. Cone opsin determines the time course of cone photoreceptor degeneration in Leber congenital amaurosis.

    PubMed

    Zhang, Tao; Zhang, Ning; Baehr, Wolfgang; Fu, Yingbin

    2011-05-24

    Mutations in RPE65 or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal recycling and cause Leber congenital amaurosis (LCA), the most severe retinal dystrophy in early childhood. We used Lrat(-)(/-), a murine model for LCA, to investigate the mechanism of rapid cone degeneration. Although both M and S cone opsins mistrafficked as reported previously, mislocalized M-opsin was degraded whereas mislocalized S-opsin accumulated in Lrat(-)(/-) cones before the onset of massive ventral/central cone degeneration. As the ventral and central retina express higher levels of S-opsin than the dorsal retina in mice, our results may explain why ventral and central cones degenerate more rapidly than dorsal cones in Rpe65(-)(/-) and Lrat(-)(/-) LCA models. In addition, human blue opsin and mouse S-opsin, but not mouse M-opsin or human red/green opsins, aggregated to form cytoplasmic inclusions in transfected cells, which may explain why blue cone function is lost earlier than red/green-cone function in patients with LCA. The aggregation of short-wavelength opsins likely caused rapid cone degenerations through an endoplasmic reticulum stress pathway, as demonstrated in both the Lrat(-)(/-) retina and transfected cells. Replacing rhodopsin with S-opsin in Lrat(-)(/-) rods resulted in mislocalization and aggregation of S-opsin in the inner segment and the synaptic region of rods, ER stress, and dramatically accelerated rod degeneration. Our results demonstrate that cone opsins play a major role in determining the degeneration rate of photoreceptors in LCA. PMID:21555576

  16. Cone opsin determines the time course of cone photoreceptor degeneration in Leber congenital amaurosis

    PubMed Central

    Zhang, Tao; Zhang, Ning; Baehr, Wolfgang; Fu, Yingbin

    2011-01-01

    Mutations in RPE65 or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal recycling and cause Leber congenital amaurosis (LCA), the most severe retinal dystrophy in early childhood. We used Lrat−/−, a murine model for LCA, to investigate the mechanism of rapid cone degeneration. Although both M and S cone opsins mistrafficked as reported previously, mislocalized M-opsin was degraded whereas mislocalized S-opsin accumulated in Lrat−/− cones before the onset of massive ventral/central cone degeneration. As the ventral and central retina express higher levels of S-opsin than the dorsal retina in mice, our results may explain why ventral and central cones degenerate more rapidly than dorsal cones in Rpe65−/− and Lrat−/− LCA models. In addition, human blue opsin and mouse S-opsin, but not mouse M-opsin or human red/green opsins, aggregated to form cytoplasmic inclusions in transfected cells, which may explain why blue cone function is lost earlier than red/green-cone function in patients with LCA. The aggregation of short-wavelength opsins likely caused rapid cone degenerations through an endoplasmic reticulum stress pathway, as demonstrated in both the Lrat−/− retina and transfected cells. Replacing rhodopsin with S-opsin in Lrat−/− rods resulted in mislocalization and aggregation of S-opsin in the inner segment and the synaptic region of rods, ER stress, and dramatically accelerated rod degeneration. Our results demonstrate that cone opsins play a major role in determining the degeneration rate of photoreceptors in LCA. PMID:21555576

  17. Selective Protonation of Acidic Residues Triggers Opsin Activation.

    PubMed

    Lans, Isaias; Dalton, James A R; Giraldo, Jesús

    2015-07-30

    Rhodopsin, the visual pigment in the retina, is a Class A G protein-coupled receptor (GPCR) covalently bound to retinal chromophore. In dark conditions, retinal is in the cis-isomeric state, stabilizing the rhodopsin inactive state as an inverse agonist. After light absorption, retinal undergoes an isomerization photoreaction to trans-retinal, which includes a conformational change of the receptor to its active state. In the absence of retinal, the apoprotein opsin presents a low level of constitutive activity, which depends on pH, with higher propensity of activation at acidic pH. To examine the effect and the underlying mechanism that protonation may have on opsin activation, a number of MD simulations were run varying the number and identity of acidic residues selected for protonation. Results show that the combined protonation of D83, E113, and E247 is of special relevance for the induction of receptor activation. Subsequent conformational analysis of the MD trajectories provides a structural mechanistic insight into the opsin activation process. Furthermore, because protonation seems to be a determining step in the activation of other GPCRs, the methodology and rationale used herein can be extended to mechanistic studies of GPCRs in general. PMID:26140747

  18. True blue: S-opsin is widely expressed in different animal species.

    PubMed

    Amann, B; Hirmer, S; Hauck, S M; Kremmer, E; Ueffing, M; Deeg, C A

    2014-02-01

    Colour vision in animals is an interesting, fascinating subject. In this study, we examined a wide variety of species for expression of S-opsin (blue sensitive) and M-/L-opsin (green-red sensitive) in retinal cones using two novel monoclonal antibodies specific for peptides from human opsins. Mouse, rat and hare did not express one of the investigated epitopes, but we could clearly prove existence of cones through peanut agglutinin labelling. Retinas of guinea pig, dog, wolf, marten, cat, roe deer, pig and horse were positive for S-opsin, but not for M-/L-opsin. Nevertheless all these species are clearly at least dichromats, because we could detect further S-opsin negative cones by labelling with cone arrestin specific antibody. In contrast, pheasant and char had M-/L-opsin positive cones, but no S-opsin expressing cones. Sheep, cattle, monkey, men, pigeon, duck and chicken were positive for both opsins. Visual acuity analyzed through density of retinal ganglion cells revealed least visual discrimination by horses and highest resolution in pheasant and pigeon. Most mammals studied are dichromats with visual perception similar to red-green blind people. PMID:23173557

  19. Co-expression of three opsins in cone photoreceptors of the salamander, Ambystoma tigrinum

    PubMed Central

    Isayama, Tomoki; Chen, Ying; Kono, Masahiro; Fabre, Eduard; Slavsky, Michael; DeGrip, Willem J.; Ma, Jian-Xing; Crouch, Rosalie K.; Makino, Clint L.

    2014-01-01

    Whereas more than one type of visual opsin is present in the retina of most vertebrates, it was thought that each type of photoreceptor expressed only one opsin. However, evidence has accumulated that some photoreceptors contain more than one opsin, in many cases as a result of a developmental transition from the expression of one opsin to another. The salamander UV-sensitive (UV) cone is particularly notable because it contains three opsins (Makino and Dodd, 1996; J Gen Physiol 108:27–34). Two opsin types are expressed at levels more than a hundred times lower than that of the primary opsin. Here, immunohistochemical experiments identified the primary component as a UV cone opsin and the two minor components as the short wavelength-sensitive (S) and long wavelength-sensitive (L) cone opsins. Based on single cell recordings of 156 photoreceptors, the presence of three components in UV cones of hatchlings and terrestrial adults ruled out a developmental transition. There was no evidence for multiple opsin types within rods or S cones. But immunohistochemistry and partial bleaching in conjunction with single cell recording revealed that both single and double L cones contained low levels of short wavelength-sensitive pigments in addition to the main L visual pigment. These results raise the possibility that co-expression of multiple opsins in other vertebrates was overlooked because a minor component absorbing at short wavelengths was masked by the main visual pigment or because the expression level of a component absorbing at long wavelengths was exceedingly low. PMID:24374736

  20. Coexpression of three opsins in cone photoreceptors of the salamander Ambystoma tigrinum.

    PubMed

    Isayama, Tomoki; Chen, Ying; Kono, Masahiro; Fabre, Eduard; Slavsky, Michael; DeGrip, Willem J; Ma, Jian-Xing; Crouch, Rosalie K; Makino, Clint L

    2014-07-01

    Although more than one type of visual opsin is present in the retina of most vertebrates, it was thought that each type of photoreceptor expresses only one opsin. However, evidence has accumulated that some photoreceptors contain more than one opsin, in many cases as a result of a developmental transition from the expression of one opsin to another. The salamander UV-sensitive (UV) cone is particularly notable because it contains three opsins (Makino and Dodd [1996] J Gen Physiol 108:27-34). Two opsin types are expressed at levels more than 100 times lower than the level of the primary opsin. Here, immunohistochemical experiments identified the primary component as a UV cone opsin and the two minor components as the short wavelength-sensitive (S) and long wavelength-sensitive (L) cone opsins. Based on single-cell recordings of 156 photoreceptors, the presence of three components in UV cones of hatchlings and terrestrial adults ruled out a developmental transition. There was no evidence for multiple opsin types within rods or S cones, but immunohistochemistry and partial bleaching in conjunction with single-cell recording revealed that both single and double L cones contained low levels of short wavelength-sensitive pigments in addition to the main L visual pigment. These results raise the possibility that coexpression of multiple opsins in other vertebrates was overlooked because a minor component absorbing at short wavelengths was masked by the main visual pigment or because the expression level of a component absorbing at long wavelengths was exceedingly low. PMID:24374736

  1. Opsin sequences of the rod visual pigments in two species of Poeciliid fish

    PubMed

    Archer; Hirano

    1997-07-01

    The rod opsin sequences from Gambusia affinis holbrooki and Poecilia reticulata were cloned and sequenced. The opsin sequences were found to be 96·8% identical, reflecting the similarity of the rod visual pigment absorbances in these two Poeciliid fish. PMID:9236103

  2. Opsin evolution and expression in Arthropod compound Eyes and Ocelli: Insights from the cricket Gryllus bimaculatus

    PubMed Central

    2012-01-01

    Background Opsins are key proteins in animal photoreception. Together with a light-sensitive group, the chromophore, they form visual pigments which initiate the visual transduction cascade when photoactivated. The spectral absorption properties of visual pigments are mainly determined by their opsins, and thus opsins are crucial for understanding the adaptations of animal eyes. Studies on the phylogeny and expression pattern of opsins have received considerable attention, but our knowledge about insect visual opsins is still limited. Up to now, researchers have focused on holometabolous insects, while general conclusions require sampling from a broader range of taxa. We have therefore investigated visual opsins in the ocelli and compound eyes of the two-spotted cricket Gryllus bimaculatus, a hemimetabolous insect. Results Phylogenetic analyses place all identified cricket sequences within the three main visual opsin clades of insects. We assign three of these opsins to visual pigments found in the compound eyes with peak absorbances in the green (515 nm), blue (445 nm) and UV (332 nm) spectral range. Their expression pattern divides the retina into distinct regions: (1) the polarization-sensitive dorsal rim area with blue- and UV-opsin, (2) a newly-discovered ventral band of ommatidia with blue- and green-opsin and (3) the remainder of the compound eye with UV- and green-opsin. In addition, we provide evidence for two ocellar photopigments with peak absorbances in the green (511 nm) and UV (350 nm) spectral range, and with opsins that differ from those expressed in the compound eyes. Conclusions Our data show that cricket eyes are spectrally more specialized than has previously been assumed, suggesting that similar adaptations in other insect species might have been overlooked. The arrangement of spectral receptor types within some ommatidia of the cricket compound eyes differs from the generally accepted pattern found in holometabolous insect taxa and awaits a functional explanation. From the opsin phylogeny, we conclude that gene duplications, which permitted differential opsin expression in insect ocelli and compound eyes, occurred independently in several insect lineages and are recent compared to the origin of the eyes themselves. PMID:22935102

  3. Functional properties of opsins and their contribution to light-sensing physiology.

    PubMed

    Terakita, Akihisa; Nagata, Takashi

    2014-10-01

    Many animals have developed systems for sensing environmental conditions during evolution. In sensory cells, receptor molecules are responsible for their sensing abilities. In light sensing, most animals capture light information via rhodopsin-like photoreceptive proteins known as opsin-based pigments. A body of evidence from comparisons of amino acid sequences and in vitro experiments demonstrates that opsins have phylogenetically and functionally diversified during evolution and suggests that the phylogenetic diversity in opsins correlates with the variety of molecular properties of opsin-based pigments. In this review, we discuss the various molecular properties of opsin-based pigments and their contribution to light-sensing ability by providing two examples: i) contribution of photoregeneration ability and Chromophore retinal binding property of an Opn3 homolog to non-visual photoreception, and ii) contribution of an absorption characteristic of a visual pigment to depth perception in jumping spiders. PMID:25284384

  4. Recent Results of the 2005 LWS TR&T Focus Team for Solar Energetic Particles

    NASA Astrophysics Data System (ADS)

    Desai, M. I.; Cohen, C. M.; Smith, C. W.; Lee, M. A.; Litvinenko, Y.; Reames, D. V.; Ng, C. K.; Tylka, A. J.; Kota, J.; Giacalone, J.; Jokipii, J. R.; Sokolov, I.; Gombosi, T.; Roussev, I. I.; Li, G.; Zank, G. P.; Tessein, J.

    2006-12-01

    Shock waves driven by coronal mass ejections are presently believed to be responsible for producing large gradual solar energetic particle (SEP) events that can pose significant radiation hazard for humans and technological systems near Earth. However, our present ability to accurately predict various properties of SEP events (e.g., peak intensities, energy spectra, and composition) is somewhat limited. Reliable prediction of these properties depends on a multitude of observational and theoretical cross-disciplinary studies that include: (1) Understanding and modeling the initiation and propagation of fast CMEs and the evolution of shock characteristics as they travel through the corona and interplanetary medium, (2) Characterizing the ambient coronal and solar wind plasma, the solar and interplanetary magnetic field, and the suprathermal ion population through which the CMEs and their shocks propagate en route to Earth, and (3) Developing a detailed understanding of particle injection and acceleration at CME-driven shocks and their subsequent transport out to 1 AU. Such studies are being routinely conducted both, individually and collectively by members of the 2005 LWS TR&T Focus Team for SEPs. In this talk we will summarize the SEP Team's ongoing efforts, highlight the key new results, and discuss some of the main scientific challenges that we need to overcome in order to improve current understanding of the physical processes occurring in large SEP events.

  5. Three Different Cone Opsin Gene Array Mutational Mechanisms with GenotypePhenotype Correlation and Functional Investigation of Cone Opsin Variants

    PubMed Central

    Gardner, Jessica C; Liew, Gerald; Quan, Ying-Hua; Ermetal, Burcu; Ueyama, Hisao; Davidson, Alice E; Schwarz, Nele; Kanuga, Naheed; Chana, Ravinder; Maher, Eamonn R; Webster, Andrew R; Holder, Graham E; Robson, Anthony G; Cheetham, Michael E; Liebelt, Jan; Ruddle, Jonathan B; Moore, Anthony T; Michaelides, Michel; Hardcastle, Alison J

    2014-01-01

    Mutations in the OPN1LW (L-) and OPN1MW (M-)cone opsin genes underlie a spectrum of cone photoreceptor defects from stationary loss of color vision to progressive retinal degeneration. Genotypes of 22 families with a range of cone disorders were grouped into three classes: deletions of the locus control region (LCR); missense mutation (p.Cys203Arg) in an L-/M-hybrid gene; and exon 3 single-nucleotide polymorphism (SNP) interchange haplotypes in an otherwise normal gene array. Moderate-to-high myopia was observed in all mutation categories. Individuals with LCR deletions or p.Cys203Arg mutations were more likely to have nystagmus and poor vision, with disease progression in some p.Cys203Arg patients. Three disease-associated exon 3 SNP haplotypes encoding LIAVA, LVAVA, or MIAVA were identified in our cohort. These patients were less likely to have nystagmus but more likely to show progression, with all patients over the age of 40 years having marked macular abnormalities. Previously, the haplotype LIAVA has been shown to result in exon 3 skipping. Here, we show that haplotypes LVAVA and MIAVA also result in aberrant splicing, with a residual low level of correctly spliced cone opsin. The OPN1LW/OPN1MW:c.532A>G SNP, common to all three disease-associated haplotypes, appears to be principally responsible for this mutational mechanism. PMID:25168334

  6. Parapinopsin, a novel catfish opsin localized to the parapineal organ, defines a new gene family.

    PubMed

    Blackshaw, S; Snyder, S H

    1997-11-01

    Multiple sites of extraretinal photoreception are present in vertebrates, but the molecular basis of extraretinal phototransduction is poorly understood. This study reports the cloning of the first opsin specifically expressed in the directly photosensitive pineal and parapineal of cold-blooded vertebrates. This opsin, identified in channel catfish and termed parapinopsin, defines a new gene family of vertebrate photopigments and is expressed in a majority of parapinealocytes and a subset of pineal photoreceptor cells. Parapinopsin shows a caudal-rostral gradient of expression within the pineal organ. This study also reports the cloning of partial cDNAs encoding the channel catfish orthologues of rhodopsin and the red cone pigment-the full complement of retinal opsins in the species. In situ hybridization studies using probes derived from these retinal opsins, together with parapinopsin, reveal no expression of retinal opsins in pineal and parapineal organ and no expression of any opsin tested in the "deep brain," iris, or dermal melanophores. These data imply that phototransduction in these sites of extraretinal photoreception must be mediated by novel opsins. PMID:9334384

  7. Spectral tuning by opsin coexpression in retinal regions that view different parts of the visual field.

    PubMed

    Dalton, Brian E; Loew, Ellis R; Cronin, Thomas W; Carleton, Karen L

    2014-12-22

    Vision frequently mediates critical behaviours, and photoreceptors must respond to the light available to accomplish these tasks. Most photoreceptors are thought to contain a single visual pigment, an opsin protein bound to a chromophore, which together determine spectral sensitivity. Mechanisms of spectral tuning include altering the opsin, changing the chromophore and incorporating pre-receptor filtering. A few exceptions to the use of a single visual pigment have been documented in which a single mature photoreceptor coexpresses opsins that form spectrally distinct visual pigments, and in these exceptions the functional significance of coexpression is unclear. Here we document for the first time photoreceptors coexpressing spectrally distinct opsin genes in a manner that tunes sensitivity to the light environment. Photoreceptors of the cichlid fish, Metriaclima zebra, mix different pairs of opsins in retinal regions that view distinct backgrounds. The mixing of visual pigments increases absorbance of the corresponding background, potentially aiding the detection of dark objects. Thus, opsin coexpression may be a novel mechanism of spectral tuning that could be useful for detecting prey, predators and mates. However, our calculations show that coexpression of some opsins can hinder colour discrimination, creating a trade-off between visual functions. PMID:25377457

  8. Spectral tuning by opsin coexpression in retinal regions that view different parts of the visual field

    PubMed Central

    Dalton, Brian E.; Loew, Ellis R.; Cronin, Thomas W.; Carleton, Karen L.

    2014-01-01

    Vision frequently mediates critical behaviours, and photoreceptors must respond to the light available to accomplish these tasks. Most photoreceptors are thought to contain a single visual pigment, an opsin protein bound to a chromophore, which together determine spectral sensitivity. Mechanisms of spectral tuning include altering the opsin, changing the chromophore and incorporating pre-receptor filtering. A few exceptions to the use of a single visual pigment have been documented in which a single mature photoreceptor coexpresses opsins that form spectrally distinct visual pigments, and in these exceptions the functional significance of coexpression is unclear. Here we document for the first time photoreceptors coexpressing spectrally distinct opsin genes in a manner that tunes sensitivity to the light environment. Photoreceptors of the cichlid fish, Metriaclima zebra, mix different pairs of opsins in retinal regions that view distinct backgrounds. The mixing of visual pigments increases absorbance of the corresponding background, potentially aiding the detection of dark objects. Thus, opsin coexpression may be a novel mechanism of spectral tuning that could be useful for detecting prey, predators and mates. However, our calculations show that coexpression of some opsins can hinder colour discrimination, creating a trade-off between visual functions. PMID:25377457

  9. Molecular expression of opsin gene in growing juvenile mackerel ( Scomber japonicus Houttuyn)

    NASA Astrophysics Data System (ADS)

    Kim, Eung-Oh; Yoon, Seong-Jong; Park, Kyoung-Hyun; Kim, Dae-Hyun; Do, Jeung-Wan; Cho, Eun-Seob

    2009-12-01

    Fish have developed color vision that is closely adapted to their photic environments, where both spectral sensitivity and the number of visual opsins are influenced. The mackerel used in this study is one of the most important fishery stocks in Korea. The opsin gene of the mackerel juveniles after 20 days in hatching was isolated and characterized based on the molecular study of visual photoreceptor. The full-length mackerel opsin gene was obtained by PCR amplification of genomic DNA, as well as cDNA synthesis. Sequence analysis of the opsin gene showed that it contained a 1,080 bp open reading frame encoding 360 amino acids. Based on Schiff’s base formation (S114, K119), glycosylation (E3, F37) and palmitoylation (S281, 282), the deduced amino acid sequence had a typical rod opsin. The mackerel and Gempylus serpens showed 73.7% DNA homology on opsin gene, which was higher than any other of investigated species. In the analysis of phylogenetic relationship, the genetic placement of the mackerel is closer to that of Scombroidei than Labroidei, with supporting somewhat strong bootstrap value. In the analysis of Northern and RT-PCR, the probed products were observed only in rapidly growing juveniles. These findings indicate that in mackerel opsin mRNA expression can be detected in day-20 hatching larvae. It may play an important role in stimulating growth hormone.

  10. Vertebrate ancient (VA) opsin and extraretinal photoreception in the Atlantic salmon (Salmo salar).

    PubMed

    Philp, A R; Garcia-Fernandez, J M; Soni, B G; Lucas, R J; Bellingham, J; Foster, R G

    2000-06-01

    A member of a new photopigment family first isolated from teleost fish, vertebrate ancient (VA) opsin, has recently been shown to form a functional photopigment and to be expressed within a subset of horizontal and amacrine cells of the inner retina. These sites of expression (and structural features) of VA opsin suggest that this photopigment might mediate non-image-forming light-detection tasks. We attempted to gain support for this hypothesis by examining the expression of VA opsin within the central nervous system (CNS) (pineal and deep brain) of the Atlantic salmon Salmo salar. In addition, we examined the sites of rod-opsin, cone-opsin and &agr; -transducin expression within the salmon CNS to provide a more complete description of the extraretinal photoreceptors of a teleost vertebrate. We show that multiple populations of cells within the salmon CNS appear to contain photoreceptors: VA opsin was strongly expressed in the pineal organ and in bilateral columns of subependymal cells in the epithalamus; anti-cone-opsin antibodies labelled cells within the pineal and numerous cells in the anterior hypothalamus (suprachiasmatic nucleus, nucleus preopticus magnocellularis, nucleus preopticus parvocellularis); anti-rod-opsin antibodies labelled cells within the pineal but no other areas within the central brain; and anti- &agr; -transducin antibodies labelled cells within the pineal and the ventral telencephalon. Collectively, our results suggest that VA opsin is a photopigment specialised for irradiance detection tasks within the eye, pineal and central brain, and that the salmon has multiple and varied populations of photoreceptors within the CNS. We review the significance of these findings within the broad context of vertebrate extraretinal photoreception. PMID:10821749

  11. Correlative Sun-Earth-Connections Science Services in the Early Phase of LWS

    NASA Astrophysics Data System (ADS)

    McGuire, R. E.; Burley, R. J.; Candey, R. M.; Chimiak, R. A.; Fung, S. F.; Harris, B. T.; Hills, H. K.; Johnson, R. C.; Kessel, R. L.; Kovalick, T. J.; Leckner, H. A.

    2002-05-01

    The Coordinated Data Analysis [Workshop] Web (CDAWeb) supports graphics and data retrieval from a unique database of current multi-mission Sun-Earth-Connections (SEC) data. The Satellite Situation Center Web (SSCWeb) serves satellite orbits and various multi-mission conjunctions queries. CDAWeb now acts as the US Cluster Science Data Center to serve Cluster Prime Parameters to authorized Cluster science investigators. These systems make current data quickly accessible to a wide research community and immediately available in the context of simultaneous data from almost all other current space physics missions. These systems are among the first working prototypes for delivering the class of integrated, multi-mission data view essential to the programs like Living with a Star (LWS). These systems' databases, capabilities, and usage continue to grow. CDAWeb served 160,000 plots and listings plus 206,000 file downloads in the year 2001, SSCWeb served 39,000 listings, query results and plots. CDAWeb now has 6 years where data from more 100 data sets is available for every day. Over the last year, CDAWeb has been extended to ~500 GBytes and added new datasets from Wind, Polar and other missions. CDAWeb remains hosted at three mirror sites (in Germany-MPE, England-RAL and Japan-ISAS) as well as by the primary site at Goddard. New higher-resolution data, new direct project data flows, new tools and our plans for evolution will be shown and demonstrated. Supported by the NASA Office of Space Science, CDAWeb and SSCWeb are joint efforts of the NASA GSFC Space Physics Data Facility (SPDF) and the National Space Science Data Center (NSSDC). These services are only possible in turn by data flows from various project data facilities and the work of the instrument science teams on the many missions now supported.

  12. A Ligand Channel through the G Protein Coupled Receptor Opsin

    PubMed Central

    Park, Jung Hee; Choe, Hui-Woog; Piechnick, Ronny; Ernst, Oliver P.; Hofmann, Klaus Peter; Heck, Martin

    2009-01-01

    The G protein coupled receptor rhodopsin contains a pocket within its seven-transmembrane helix (TM) structure, which bears the inactivating 11-cis-retinal bound by a protonated Schiff-base to Lys296 in TM7. Light-induced 11-cis-/all-trans-isomerization leads to the Schiff-base deprotonated active Meta II intermediate. With Meta II decay, the Schiff-base bond is hydrolyzed, all-trans-retinal is released from the pocket, and the apoprotein opsin reloaded with new 11-cis-retinal. The crystal structure of opsin in its active Ops* conformation provides the basis for computational modeling of retinal release and uptake. The ligand-free 7TM bundle of opsin opens into the hydrophobic membrane layer through openings A (between TM1 and 7), and B (between TM5 and 6), respectively. Using skeleton search and molecular docking, we find a continuous channel through the protein that connects these two openings and comprises in its central part the retinal binding pocket. The channel traverses the receptor over a distance of ca. 70 Å and is between 11.6 and 3.2 Å wide. Both openings are lined with aromatic residues, while the central part is highly polar. Four constrictions within the channel are so narrow that they must stretch to allow passage of the retinal β-ionone-ring. Constrictions are at openings A and B, respectively, and at Trp265 and Lys296 within the retinal pocket. The lysine enforces a 90° elbow-like kink in the channel which limits retinal passage. With a favorable Lys side chain conformation, 11-cis-retinal can take the turn, whereas passage of the all-trans isomer would require more global conformational changes. We discuss possible scenarios for the uptake of 11-cis- and release of all-trans-retinal. If the uptake gate of 11-cis-retinal is assigned to opening B, all-trans is likely to leave through the same gate. The unidirectional passage proposed previously requires uptake of 11-cis-retinal through A and release of photolyzed all-trans-retinal through B. PMID:19194506

  13. Optogenetics: 10 years of microbial opsins in neuroscience

    PubMed Central

    Deisseroth, Karl

    2016-01-01

    Over the past 10 years, the development and convergence of microbial opsin engineering, modular genetic methods for cell-type targeting and optical strategies for guiding light through tissue have enabled versatile optical control of defined cells in living systems, defining modern optogenetics. Despite widespread recognition of the importance of spatiotemporally precise causal control over cellular signaling, for nearly the first half (2005–2009) of this 10-year period, as optogenetics was being created, there were difficulties in implementation, few publications and limited biological findings. In contrast, the ensuing years have witnessed a substantial acceleration in the application domain, with the publication of thousands of discoveries and insights into the function of nervous systems and beyond. This Historical Commentary reflects on the scientific landscape of this decade-long transition. PMID:26308982

  14. The amino terminus of opsin translocates "posttranslationally" as efficiently as cotranslationally.

    PubMed

    Kanner, Elliott M; Klein, Irene K; Friedlander, Martin; Simon, Sanford M

    2002-06-18

    Opsin, a member of the G-protein-coupled receptor family, is a polytopic membrane protein that does not encode a cleaved amino-terminal signal sequence. The amino terminus of opsin precedes the first known targeting information, suggesting that it translocates across the endoplasmic reticulum (ER) membrane after synthesis, uncoupled from translation. However, translocation across the mammalian ER is believed to be coupled to protein synthesis. In this study we show that opsin, within a range of nascent peptide lengths, targets and translocates equally efficiently co- and posttranslationally. Longer nascent opsin peptides have a lower efficiency of cotranslational translocation but an even lower efficiency of posttranslational translocation. We also show that SRP is required for both co- and posttranslational targeting. PMID:12056902

  15. The photochemical determinants of color vision: revealing how opsins tune their chromophore's absorption wavelength.

    PubMed

    Wang, Wenjing; Geiger, James H; Borhan, Babak

    2014-01-01

    The evolution of a variety of important chromophore-dependent biological processes, including microbial light sensing and mammalian color vision, relies on protein modifications that alter the spectral characteristics of a bound chromophore. Three different color opsins share the same chromophore, but have three distinct absorptions that together cover the entire visible spectrum, giving rise to trichromatic vision. The influence of opsins on the absorbance of the chromophore has been studied through methods such as model compounds, opsin mutagenesis, and computational modeling. The recent development of rhodopsin mimic that uses small soluble proteins to recapitulate the binding and wavelength tuning of the native opsins provides a new platform for studying protein-regulated spectral tuning. The ability to achieve far-red shifted absorption in the rhodopsin mimic system was attributed to a combination of the lack of a counteranion proximal to the iminium, and a uniformly neutral electrostatic environment surrounding the chromophore. PMID:24323922

  16. Broadband activation by white-opsin lowers intensity threshold for cellular stimulation

    PubMed Central

    Batabyal, Subrata; Cervenka, Gregory; Birch, David; Kim, Young-tae; Mohanty, Samarendra

    2015-01-01

    Photoreceptors, which initiate the conversion of ambient light to action potentials via retinal circuitry, degenerate in retinal diseases such as retinitis pigmentosa and age related macular degeneration leading to loss of vision. Current prosthetic devices using arrays consisting of electrodes or LEDs (for optogenetic activation of conventional narrow-band opsins) have limited spatial resolution and can cause damage to retinal circuits by mechanical or photochemical (by absorption of intense narrow band light) means. Here, we describe a broad-band light activatable white-opsin for generating significant photocurrent at white light intensity levels close to ambient daylight conditions. White-opsin produced an order of magnitude higher photocurrent in response to white light as compared to narrow-band opsin channelrhodopsin-2, while maintaining the ms-channel kinetics. High fidelity of peak-photocurrent (both amplitude and latency) of white-opsin in response to repetitive white light stimulation of varying pulse width was observed. The significantly lower intensity stimulation required for activating white-opsin sensitized cells may facilitate ambient white light-based restoration of vision for patients with widespread photoreceptor degeneration. PMID:26658483

  17. Chromatic Properties of Horizontal and Ganglion Cell Responses Follow a Dual Gradient in Cone Opsin Expression

    PubMed Central

    Yin, Lu; Smith, Robert G; Sterling, Peter; Brainard, David H.

    2007-01-01

    In guinea pig retina, immunostaining reveals a dual gradient of opsins: cones expressing opsin sensitive to medium wavelengths (M) predominate in the upper retina, whereas cones expressing opsin sensitive to shorter wavelengths (S) predominate in the lower retina. Whether these gradients correspond to functional gradients in postreceptoral neurons is essentially unknown. Using monochromatic flashes, we measured the relative weights with which M, S, and rod signals contribute to horizontal cell responses. For a background that produced 4.76 log10 photoisomerizations per rod per second (Rh*/rod/s), mean weights in superior retina were 52% (M), 2% (S), and 46% (rod). Mean weights in inferior retina were 9% (M), 50% (S), and 41% (rod). In superior retina, cone opsin weights agreed quantitatively with relative pigment density estimates from immunostaining. In inferior retina, cone opsin weights agreed qualitatively with relative pigment density estimates, but quantitative comparison was impossible because individual cones coexpress both opsins to varying and unquantifiable degrees. We further characterized the functional gradients in horizontal and brisk-transient ganglion cells using flickering stimuli produced by various mixtures of blue and green primary lights. Cone weights for both cell types resembled those obtained for horizontal cells using monochromatic flashes. Because the brisk-transient ganglion cell is thought to mediate behavioral detection of luminance contrast, our results are consistent with the hypothesis that the dual gradient of cone opsins assists achromatic contrast detection against different spectral backgrounds. In our preparation, rod responses did not completely saturate, even at background light levels typical of outdoor sunlight (5.14 log10 Rh*/rod/s). PMID:17122060

  18. X-Linked Cone Dystrophy Caused by Mutation of the Red and Green Cone Opsins

    PubMed Central

    Gardner, Jessica C.; Webb, Tom R.; Kanuga, Naheed; Robson, Anthony G.; Holder, Graham E.; Stockman, Andrew; Ripamonti, Caterina; Ebenezer, Neil D.; Ogun, Olufunmilola; Devery, Sophie; Wright, Genevieve A.; Maher, Eamonn R.; Cheetham, Michael E.; Moore, Anthony T.; Michaelides, Michel; Hardcastle, Alison J.

    2010-01-01

    X-linked cone and cone-rod dystrophies (XLCOD and XLCORD) are a heterogeneous group of progressive disorders that solely or primarily affect cone photoreceptors. Mutations in exon ORF15 of the RPGR gene are the most common underlying cause. In a previous study, we excluded RPGR exon ORF15 in some families with XLCOD. Here, we report genetic mapping of XLCOD to Xq26.1-qter. Asignificant LOD score was detected with marker DXS8045 (Zmax = 2.41 [? = 0.0]). The disease locus encompasses the cone opsin gene array on Xq28. Analysis of the array revealed a missense mutation (c. 529T>C [p. W177R]) in exon 3 of both the long-wavelength-sensitive (LW, red) and medium-wavelength-sensitive (MW, green) cone opsin genes that segregated with disease. Both exon 3 sequences were identical and were derived from the MW gene as a result of gene conversion. The amino acid W177 is highly conserved in visual and nonvisual opsins across species. We show that W177R in MW opsin and the equivalent W161R mutation in rod opsin result in protein misfolding and retention in the endoplasmic reticulum. We also demonstrate that W177R misfolding, unlike the P23H mutation in rod opsin that causes retinitis pigmentosa, is not rescued by treatment with the pharmacological chaperone 9-cis-retinal. Mutations in the LW/MW cone opsin gene array can, therefore, lead to a spectrum of disease, ranging from color blindness to progressive cone dystrophy (XLCOD5). PMID:20579627

  19. Brain area-specific diurnal and photic regulation of val-opsinA and val-opsinB genes in the zebrafish.

    PubMed

    Hang, Chong Yee; Kitahashi, Takashi; Parhar, Ishwar S

    2015-05-01

    Zebrafish possess two isoforms of vertebrate ancient long (VAL)-opsin, val-opsinA (valopa) and val-opsinB (valopb), which probably mediate non-visual responses to light. To understand the diurnal and light-sensitive regulation of the valop genes in different cell groups, the current study used real-time quantitative PCR to examine the diurnal changes of valopa and b mRNA levels in different brain areas of adult male zebrafish. Furthermore, effects of the extended exposure to light or dark condition, luminous levels and the treatment with a melatonin receptor agonist or antagonist on valop transcription were examined. In the thalamus, valop mRNA levels showed significant diurnal changes; valopa peaked in the evening, while valopb peaked in the morning. The diurnal change of valopa mRNA levels occurred independent of light conditions, whereas that of valopb mRNA levels were regulated by light. A melatonin receptor agonist or antagonist did not affect the changes of valop mRNA levels. In contrast, the midbrain and hindbrain showed arrhythmic valop mRNA levels under light and dark cycles. The differential diurnal regulation of the valopa and b genes in the thalamus and the arrhythmic expression in the midbrain and hindbrain suggest involvement of deep brain VAL-opsin in time- and light-dependent physiology. We show diurnal expression changes of vertebrate ancient long (VAL) opsin genes (valopa and valopb), depending on brain area, time of day and light condition, in the adult male zebrafish. Differential regulation of the valop genes in the thalamus and arrhythmic expression in the midbrain and hindbrain suggest their involvement in time- and light-dependent physiology to adjust to environmental changes. PMID:25727787

  20. Spectral sensitivity in Onychophora (velvet worms) revealed by electroretinograms, phototactic behaviour and opsin gene expression.

    PubMed

    Beckmann, Holger; Hering, Lars; Henze, Miriam J; Kelber, Almut; Stevenson, Paul A; Mayer, Georg

    2015-03-01

    Onychophorans typically possess a pair of simple eyes, inherited from the last common ancestor of Panarthropoda (Onychophora+Tardigrada+Arthropoda). These visual organs are thought to be homologous to the arthropod median ocelli, whereas the compound eyes probably evolved in the arthropod lineage. To gain insights into the ancestral function and evolution of the visual system in panarthropods, we investigated phototactic behaviour, opsin gene expression and the spectral sensitivity of the eyes in two representative species of Onychophora: Euperipatoides rowelli (Peripatopsidae) and Principapillatus hitoyensis (Peripatidae). Our behavioural analyses, in conjunction with previous data, demonstrate that both species exhibit photonegative responses to wavelengths ranging from ultraviolet to green light (370-530 nm), and electroretinograms reveal that the onychophoran eye is maximally sensitive to blue light (peak sensitivity ∼480 nm). Template fits to these sensitivities suggest that the onychophoran eye is monochromatic. To clarify which type of opsin the single visual pigment is based on, we localised the corresponding mRNA in the onychophoran eye and brain using in situ hybridization. Our data show that the r-opsin gene (onychopsin) is expressed exclusively in the photoreceptor cells of the eye, whereas c-opsin mRNA is confined to the optic ganglion cells and the brain. Together, our findings suggest that the onychopsin is involved in vision, whereas c-opsin might have a photoreceptive, non-visual function in onychophorans. PMID:25617459

  1. A spinal opsin controls early neural activity and drives a behavioral light response

    PubMed Central

    Friedmann, Drew; Hoagland, Adam; Berlin, Shai; Isacoff, Ehud Y.

    2014-01-01

    Non-visual detection of light by the vertebrate hypothalamus, pineal, and retina is known to govern seasonal and circadian behaviors [1]. However, the expression of opsins in multiple other brain structures [2–4] suggests a more expansive repertoire for light-regulation of physiology, behavior, and development. Translucent zebrafish embryos express extra-retinal opsins early on [5, 6], at a time when spontaneous activity in the developing central nervous system plays a role in neuronal maturation and circuit formation [7]. Though the presence of extra-retinal opsins is well documented, the function of direct photoreception by the central nervous system remains largely unknown. Here we show that early activity in the zebrafish spinal central pattern generator (CPG) and the earliest locomotory behavior are dramatically inhibited by physiological levels of environmental light. We find that the photo-sensitivity of this circuit is conferred by vertebrate ancient long opsin (VALopA), which we show to be a Gαi-coupled receptor that is expressed in the neurons of the spinal network. Sustained photo-activation of VALopA not only suppresses spontaneous activity but also alters the maturation of time-locked correlated network patterns. These results uncover a novel role for non-visual opsins and a mechanism for environmental regulation of spontaneous motor behavior and neural activity in a circuit previously thought to be governed only by intrinsic developmental programs. PMID:25484291

  2. A spinal opsin controls early neural activity and drives a behavioral light response.

    PubMed

    Friedmann, Drew; Hoagland, Adam; Berlin, Shai; Isacoff, Ehud Y

    2015-01-01

    Nonvisual detection of light by the vertebrate hypothalamus, pineal, and retina is known to govern seasonal and circadian behaviors. However, the expression of opsins in multiple other brain structures suggests a more expansive repertoire for light regulation of physiology, behavior, and development. Translucent zebrafish embryos express extraretinal opsins early on, at a time when spontaneous activity in the developing CNS plays a role in neuronal maturation and circuit formation. Though the presence of extraretinal opsins is well documented, the function of direct photoreception by the CNS remains largely unknown. Here, we show that early activity in the zebrafish spinal central pattern generator (CPG) and the earliest locomotory behavior are dramatically inhibited by physiological levels of environmental light. We find that the photosensitivity of this circuit is conferred by vertebrate ancient long opsin A (VALopA), which we show to be a Gα(i)-coupled receptor that is expressed in the neurons of the spinal network. Sustained photoactivation of VALopA not only suppresses spontaneous activity but also alters the maturation of time-locked correlated network patterns. These results uncover a novel role for nonvisual opsins and a mechanism for environmental regulation of spontaneous motor behavior and neural activity in a circuit previously thought to be governed only by intrinsic developmental programs. PMID:25484291

  3. Crepuscular Behavioral Variation and Profiling of Opsin Genes in Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Jenkins, Adam M; Muskavitch, Marc A T

    2015-05-01

    We understand little about photo-preference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photo-preference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photo-preference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photo-preference behavior (e.g., visual sensitivity). PMID:26334802

  4. Crepuscular Behavioral Variation and Profiling of Opsin Genes in Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae)

    PubMed Central

    Jenkins, Adam M.; Muskavitch, Marc A. T.

    2015-01-01

    We understand little about photopreference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photopreference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photopreference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photopreference behavior (e.g., visual sensitivity). PMID:26334802

  5. The hypothalamic photoreceptors regulating seasonal reproduction in birds: a prime role for VA opsin.

    PubMed

    Garca-Fernndez, Jos M; Cernuda-Cernuda, Rafael; Davies, Wayne I L; Rodgers, Jessica; Turton, Michael; Peirson, Stuart N; Follett, Brian K; Halford, Stephanie; Hughes, Steven; Hankins, Mark W; Foster, Russell G

    2015-04-01

    Extraretinal photoreceptors located within the medio-basal hypothalamus regulate the photoperiodic control of seasonal reproduction in birds. An action spectrum for this response describes an opsin photopigment with a ?max of ? 492 nm. Beyond this however, the specific identity of the photopigment remains unresolved. Several candidates have emerged including rod-opsin; melanopsin (OPN4); neuropsin (OPN5); and vertebrate ancient (VA) opsin. These contenders are evaluated against key criteria used routinely in photobiology to link orphan photopigments to specific biological responses. To date, only VA opsin can easily satisfy all criteria and we propose that this photopigment represents the prime candidate for encoding daylength and driving seasonal breeding in birds. We also show that VA opsin is co-expressed with both gonadotropin-releasing hormone (GnRH) and arginine-vasotocin (AVT) neurons. These new data suggest that GnRH and AVT neurosecretory pathways are endogenously photosensitive and that our current understanding of how these systems are regulated will require substantial revision. PMID:25448788

  6. Regeneration of bovine and octopus opsins in situ with natural and artificial retinals

    SciTech Connect

    Koutalos, Y.; Ebrey, T.G.; Tsuda, M.; Odashima, K.; Lien, T.; Park, M.H.; Shimizu, N.; Derguini, F.; Nakanishi, K.; Gilson, H.R.; Honig, B. )

    1989-03-21

    The authors consider the problem of color regulation in visual pigments for both bovine rhodopsin and octopus rhodopsin. Both pigments have 11-cis-retinal as their chromophore. These rhodopsins were bleached in their native membranes, and the opsins were regenerated with natural and artificial chromophores. Both bovine and octopus opsins were regenerated with the 9-cis- and 11-cis-retinal isomers, but the octopus opsin was additionally regenerated with the 13-cis and all-trans isomers. Titration of the octopus opsin with 11-cis-retinal gave an extinction coefficient for octopus rhodopsin of 27,000 {plus minus} 3,000 M{sup {minus}1} cm{sup {minus}1} at 475 nm. The absorption maxima of bovine artificial pigments formed by regenerating opsin with the 11-cis dihydro series of chromophores support a color regulation model for bovine rhodopsin in which the chromophore-binding site of the protein has two negative charges: one directly hydrogen bonded to the Schiff base nitrogen and another near carbon-13. Formation of octopus artificial pigments with both all-trans and 11-cis dihydro chromophores leads to a similar model for octopus rhodopsin and metarhodopsin: there are two negative charges in the chromophore-binding site, one directly hydrogen bonded to the Schiff base nitrogen and a second near carbon-13. The interaction of this second charge with the chromophore in octopus rhodopsin is weaker than in bovine, while in metarhodopsin it is as strong as in bovine.

  7. Broad-band opsin for effective stimulation of cells by white light

    NASA Astrophysics Data System (ADS)

    Batabyal, Subrata; Cervenka, Gregory; Kim, Young-Tae; Mohanty, Samarendra

    2015-03-01

    Currently, use of optogenetic sensitization of retinal cells combined with activation/inhibition has potential as alternative to retinal implants that would have required electrodes inside every single neuron for high visual resolution. However, clinical translation of optogenetic activation for restoration of vision suffers from the drawback that narrow spectral sensitivity of opsin requires active stimulation by blue laser or LED having intensity much higher than ambient light. In order to allow ambient-light based stimulation paradigm, here we report development of broad-band opsin that has broad spectral excitability in the entire visible spectrum. The cells sensitized with the broad-band opsin showed order of magnitude higher excitability with white light as compared to that using only the narrow-band light components. The use of broad-band opsin construct will allow higher sensitivity of the opsin-sensitized neurons in degenerated retina to ambient white light, and therefore, significantly lower activation-threshold in contrast to conventional approach of intense, narrow-band light based active-stimulation.

  8. The Verriest Lecture: Short-wave-sensitive cone pathways across the life span.

    PubMed

    Werner, John S

    2016-03-01

    Structurally and functionally, the short-wave-sensitive (S) cone pathways are thought to decline more rapidly with normal aging than the middle- and long-wave-sensitive cone pathways. This would explain the celebrated results by Verriest and others demonstrating that the largest age-related color discrimination losses occur for stimuli on a tritan axis. Here, we challenge convention, arguing from psychophysical data that selective S-cone pathway losses do not cause declines in color discrimination. We show substantial declines in chromatic detection and discrimination, as well as in temporal and spatial vision tasks, that are mediated by S-cone pathways. These functional losses are not, however, unique to S-cone pathways. Finally, despite reduced photon capture by S cones, their postreceptoral pathways provide robust signals for the visual system to renormalize itself to maintain nearly stable color perception across the life span. PMID:26974914

  9. Opsin expression in Limulus eyes: a UV opsin is expressed in each eye type and co-expressed with a visible light-sensitive opsin in ventral larval eyes

    PubMed Central

    Battelle, Barbara-Anne; Kempler, Karen E.; Harrison, Alexandra; Dugger, Donald R.; Payne, Richard

    2014-01-01

    The eyes of the horseshoe crab, Limulus polyphemus, are a model for studies of visual function and the visual systems of euarthropods. Much is known about the structure and function of L. polyphemus photoreceptors, much less about their photopigments. Three visible-light-sensitive L. polyphemus opsins were characterized previously (LpOps1, 2 and 5). Here we characterize a UV opsin (LpUVOps1) that is expressed in all three types of L. polyphemus eyes. It is expressed in most photoreceptors in median ocelli, the only L. polyphemus eyes in which UV sensitivity was previously detected, and in the dendrite of eccentric cells in lateral compound eyes. Therefore, eccentric cells, previously thought to be non-photosensitive second-order neurons, may actually be UV-sensitive photoreceptors. LpUVOps1 is also expressed in small photoreceptors in L. polyphemus ventral larval eyes, and intracellular recordings from these photoreceptors confirm that LpUVOps1 is an active, UV-sensitive photopigment. These photoreceptors also express LpOps5, which we demonstrate is an active, long-wavelength-sensitive photopigment. Thus small photoreceptors in ventral larval eyes, and probably those of the other larval eyes, have dual sensitivity to UV and visible light. Interestingly, the spectral tuning of small ventral photoreceptors may change day to night, because the level of LpOps5 in their rhabdoms is lower during the day than during the night, whereas LpUVOps1 levels show no diurnal change. These and previous findings show that opsin co-expression and the differential regulation of co-expressed opsins in rhabdoms is a common feature of L. polyphemus photoreceptors. PMID:24948643

  10. Adaptive evolution of cone opsin genes in two colorful cyprinids, Opsariichthys pachycephalus and Candidia barbatus.

    PubMed

    Wang, Feng Yu; Chung, Wen Sung; Yan, Hong Young; Tzeng, Chyng Shyan

    2008-07-01

    Opsariichthys pachycephalus and Candidia barbatus are two phylogenetically related freshwater cyprinids that both exhibit colorful, yet quite different nuptial coloration. This study was designed to test the hypothesis that differences in nuptial coloration between two species could reflect differences in color perception ability and the opsin genes that coded for it. Genes encoding the visual pigments of these two species were cloned and sequenced, lambda(max) of cone photoreceptors and the reflectance spectra of their body coloration were measured to test the hypothesis. The 14-nm spectral shift between green-light-sensitive photoreceptors of these two cyprinids is found to correlate well with differences in their reflective spectra. The spectral shift could result from differential expression of opsin genes and the interactive effects of the amino acid replacements in various minor sites. These results support our hypothesis that nuptial coloration is tied to color perception ability and opsin genes. PMID:18571688

  11. The opsins of the vertebrate retina: insights from structural, biochemical, and evolutionary studies.

    PubMed

    Nickle, B; Robinson, P R

    2007-11-01

    The vertebrate retina contains several classes of visual pigments responsible for such diverse functions as image- and nonimage-forming vision, the entrainment of circadian cycles, and the pupilary light response. With vision being vital to the survival of many species, the elucidation of the structural and biochemical properties of visual pigments has been the focus of a large body of research that has led to rapid advances in the field of photoreception. In this review, the current understanding of the structure, function, biochemistry, and evolution of the opsins that make up the photopigments in the vertebrate retina will be reviewed. These include the rod and cone opsins, melanopsin, RGR, peropsin, and VA-opsin. The goal is to highlight important questions that have been answered and to define some of the remaining questions in the field that will provide future directions for research. PMID:17726575

  12. The Microbial Opsin Homolog Sop1 is involved in Sclerotinia sclerotiorum Development and Environmental Stress Response

    PubMed Central

    Lyu, Xueliang; Shen, Cuicui; Fu, Yanping; Xie, Jiatao; Jiang, Daohong; Li, Guoqing; Cheng, Jiasen

    2016-01-01

    Microbial opsins play a crucial role in responses to various environmental signals. Here, we report that the microbial opsin homolog gene sop1 from the necrotrophic phytopathogenic fungus Sclerotinia sclerotiorum was dramatically up-regulated during infection and sclerotial development compared with the vegetative growth stage. Further, study showed that sop1 was essential for growth, sclerotial development and full virulence of S. sclerotiorum. Sop1-silenced transformants were more sensitive to high salt stress, fungicides and high osmotic stress. However, they were more tolerant to oxidative stress compared with the wild-type strain, suggesting that sop1 is involved in different stress responses and fungicide resistance, which plays a role in the environmental adaptability of S. sclerotiorum. Furthermore, a Delta blast search showed that microbial opsins are absent from the genomes of animals and most higher plants, indicating that sop1 is a potential drug target for disease control of S. sclerotiorum. PMID:26779159

  13. Contributions of rhodopsin, cone opsins, and melanopsin to postreceptoral pathways inferred from natural image statistics

    PubMed Central

    Barrionuevo, Pablo A.; Cao, Dingcai

    2014-01-01

    Visual neural representation is constrained by the statistical properties of the environment. Prior analysis of cone pigment excitations for natural images revealed three principal components corresponding to the major retinogeniculate pathways identified by anatomical and physiological studies in primates. Here, principal component analyses were conducted on the excitations of rhodopsin, cone opsins, and melanopsin for nine hyperspectral images under 21 natural illuminants. The results suggested that rhodopsin and melanopsin may contribute to the three major retinogeniculate pathways. Rhodopsin and melanopsin may provide additional constraints in natural scene statistics, leading to new components that cannot be revealed by analysis based on cone opsin excitations only. PMID:24695161

  14. Vertebrate ancient-long opsin: a green-sensitive photoreceptive molecule present in zebrafish deep brain and retinal horizontal cells.

    PubMed

    Kojima, D; Mano, H; Fukada, Y

    2000-04-15

    Nonretinal/nonpineal photosensitivity has been found in the brain of vertebrates, but the molecular basis for such a "deep brain" photoreception system remains unclear. We conducted an extensive search for brain opsin cDNAs of the zebrafish (Danio rerio), a useful animal model for genetic studies, and we have isolated a partial cDNA clone encoding an ortholog of vertebrate ancient (VA) opsin, the function of which is unknown. Subsequent characterization revealed the occurrence of two kinds of mRNAs encoding putative splicing variants, VA and VA-Long (VAL) opsin, the latter of which is a novel variant of the former. Both opsins shared a common core sequence in the membrane-spanning domains, but VAL-opsin had a C-terminal tail much longer than that of VA-opsin. Functional reconstitution experiments on the recombinant proteins showed that VAL-opsin with bound 11-cis-retinal is a green-sensitive pigment (lambdamax approximately 500 nm), whereas VA-opsin exhibited no photosensitivity even in the presence of 11-cis-retinal. Immunoreactivity specific to this functionally active VAL-opsin was localized at a limited number of cells surrounding the diencephalic ventricle of central thalamus, and these cells were distributed over approximately 200 micrometer along the rostrocaudal axis. Taken together with the previous study on the locus of the teleost brain photosensitivity (von Frisch K, 1911), it is strongly suggested that the VAL-positive cells in the zebrafish brain represent the deep brain photoreceptors. The VAL-specific immunoreactivity was also detected in a subset of non-GABAergic horizontal cells in the zebrafish retina. The existence of VAL-opsin, a new member of the rhodopsin superfamily, in these tissues may indicate its multiple roles in visual and nonvisual photosensory physiology. PMID:10751436

  15. Molecular Evidence that Only Two Opsin Subfamilies, the Blue Light- (SWS2) and Green Light-Sensitive (RH2), Drive Color Vision in Atlantic Cod (Gadus morhua)

    PubMed Central

    Søviknes, Anne Mette; Drivenes, Øyvind; Helvik, Jon Vidar

    2014-01-01

    Teleosts show a great variety in visual opsin complement, due to both gene duplication and gene loss. The repertoire ranges from one subfamily of visual opsins (scotopic vision) including rod opsin only retinas seen in many deep-sea species to multiple subfamilies of visual opsins in some pelagic species. We have investigated the opsin repertoire of Atlantic cod (Gadus morhua) using information in the recently sequenced cod genome and found that despite cod not being a deep sea species it lacks visual subfamilies sensitive towards the most extreme parts of the light spectra representing UV and red light. Furthermore, we find that Atlantic cod has duplicated paralogs of both blue-sensitive SWS2 and green-sensitive RH2 subfamilies, with members belonging to each subfamily linked in tandem within the genome (two SWS2-, and three RH2A genes, respectively). The presence of multiple cone opsin genes indicates that there have been duplication events in the cod ancestor SWS2 and RH2 opsins producing paralogs that have been retained in Atlantic. Our results are supported by expressional analysis of cone opsins, which further revealed an ontogenetic change in the array of cone opsins expressed. These findings suggest life stage specific programs for opsin regulation which could be linked to habitat changes and available light as the larvae is transformed into an early juvenile. Altogether we provide the first molecular evidence for color vision driven by only two families of cone opsins due to gene loss in a teleost. PMID:25551396

  16. Homeobox transcription factor Six7 governs expression of green opsin genes in zebrafish.

    PubMed

    Ogawa, Yohey; Shiraki, Tomoya; Kojima, Daisuke; Fukada, Yoshitaka

    2015-08-01

    Colour discrimination in vertebrates requires cone photoreceptor cells in the retina, and high-acuity colour vision is endowed by a set of four cone subtypes expressing UV-, blue-, green- and red-sensitive opsins. Previous studies identified transcription factors governing cone photoreceptor development in mice, although loss of blue and green opsin genes in the evolution of mammals make it difficult to understand how high-acuity colour vision was organized during evolution and development. Zebrafish (Danio rerio) represents a valuable vertebrate model for studying colour vision as it retains all the four ancestral vertebrate cone subtypes. Here, by RT-qPCR and in situ hybridization analysis, we found that sine oculis homeobox homolog 7 (six7), a transcription factor widely conserved in ray-finned fish, is expressed predominantly in the cone photoreceptors in zebrafish at both the larval and the adult stages. TAL effector nuclease-based six7 knock-out revealed its roles in expression of green, red and blue cone opsin genes. Most prominently, the six7 deficiency caused a loss of expression of all the green opsins at both the larval and adult stages. six7 is indispensable for the development and/or maintenance of the green cones. PMID:26180064

  17. Distinct functions for IFT140 and IFT20 in opsin transport.

    PubMed Central

    Crouse, Jacquelin A.; Lopes, Vanda S.; SanAgustin, Jovenal T.; Keady, Brian T.; Williams, David S.; Pazour, Gregory J.

    2014-01-01

    In the vertebrate retina, light is detected by the outer segments of photoreceptor rods and cones, which are highly modified cilia. Like other cilia, outer segments have no protein synthetic capacity and depend on proteins made in the cell body for their formation and maintenance. The mechanism of transport into the outer segment is not fully understood but intraflagellar transport (IFT) is thought to be a major mechanism for moving protein from the cell body into the cilium. In the case of photoreceptor cells, the high density of receptors and the disk turnover that occurs daily necessitates much higher rates of transport than would be required in other cilia. In this work, we show that the IFT complex A protein IFT140 is required for development and maintenance of outer segments. In earlier work we found that acute deletion of Ift20 caused opsin to accumulate at the Golgi complex. In this work we find that acute deletion of Ift140 does not cause opsin to accumulate at the Golgi complex but rather it accumulates in the plasma membrane of the inner segments. This work is strong support of a model of opsin transport where IFT20 is involved in the movement from the Golgi complex to the base of the cilium. Then, once at the base, the opsin is carried through the connecting cilium by an IFT complex that includes IFT140. PMID:24619649

  18. Opsin transcripts of predatory diving beetles: a comparison of surface and subterranean photic niches

    PubMed Central

    Tierney, Simon M.; Cooper, Steven J. B.; Saint, Kathleen M.; Bertozzi, Terry; Hyde, Josephine; Humphreys, William F.; Austin, Andrew D.

    2015-01-01

    The regressive evolution of eyes has long intrigued biologists yet the genetic underpinnings remain opaque. A system of discrete aquifers in arid Australia provides a powerful comparative means to explore trait regression at the genomic level. Multiple surface ancestors from two tribes of diving beetles (Dytiscidae) repeatedly invaded these calcrete aquifers and convergently evolved eye-less phenotypes. We use this system to assess transcription of opsin photoreceptor genes among the transcriptomes of two surface and three subterranean dytiscid species and test whether these genes have evolved under neutral predictions. Transcripts for UV, long-wavelength and ciliary-type opsins were identified from the surface beetle transcriptomes. Two subterranean beetles showed parallel loss of all opsin transcription, as expected under ‘neutral’ regressive evolution. The third species Limbodessus palmulaoides retained transcription of a long-wavelength opsin (lwop) orthologue, albeit in an aphotic environment. Tests of selection on lwop indicated no significant differences between transcripts derived from surface and subterranean habitats, with strong evidence for purifying selection acting on L. palmulaoides lwop. Retention of sequence integrity and the lack of evidence for neutral evolution raise the question of whether we have identified a novel pleiotropic role for lwop, or an incipient phase of pseudogene development. PMID:26064586

  19. The lycaenid butterfly Polyommatus icarus uses a duplicated blue opsin to see green.

    PubMed

    Sison-Mangus, Marilou P; Briscoe, Adriana D; Zaccardi, Guillermo; Knüttel, Helge; Kelber, Almut

    2008-02-01

    The functional significance of gene duplication is rarely addressed at the level of animal behavior. Butterflies are excellent models in this regard because they can be trained and the use of their opsin-based visual pigments in color vision can be assessed. In the present study, we demonstrate that the lycaenid Polyommatus icarus uses its duplicate blue (B2) opsin, BRh2, in conjunction with its long-wavelength (LW) opsin, LWRh, to see color in the green part of the light spectrum extending up to 560 nm. This is in contrast to butterflies in the genus Papilio, which use duplicate LW opsins to discriminate colors in the long-wavelength range. We also found that P. icarus has a heterogeneously expressed red filtering pigment and red-reflecting ommatidia in the ventral eye region. In behavioural tests, the butterflies could not discriminate colors in the red range (570-640 nm). This finding is significant because we have previously found that the nymphalid butterfly Heliconius erato has filter-pigment mediated color vision in the long wavelength range. Our results suggest that lateral filtering pigments may not always influence color vision in insects. PMID:18203991

  20. Novel missense mutations in red/green opsin genes in congenital color-vision deficiencies.

    PubMed

    Ueyama, Hisao; Kuwayama, Shigeki; Imai, Hiroo; Tanabe, Shoko; Oda, Sanae; Nishida, Yasuhiro; Wada, Akimori; Shichida, Yoshinori; Yamade, Shinichi

    2002-06-01

    The DNAs from 217 Japanese males with congenital red/green color-vision deficiencies were analyzed. Twenty-three subjects had the normal genotype of a single red gene, followed by a green gene. Four of the 23 were from the 69 protan subject group and 19 of the 23 were from the 148 deutan subject group. Three of the 23 subjects had missense mutations. The mutation Asn94Lys (AAC-->AAA) occurred in the single green gene of a deutan subject (A155). The Arg330Gln (CGA-->CAA) mutation was detected in both green genes of another deutan subject (A164). The Gly338Glu (GGG-->GAG) mutation occurred in the single red gene of a protan subject (A89). Both normal and mutant opsins were expressed in cultured COS-7 cells and visual pigments were regenerated with 11-cis-retinal. The normal red and green opsins showed absorbance spectra with lambda(max) of 560 and 530 nm, respectively, but the three mutant opsins had altered spectra. The mutations in Asn94Lys and Gly338Glu resulted in no absorbance and the Arg330Gln mutation gave a low absorbance spectrum with a lambda(max) of 530 nm. Therefore these three mutant opsins are likely to be affected in the folding process, resulting in a loss of function as a visual pigment. PMID:12051694

  1. Intron splice sites of Papilio glaucus PglRh3 corroborate insect opsin phylogeny.

    PubMed

    Briscoe, A D

    1999-04-01

    Full-length cDNA clones encoding the PglRh3 opsin from the tiger swallowtail butterfly Papilio glaucus were isolated from cDNA synthesized from adult head tissue total RNA. This cDNA consists of 1679 nucleotides and contains a single open reading frame predicted to be 379 amino acids in length. PCR amplification of genomic DNA with primers spanning the coding region yielded a single 2760bp fragment which was sequenced. The PglRh3 gene has nine exons and eight introns, four of which are in unique locations relative to the positions of introns in other known insect opsin sequences. Phylogenetic analyses of amino acid and nucleotide sequence data places PglRh3 within a clade of insect visual pigments thought to be sensitive to long wavelengths of light. The genomic structure of PglRh3 is the first characterized from a member of this opsin clade. Three PglRh3 intron positions are shared with Drosophila Rh1, and one of these is also shared with Drosophila Rh2. By contrast, none of the known intron locations in a clade of anciently diverged ultraviolet- and blue-sensitive visual pigments are shared by P. glaucus PglRh3, Drosophila Rh1 or Rh2. The placement of introns within opsin genes therefore independently supports the clustering of a putatively long-wavelength-sensitive clade with a clade of blue-green-sensitive visual pigments. PMID:10196479

  2. Expression of pineal ultraviolet- and green-like opsins in the pineal organ and retina of teleosts.

    PubMed

    Forsell, J; Ekström, P; Flamarique, I N; Holmqvist, B

    2001-07-01

    In teleostean bony fishes, studies on the adults of various species have shown that pineal photoreceptors are maximally sensitive to short- and middle-wavelength light, possibly utilising both rod-like and pineal-specific opsins. Until recently, however, very little was known about the pineal opsins present in embryonic and larval teleosts and their relationships to opsins expressed by retinal photoreceptors. Our immunocytochemical studies have revealed that, in Atlantic halibut, herring and cod, pineal photoreceptors express principal phototransduction molecules during embryonic life before they appear in retinal photoreceptors. In cDNA from embryonic and adult halibut, we identified two partial opsin gene sequences, HPO1 and HPO4, with highest homology to teleost green and ultraviolet cone opsins (72-83% and 71-83% amino acid identity, respectively). In halibut, these opsins are expressed in the pineal organ of embryos and appear in the retina of larvae. Our recent in situ hybridisation studies with RNA probes for HPO1 and HPO4 demonstrate the presence of green-like opsin mRNAs in the pineal organ and the retina of herring, cod, turbot, haddock, Atlantic salmon, zebrafish and three species of cichlid, and of ultraviolet opsins in the retinas of zebrafish, Atlantic salmon, turbot and the three cichlid species. We conclude that the halibut pineal organ appears to have the potential for both ultraviolet and green photosensitivity from the embryonic stage and that the retina may acquire the same potential during the larval stages. In the other teleosts studied, although both pineal and retinal photoreceptors seem to utilise a green-like opsin from the larval stage, ultraviolet photoreception appears to be restricted to the retina. PMID:11511667

  3. Rod and cone opsin families differ in spectral tuning domains but not signal transducing domains as judged by saturated evolutionary trace analysis.

    PubMed

    Carleton, Karen L; Spady, Tyrone C; Cote, Rick H

    2005-07-01

    The visual receptor of rods and cones is a covalent complex of the apoprotein, opsin, and the light-sensitive chromophore, 11-cis-retinal. This pigment must fulfill many functions including photoactivation, spectral tuning, signal transmission, inactivation, and chromophore regeneration. Rod and cone photoreceptors employ distinct families of opsins. Although it is well known that these opsin families provide unique ranges in spectral sensitivity, it is unclear whether the families have additional functional differences. In this study, we use evolutionary trace (ET) analysis of 188 vertebrate opsin sequences to identify functionally important sites in each opsin family. We demonstrate the following results. (1) The available vertebrate opsin sequences produce a definitive description of all five vertebrate opsin families. This is the first demonstration of sequence saturation prior to ET analysis, which we term saturated ET (SET). (2) The cone opsin classes have class-specific sites compared to the rod opsin class. These sites reside in the transmembrane region and tune the spectral sensitivity of each opsin class to its characteristic wavelength range. (3) The cytoplasmic loops, primarily responsible for signal transmission and inactivation, are essentially invariant in rod versus cone opsins. This indicates that the electrophysiological differences between rod and cone photoreceptors cannot be ascribed to differences in the protein interaction regions of the opsins. SET shows that chromophore binding and regeneration are the only aspects of opsin structure likely to have functionally significant differences between rods and cones, whereas excitatory and adaptational properties of the opsin families appear to be functionally invariant. PMID:15988624

  4. Rod opsin cDNA sequence from the sand goby (Pomatoschistus minutus) compared with those of other vertebrates.

    PubMed

    Archer, S N; Lythgoe, J N; Hall, L

    1992-04-22

    The absorbance spectra of rods from the sand goby were measured by using microspectrophotometry. Analysis of the averaged spectra shows that the rod visual pigment has a maximum absorbance (lambda max) at approximately 501 nm. A sand goby retinal cDNA library was constructed and then screened with a partial sand goby rod opsin clone obtained by the polymerase chain reaction (PCR). The screening of the library yielded a full length rod opsin clone. The cDNA sequence and deduced amino acid sequence of this clone are compared with those of other vertebrate rod opsins. PMID:1381834

  5. Opsins in Limulus eyes: characterization of three visible light-sensitive opsins unique to and co-expressed in median eye photoreceptors and a peropsin/RGR that is expressed in all eyes

    PubMed Central

    Battelle, Barbara-Anne; Kempler, Karen E.; Saraf, Spencer R.; Marten, Catherine E.; Dugger, Donald R.; Speiser, Daniel I.; Oakley, Todd H.

    2015-01-01

    The eyes of the horseshoe crab Limulus polyphemus have long been used for studies of basic mechanisms of vision, and the structure and physiology of Limulus photoreceptors have been examined in detail. Less is known about the opsins Limulus photoreceptors express. We previously characterized a UV opsin (LpUVOps1) that is expressed in all three types of Limulus eyes (lateral compound eyes, median ocelli and larval eyes) and three visible light-sensitive rhabdomeric opsins (LpOps1, -2 and -5) that are expressed in Limulus lateral compound and larval eyes. Physiological studies showed that visible light-sensitive photoreceptors are also present in median ocelli, but the visible light-sensitive opsins they express were unknown. In the current study we characterize three newly identified, visible light-sensitive rhabdomeric opsins (LpOps6, -7 and -8) that are expressed in median ocelli. We show that they are ocellar specific and that all three are co-expressed in photoreceptors distinct from those expressing LpUVOps1. Our current findings show that the pattern of opsin expression in Limulus eyes is much more complex than previously thought and extend our previous observations of opsin co-expression in visible light-sensitive Limulus photoreceptors. We also characterize a Limulus peropsin/RGR (LpPerOps1). We examine the phylogenetic relationship of LpPerOps1 with other peropsins and RGRs, demonstrate that LpPerOps1 transcripts are expressed in each of the three types of Limulus eyes and show that the encoded protein is expressed in membranes of cells closely associated with photoreceptors in each eye type. These finding suggest that peropsin was in the opsin repertoire of euchelicerates. PMID:25524988

  6. Genomic and gene regulatory signatures of cryptozoic adaptation: Loss of blue sensitive photoreceptors through expansion of long wavelength-opsin expression in the red flour beetle Tribolium castaneum

    PubMed Central

    Jackowska, Magdalena; Bao, Riyue; Liu, Zhenyi; McDonald, Elizabeth C; Cook, Tiffany A; Friedrich, Markus

    2007-01-01

    Background Recent genome sequence analysis in the red flour beetle Tribolium castaneum indicated that this highly crepuscular animal encodes only two single opsin paralogs: a UV-opsin and a long wavelength (LW)-opsin; however, these animals do not encode a blue (B)-opsin as most other insects. Here, we studied the spatial regulation of the Tribolium single LW- and UV-opsin gene paralogs in comparison to that of the five opsin paralogs in the retina of Drosophila melanogaster. Results In situ hybridization analysis reveals that the Tribolium retina, in contrast with other insect retinas, constitutes a homogenous field of ommatidia that have seven LW-opsin expressing photoreceptors and one UV-/LW-opsin co-expressing photoreceptor per eye unit. This pattern is consistent with the loss of photoreceptors sensitive to blue wavelengths. It also identifies Tribolium as the first example of a species in insects that co-expresses two different opsins across the entire retina in violation of the widely observed "one receptor rule" of sensory cells. Conclusion Broader studies of opsin evolution in darkling beetles and other coleopteran groups have the potential to pinpoint the permissive and adaptive forces that played a role in the evolution of vision in Tribolium castaneum. PMID:18154648

  7. De Novo Adult Transcriptomes of Two European Brittle Stars: Spotlight on Opsin-Based Photoreception.

    PubMed

    Delroisse, Jérôme; Mallefet, Jérôme; Flammang, Patrick

    2016-01-01

    Next generation sequencing (NGS) technology allows to obtain a deeper and more complete view of transcriptomes. For non-model or emerging model marine organisms, NGS technologies offer a great opportunity for rapid access to genetic information. In this study, paired-end Illumina HiSeqTM technology has been employed to analyse transcriptomes from the arm tissues of two European brittle star species, Amphiura filiformis and Ophiopsila aranea. About 48 million Illumina reads were generated and 136,387 total unigenes were predicted from A. filiformis arm tissues. For O. aranea arm tissues, about 47 million reads were generated and 123,324 total unigenes were obtained. Twenty-four percent of the total unigenes from A. filiformis show significant matches with sequences present in reference online databases, whereas, for O. aranea, this percentage amounts to 23%. In both species, around 50% of the predicted annotated unigenes were significantly similar to transcripts from the purple sea urchin, the closest species to date that has undergone complete genome sequencing and annotation. GO, COG and KEGG analyses were performed on predicted brittle star unigenes. We focused our analyses on the phototransduction actors involved in light perception. Firstly, two new echinoderm opsins were identified in O. aranea: one rhabdomeric opsin (homologous to vertebrate melanopsin) and one RGR opsin. The RGR-opsin is supposed to be involved in retinal regeneration while the r-opsin is suspected to play a role in visual-like behaviour. Secondly, potential phototransduction actors were identified in both transcriptomes using the fly (rhabdomeric) and mammal (ciliary) classical phototransduction pathways as references. Finally, the sensitivity of O.aranea to monochromatic light was investigated to complement data available for A. filiformis. The presence of microlens-like structures at the surface of dorsal arm plate of O. aranea could potentially explain phototactic behaviour differences between the two species. The results confirm (i) the ability of these brittle stars to perceive light using opsin-based photoreception, (ii) suggest the co-occurrence of both rhabdomeric and ciliary photoreceptors, and (iii) emphasise the complexity of light perception in this echinoderm class. PMID:27119739

  8. De Novo Adult Transcriptomes of Two European Brittle Stars: Spotlight on Opsin-Based Photoreception

    PubMed Central

    Mallefet, Jérôme; Flammang, Patrick

    2016-01-01

    Next generation sequencing (NGS) technology allows to obtain a deeper and more complete view of transcriptomes. For non-model or emerging model marine organisms, NGS technologies offer a great opportunity for rapid access to genetic information. In this study, paired-end Illumina HiSeqTM technology has been employed to analyse transcriptomes from the arm tissues of two European brittle star species, Amphiura filiformis and Ophiopsila aranea. About 48 million Illumina reads were generated and 136,387 total unigenes were predicted from A. filiformis arm tissues. For O. aranea arm tissues, about 47 million reads were generated and 123,324 total unigenes were obtained. Twenty-four percent of the total unigenes from A. filiformis show significant matches with sequences present in reference online databases, whereas, for O. aranea, this percentage amounts to 23%. In both species, around 50% of the predicted annotated unigenes were significantly similar to transcripts from the purple sea urchin, the closest species to date that has undergone complete genome sequencing and annotation. GO, COG and KEGG analyses were performed on predicted brittle star unigenes. We focused our analyses on the phototransduction actors involved in light perception. Firstly, two new echinoderm opsins were identified in O. aranea: one rhabdomeric opsin (homologous to vertebrate melanopsin) and one RGR opsin. The RGR-opsin is supposed to be involved in retinal regeneration while the r-opsin is suspected to play a role in visual-like behaviour. Secondly, potential phototransduction actors were identified in both transcriptomes using the fly (rhabdomeric) and mammal (ciliary) classical phototransduction pathways as references. Finally, the sensitivity of O.aranea to monochromatic light was investigated to complement data available for A. filiformis. The presence of microlens-like structures at the surface of dorsal arm plate of O. aranea could potentially explain phototactic behaviour differences between the two species. The results confirm (i) the ability of these brittle stars to perceive light using opsin-based photoreception, (ii) suggest the co-occurrence of both rhabdomeric and ciliary photoreceptors, and (iii) emphasise the complexity of light perception in this echinoderm class. PMID:27119739

  9. Adaptation of pineal expressed teleost exo-rod opsin to non-image forming photoreception through enhanced Meta II decay.

    PubMed

    Tarttelin, Emma E; Fransen, Maikel P; Edwards, Patricia C; Hankins, Mark W; Schertler, Gebhard F X; Vogel, Reiner; Lucas, Robert J; Bellingham, James

    2011-11-01

    Photoreception by vertebrates enables both image-forming vision and non-image-forming responses such as circadian photoentrainment. Over the recent years, distinct non-rod non-cone photopigments have been found to support circadian photoreception in diverse species. By allowing specialization to this sensory task a selective advantage is implied, but the nature of that specialization remains elusive. We have used the presence of distinct rod opsin genes specialized to either image-forming (retinal rod opsin) or non-image-forming (pineal exo-rod opsin) photoreception in ray-finned fish (Actinopterygii) to gain a unique insight into this problem. A comparison of biochemical features for these paralogous opsins in two model teleosts, Fugu pufferfish (Takifugu rubripes) and zebrafish (Danio rerio), reveals striking differences. While spectral sensitivity is largely unaltered by specialization to the pineal environment, in other aspects exo-rod opsins exhibit a behavior that is quite distinct from the cardinal features of the rod opsin family. While they display a similar thermal stability, they show a greater than tenfold reduction in the lifetime of the signaling active Meta II photoproduct. We show that these features reflect structural changes in retinal association domains of helices 3 and 5 but, interestingly, not at either of the two residues known to define these characteristics in cone opsins. Our findings suggest that the requirements of non-image-forming photoreception have lead exo-rod opsin to adopt a characteristic that seemingly favors efficient bleach recovery but not at the expense of absolute sensitivity. PMID:21416149

  10. In vivo incorporation of (2-/sup 3/H)-myo-inositol into frog opsin

    SciTech Connect

    Fliesler, S.J.; Anderson, R.E.

    1986-04-29

    The in vivo incorporation of (2-/sup 3/H)-myo-inositol into frog retinal rod outer segment membranes was examined. About 25% of the recovered radioactivity was found to be protein-associated. Following acid hydrolysis of this material and extraction with hexane, all the radioactivity remained in the aqueous phase, indicating that the label was not in fatty acids. Following ion exchange column chromatography of the hydrolysate, the major radioactive compound comigrated on TLC with an internal standard of (U-/sup 14/C)-myo-inositol. SDS polyacrylamide gel electrophoresis of unextracted membranes indicated that the majority of the label was associated with opsin. These results indicate that (2-/sup 3/H)-myo-inositol was incorporated in vivo into opsin, presumably with retention of its chemical identity.

  11. Imaging neural spiking in brain tissue using FRET-opsin protein voltage sensors

    PubMed Central

    Gong, Yiyang; Wagner, Mark J.; Li, Jin Zhong; Schnitzer, Mark J.

    2014-01-01

    Genetically encoded fluorescence voltage sensors offer the possibility of directly visualizing neural spiking dynamics in cells targeted by their genetic class or connectivity. Sensors of this class have generally suffered performance-limiting tradeoffs between modest brightness, sluggish kinetics, and limited signaling dynamic range in response to action potentials. Here we describe sensors that use fluorescence resonance energy transfer (FRET) to combine the rapid kinetics and substantial voltage-dependence of rhodopsin family voltage-sensing domains with the brightness of genetically engineered protein fluorophores. These FRET-opsin sensors significantly improve upon the spike detection fidelity offered by the genetically encoded voltage sensor, Arclight, while offering faster kinetics and higher brightness. Using FRET-opsin sensors we imaged neural spiking and sub-threshold membrane voltage dynamics in cultured neurons and in pyramidal cells within neocortical tissue slices. In live mice, rates and optical waveforms of cerebellar Purkinje neurons’ dendritic voltage transients matched expectations for these cells’ dendritic spikes. PMID:24755708

  12. A Phe-rich region in short-wavelength sensitive opsins is responsible for their aggregation in the absence of 11-cis-retinal.

    PubMed

    Zhang, Tao; Fu, Yingbin

    2013-08-01

    Human blue and mouse S-opsin are prone to aggregation in the absence of 11-cis-retinal, which underlie the rapid cone degeneration in human patients and animal models of Leber congenital amaurosis (LCA). By in silico analysis and domain swapping experiments, we show that a Phe-rich region in short-wavelength sensitive (SWS) opsins, but not in medium/long-wavelength sensitive opsins, is responsible for SWS opsin aggregation. Mutagenesis studies suggest that Phe residues in this region are critical in mediating protein aggregation. Fusing the Phe-rich region of SWS opsins to GFP causes the latter to aggregate. Our findings suggest that new therapeutics can be designed to disrupt the Phe-rich region in preventing cone degeneration due to S-opsin aggregation in LCA. PMID:23792161

  13. A Phe-rich region in short-wavelength sensitive opsins is responsible for their aggregation in the absence of 11-cis-retinal

    PubMed Central

    Zhang, Tao; Fu, Yingbin

    2013-01-01

    Human blue and mouse S-opsin are prone to aggregation in the absence of 11-cis-retinal, which underlie the rapid cone degeneration in human patients and animal models of Leber congenital amaurosis (LCA). By in silico analysis and domain swapping experiments, we show that a Phe-rich region in short-wavelength sensitive (SWS) opsins, but not in medium/long-wavelength sensitive opsins, is responsible for SWS opsin aggregation. Mutagenesis studies suggest that Phe residues in this region are critical in mediating protein aggregation. Fusing the Phe-rich region of SWS opsins to GFP causes the latter to aggregate. Our findings suggest that new therapeutics can be designed to disrupt the Phe-rich region in preventing cone degeneration due to S-opsin aggregation in LCA. PMID:23792161

  14. Ancestral duplications and highly dynamic opsin gene evolution in percomorph fishes

    PubMed Central

    Cortesi, Fabio; Musilová, Zuzana; Stieb, Sara M.; Hart, Nathan S.; Siebeck, Ulrike E.; Malmstrøm, Martin; Tørresen, Ole K.; Jentoft, Sissel; Cheney, Karen L.; Marshall, N. Justin; Carleton, Karen L.; Salzburger, Walter

    2015-01-01

    Single-gene and whole-genome duplications are important evolutionary mechanisms that contribute to biological diversification by launching new genetic raw material. For example, the evolution of animal vision is tightly linked to the expansion of the opsin gene family encoding light-absorbing visual pigments. In teleost fishes, the most species-rich vertebrate group, opsins are particularly diverse and key to the successful colonization of habitats ranging from the bioluminescence-biased but basically dark deep sea to clear mountain streams. In this study, we report a previously unnoticed duplication of the violet-blue short wavelength-sensitive 2 (SWS2) opsin, which coincides with the radiation of highly diverse percomorph fishes, permitting us to reinterpret the evolution of this gene family. The inspection of close to 100 fish genomes revealed that, triggered by frequent gene conversion between duplicates, the evolutionary history of SWS2 is rather complex and difficult to predict. Coincidentally, we also report potential cases of gene resurrection in vertebrate opsins, whereby pseudogenized genes were found to convert with their functional paralogs. We then identify multiple novel amino acid substitutions that are likely to have contributed to the adaptive differentiation between SWS2 copies. Finally, using the dusky dottyback Pseudochromis fuscus, we show that the newly discovered SWS2A duplicates can contribute to visual adaptation in two ways: by gaining sensitivities to different wavelengths of light and by being differentially expressed between ontogenetic stages. Thus, our study highlights the importance of comparative approaches in gaining a comprehensive view of the dynamics underlying gene family evolution and ultimately, animal diversification. PMID:25548152

  15. Phenotypic plasticity in opsin expression in a butterfly compound eye complements sex role reversal

    PubMed Central

    2012-01-01

    Background Animals often display phenotypic plasticity in morphologies and behaviors that result in distinct adaptations to fluctuating seasonal environments. The butterfly Bicyclus anynana has two seasonal forms, wet and dry, that vary in wing ornament brightness and in the identity of the sex that performs the most courting and choosing. Rearing temperature is the cue for producing these alternative seasonal forms. We hypothesized that, barring any developmental constraints, vision should be enhanced in the choosy individuals but diminished in the non-choosy individuals due to physiological costs. As a proxy of visual performance we measured eye size, facet lens size, and sensitivity to light, e.g., the expression levels of all opsins, in males and females of both seasonal forms. Results We found that B. anynana eyes displayed significant sexual dimorphism and phenotypic plasticity for both morphology and opsin expression levels, but not all results conformed to our prediction. Males had larger eyes than females across rearing temperatures, and increases in temperature produced larger eyes in both sexes, mostly via increases in facet number. Ommatidia were larger in the choosy dry season (DS) males and transcript levels for all three opsins were significantly lower in the less choosy DS females. Conclusions Opsin level plasticity in females, and ommatidia size plasticity in males supported our visual plasticity hypothesis but males appear to maintain high visual function across both seasons. We discuss our results in the context of distinct sexual and natural selection pressures that may be facing each sex in the wild in each season. PMID:23194112

  16. Ancestral duplications and highly dynamic opsin gene evolution in percomorph fishes.

    PubMed

    Cortesi, Fabio; Musilová, Zuzana; Stieb, Sara M; Hart, Nathan S; Siebeck, Ulrike E; Malmstrøm, Martin; Tørresen, Ole K; Jentoft, Sissel; Cheney, Karen L; Marshall, N Justin; Carleton, Karen L; Salzburger, Walter

    2015-02-01

    Single-gene and whole-genome duplications are important evolutionary mechanisms that contribute to biological diversification by launching new genetic raw material. For example, the evolution of animal vision is tightly linked to the expansion of the opsin gene family encoding light-absorbing visual pigments. In teleost fishes, the most species-rich vertebrate group, opsins are particularly diverse and key to the successful colonization of habitats ranging from the bioluminescence-biased but basically dark deep sea to clear mountain streams. In this study, we report a previously unnoticed duplication of the violet-blue short wavelength-sensitive 2 (SWS2) opsin, which coincides with the radiation of highly diverse percomorph fishes, permitting us to reinterpret the evolution of this gene family. The inspection of close to 100 fish genomes revealed that, triggered by frequent gene conversion between duplicates, the evolutionary history of SWS2 is rather complex and difficult to predict. Coincidentally, we also report potential cases of gene resurrection in vertebrate opsins, whereby pseudogenized genes were found to convert with their functional paralogs. We then identify multiple novel amino acid substitutions that are likely to have contributed to the adaptive differentiation between SWS2 copies. Finally, using the dusky dottyback Pseudochromis fuscus, we show that the newly discovered SWS2A duplicates can contribute to visual adaptation in two ways: by gaining sensitivities to different wavelengths of light and by being differentially expressed between ontogenetic stages. Thus, our study highlights the importance of comparative approaches in gaining a comprehensive view of the dynamics underlying gene family evolution and ultimately, animal diversification. PMID:25548152

  17. Enhancer/promoter activities of the long/middle wavelength-sensitive opsins of vertebrates mediated by thyroid hormone receptor β2 and COUP-TFII.

    PubMed

    Iwagawa, Toshiro; Tanaka, Yo; Iida, Atsumi; Itoh, Toshio; Watanabe, Sumiko

    2013-01-01

    Cone photopigments (opsins) are crucial elements of, and the first detection module in, color vision. Individual opsins have different wavelength sensitivity patterns, and the temporal and spatial expression patterns of opsins are unique and stringently regulated. Long and middle wavelength-sensitive (L/M) opsins are of the same phylogenetic type. Although the roles of thyroid hormone/TRß2 and COUP-TFs in the transcriptional regulation of L/M opsins have been explored, the detailed mechanisms, including the target sequence in the enhancer of L/M opsins, have not been revealed. We aimed to reveal molecular mechanisms of L/M opsins in vertebrates. Using several human red opsin enhancer/promoter-luciferase reporter constructs, we found that TRß2 increased luciferase activities through the 5'-UTR and intron 3-4 region, whereas the presence of T3 affected only the intron 3-4 region-dependent luciferase activity. Furthermore, COUP-TFII suppressed intron 3-4 region-dependent luciferase activities. However, luciferase expression driven by the mouse M opsin intron 3-4 region was only slightly increased by TRß2, and rather enhanced by COUP-TFII. To determine whether these differential responses reflect differences between primates and rodents, we examined the enhancer/promoter region of the red opsin of the common marmoset. Interestingly, while TRß2 increased 5'-UTR- or intron 3-4 region-driven luciferase expression, as observed for the human red opsin, expression of the latter luciferase was not suppressed by COUP-TFII. In fact, immunostaining of common marmoset retinal sections revealed expression of COUP-TFII and red opsin in the cone cells. PMID:24058409

  18. Atomistic design of microbial opsin-based blue-shifted optogenetics tools

    PubMed Central

    Kato, Hideaki E.; Kamiya, Motoshi; Sugo, Seiya; Ito, Jumpei; Taniguchi, Reiya; Orito, Ayaka; Hirata, Kunio; Inutsuka, Ayumu; Yamanaka, Akihiro; Maturana, Andrés D.; Ishitani, Ryuichiro; Sudo, Yuki; Hayashi, Shigehiko; Nureki, Osamu

    2015-01-01

    Microbial opsins with a bound chromophore function as photosensitive ion transporters and have been employed in optogenetics for the optical control of neuronal activity. Molecular engineering has been utilized to create colour variants for the functional augmentation of optogenetics tools, but was limited by the complexity of the protein–chromophore interactions. Here we report the development of blue-shifted colour variants by rational design at atomic resolution, achieved through accurate hybrid molecular simulations, electrophysiology and X-ray crystallography. The molecular simulation models and the crystal structure reveal the precisely designed conformational changes of the chromophore induced by combinatory mutations that shrink its π-conjugated system which, together with electrostatic tuning, produce large blue shifts of the absorption spectra by maximally 100 nm, while maintaining photosensitive ion transport activities. The design principle we elaborate is applicable to other microbial opsins, and clarifies the underlying molecular mechanism of the blue-shifted action spectra of microbial opsins recently isolated from natural sources. PMID:25975962

  19. Atomistic design of microbial opsin-based blue-shifted optogenetics tools.

    PubMed

    Kato, Hideaki E; Kamiya, Motoshi; Sugo, Seiya; Ito, Jumpei; Taniguchi, Reiya; Orito, Ayaka; Hirata, Kunio; Inutsuka, Ayumu; Yamanaka, Akihiro; Maturana, Andrés D; Ishitani, Ryuichiro; Sudo, Yuki; Hayashi, Shigehiko; Nureki, Osamu

    2015-01-01

    Microbial opsins with a bound chromophore function as photosensitive ion transporters and have been employed in optogenetics for the optical control of neuronal activity. Molecular engineering has been utilized to create colour variants for the functional augmentation of optogenetics tools, but was limited by the complexity of the protein-chromophore interactions. Here we report the development of blue-shifted colour variants by rational design at atomic resolution, achieved through accurate hybrid molecular simulations, electrophysiology and X-ray crystallography. The molecular simulation models and the crystal structure reveal the precisely designed conformational changes of the chromophore induced by combinatory mutations that shrink its π-conjugated system which, together with electrostatic tuning, produce large blue shifts of the absorption spectra by maximally 100 nm, while maintaining photosensitive ion transport activities. The design principle we elaborate is applicable to other microbial opsins, and clarifies the underlying molecular mechanism of the blue-shifted action spectra of microbial opsins recently isolated from natural sources. PMID:25975962

  20. Luminopsins integrate opto- and chemogenetics by using physical and biological light sources for opsin activation

    PubMed Central

    Berglund, Ken; Clissold, Kara; Li, Haofang E.; Wen, Lei; Park, Sung Young; Gleixner, Jan; Klein, Marguerita E.; Lu, Dongye; Barter, Joseph W.; Rossi, Mark A.; Augustine, George J.; Yin, Henry H.; Hochgeschwender, Ute

    2016-01-01

    Luminopsins are fusion proteins of luciferase and opsin that allow interrogation of neuronal circuits at different temporal and spatial resolutions by choosing either extrinsic physical or intrinsic biological light for its activation. Building on previous development of fusions of wild-type Gaussia luciferase with channelrhodopsin, here we expanded the utility of luminopsins by fusing bright Gaussia luciferase variants with either channelrhodopsin to excite neurons (luminescent opsin, LMO) or a proton pump to inhibit neurons (inhibitory LMO, iLMO). These improved LMOs could reliably activate or silence neurons in vitro and in vivo. Expression of the improved LMO in hippocampal circuits not only enabled mapping of synaptic activation of CA1 neurons with fine spatiotemporal resolution but also could drive rhythmic circuit excitation over a large spatiotemporal scale. Furthermore, virus-mediated expression of either LMO or iLMO in the substantia nigra in vivo produced not only the expected bidirectional control of single unit activity but also opposing effects on circling behavior in response to systemic injection of a luciferase substrate. Thus, although preserving the ability to be activated by external light sources, LMOs expand the use of optogenetics by making the same opsins accessible to noninvasive, chemogenetic control, thereby allowing the same probe to manipulate neuronal activity over a range of spatial and temporal scales. PMID:26733686

  1. Atomistic design of microbial opsin-based blue-shifted optogenetics tools

    NASA Astrophysics Data System (ADS)

    Kato, Hideaki E.; Kamiya, Motoshi; Sugo, Seiya; Ito, Jumpei; Taniguchi, Reiya; Orito, Ayaka; Hirata, Kunio; Inutsuka, Ayumu; Yamanaka, Akihiro; Maturana, Andrés D.; Ishitani, Ryuichiro; Sudo, Yuki; Hayashi, Shigehiko; Nureki, Osamu

    2015-05-01

    Microbial opsins with a bound chromophore function as photosensitive ion transporters and have been employed in optogenetics for the optical control of neuronal activity. Molecular engineering has been utilized to create colour variants for the functional augmentation of optogenetics tools, but was limited by the complexity of the protein-chromophore interactions. Here we report the development of blue-shifted colour variants by rational design at atomic resolution, achieved through accurate hybrid molecular simulations, electrophysiology and X-ray crystallography. The molecular simulation models and the crystal structure reveal the precisely designed conformational changes of the chromophore induced by combinatory mutations that shrink its π-conjugated system which, together with electrostatic tuning, produce large blue shifts of the absorption spectra by maximally 100 nm, while maintaining photosensitive ion transport activities. The design principle we elaborate is applicable to other microbial opsins, and clarifies the underlying molecular mechanism of the blue-shifted action spectra of microbial opsins recently isolated from natural sources.

  2. Luminopsins integrate opto- and chemogenetics by using physical and biological light sources for opsin activation.

    PubMed

    Berglund, Ken; Clissold, Kara; Li, Haofang E; Wen, Lei; Park, Sung Young; Gleixner, Jan; Klein, Marguerita E; Lu, Dongye; Barter, Joseph W; Rossi, Mark A; Augustine, George J; Yin, Henry H; Hochgeschwender, Ute

    2016-01-19

    Luminopsins are fusion proteins of luciferase and opsin that allow interrogation of neuronal circuits at different temporal and spatial resolutions by choosing either extrinsic physical or intrinsic biological light for its activation. Building on previous development of fusions of wild-type Gaussia luciferase with channelrhodopsin, here we expanded the utility of luminopsins by fusing bright Gaussia luciferase variants with either channelrhodopsin to excite neurons (luminescent opsin, LMO) or a proton pump to inhibit neurons (inhibitory LMO, iLMO). These improved LMOs could reliably activate or silence neurons in vitro and in vivo. Expression of the improved LMO in hippocampal circuits not only enabled mapping of synaptic activation of CA1 neurons with fine spatiotemporal resolution but also could drive rhythmic circuit excitation over a large spatiotemporal scale. Furthermore, virus-mediated expression of either LMO or iLMO in the substantia nigra in vivo produced not only the expected bidirectional control of single unit activity but also opposing effects on circling behavior in response to systemic injection of a luciferase substrate. Thus, although preserving the ability to be activated by external light sources, LMOs expand the use of optogenetics by making the same opsins accessible to noninvasive, chemogenetic control, thereby allowing the same probe to manipulate neuronal activity over a range of spatial and temporal scales. PMID:26733686

  3. IFT20 is required for opsin trafficking and photoreceptor outer segment development.

    PubMed

    Keady, Brian T; Le, Yun Zheng; Pazour, Gregory J

    2011-04-01

    The light-detecting outer segments of vertebrate photoreceptors are cilia. Like other cilia, all materials needed for assembly and maintenance are synthesized in the cell body and transported into the cilium. The highly elaborated nature of the outer segment and its high rate of turnover necessitate unusually high levels of transport into the cilium. In this work, we examine the role of the IFT20 subunit of the intraflagellar transport (IFT) particle in photoreceptor cells. IFT20 was deleted in developing cones by a cone-specific Cre and in mature rods and cones by a tamoxifen-activatable Cre. Loss of IFT20 during cone development leads to opsin accumulation in the inner segment even when the connecting cilium and outer segment are still intact. With time this causes cone cell degeneration. Similarly, deletion of IFT20 in mature rods causes rapid accumulation of rhodopsin in the cell body, where it is concentrated at the Golgi complex. We further show that IFT20, acting both as part of the IFT particle and independent of the particle, binds to rhodopsin and RG-opsin. Since IFT20 dynamically moves between the Golgi complex and the connecting cilium, the current work suggests that rhodopsin and opsins are cargo for IFT transport. PMID:21307337

  4. Point mutations in bovine opsin can be classified in four groups with respect to their effect on the biosynthetic pathway of opsin.

    PubMed Central

    DeCaluwé, G L; DeGrip, W J

    1996-01-01

    Expression in vitro with the recombinant baculovirus expression system showed correct biosynthesis and post-translational processing of "wild-type' bovine opsin with regard to translocation, glycosylation, palmitoylation and targeting. However, several of these processes were severely affected by point mutations. From the overall results of 16 mutants reported here, four groups were distinguished. One group significantly affected neither biosynthesis nor folding of opsin (D83N, P291A, A299C-V300A-P303G). A second group produced a truncated protein (R69H, Y301F), suggesting that these positions are essential for a correct translational process. A third group affected membrane translocation as well as glycosylation, which can be interpreted as interference with the function of a transfer signal. Substitutions at positions Glu-113, Glu-122, Glu-134, Arg-135 and Lys-248 belong to this category. A fourth group induced structural changes in the protein that led to heterogeneous distribution in the plasma membrane (E113Q/D, W265F, Y268S). Taking any functional consequences of these mutations into consideration, it seems that point mutations can have mosaic effects and therefore should be examined at several levels (folding, targeting, functional parameters). PMID:9003366

  5. Evolutionary renovation of L/M opsin polymorphism confers a fruit discrimination advantage to ateline New World monkeys.

    PubMed

    Matsumoto, Yoshifumi; Hiramatsu, Chihiro; Matsushita, Yuka; Ozawa, Norihiro; Ashino, Ryuichi; Nakata, Makiko; Kasagi, Satoshi; Di Fiore, Anthony; Schaffner, Colleen M; Aureli, Filippo; Melin, Amanda D; Kawamura, Shoji

    2014-04-01

    New World monkeys exhibit prominent colour vision variation due to allelic polymorphism of the long-to-middle wavelength (L/M) opsin gene. The known spectral variation of L/M opsins in primates is broadly determined by amino acid composition at three sites: 180, 277 and 285 (the 'three-sites' rule). However, two L/M opsin alleles found in the black-handed spider monkeys (Ateles geoffroyi) are known exceptions, presumably due to novel mutations. The spectral separation of the two L/M photopigments is 1.5 times greater than expected based on the 'three-sites' rule. Yet the consequence of this for the visual ecology of the species is unknown, as is the evolutionary mechanism by which spectral shift was achieved. In this study, we first examine L/M opsins of two other Atelinae species, the long-haired spider monkeys (A. belzebuth) and the common woolly monkeys (Lagothrix lagotricha). By a series of site-directed mutagenesis, we show that a mutation Y213D (tyrosine to aspartic acid at site 213) in the ancestral opsin of the two alleles enabled N294K, which occurred in one allele of the ateline ancestor and increased the spectral separation between the two alleles. Second, by modelling the chromaticity of dietary fruits and background leaves in a natural habitat of spider monkeys, we demonstrate that chromatic discrimination of fruit from leaves is significantly enhanced by these mutations. This evolutionary renovation of L/M opsin polymorphism in atelines illustrates a previously unappreciated dynamism of opsin genes in shaping primate colour vision. PMID:24612406

  6. Evolutionary renovation of L/M opsin polymorphism confers a fruit discrimination advantage to ateline New World monkeys

    PubMed Central

    Matsumoto, Yoshifumi; Hiramatsu, Chihiro; Matsushita, Yuka; Ozawa, Norihiro; Ashino, Ryuichi; Nakata, Makiko; Kasagi, Satoshi; Di Fiore, Anthony; Schaffner, Colleen M; Aureli, Filippo; Melin, Amanda D; Kawamura, Shoji

    2014-01-01

    New World monkeys exhibit prominent colour vision variation due to allelic polymorphism of the long-to-middle wavelength (L/M) opsin gene. The known spectral variation of L/M opsins in primates is broadly determined by amino acid composition at three sites: 180, 277 and 285 (the ‘three-sites’ rule). However, two L/M opsin alleles found in the black-handed spider monkeys (Ateles geoffroyi) are known exceptions, presumably due to novel mutations. The spectral separation of the two L/M photopigments is 1.5 times greater than expected based on the ‘three-sites’ rule. Yet the consequence of this for the visual ecology of the species is unknown, as is the evolutionary mechanism by which spectral shift was achieved. In this study, we first examine L/M opsins of two other Atelinae species, the long-haired spider monkeys (A. belzebuth) and the common woolly monkeys (Lagothrix lagotricha). By a series of site-directed mutagenesis, we show that a mutation Y213D (tyrosine to aspartic acid at site 213) in the ancestral opsin of the two alleles enabled N294K, which occurred in one allele of the ateline ancestor and increased the spectral separation between the two alleles. Second, by modelling the chromaticity of dietary fruits and background leaves in a natural habitat of spider monkeys, we demonstrate that chromatic discrimination of fruit from leaves is significantly enhanced by these mutations. This evolutionary renovation of L/M opsin polymorphism in atelines illustrates a previously unappreciated dynamism of opsin genes in shaping primate colour vision. PMID:24612406

  7. Genetic deletion of S-opsin prevents rapid cone degeneration in a mouse model of Leber congenital amaurosis

    PubMed Central

    Zhang, Tao; Enemchukwu, Nduka O.; Jones, Alex; Wang, Shixian; Dennis, Emily; Watt, Carl B.; Pugh, Edward N.; Fu, Yingbin

    2015-01-01

    Mutations in RPE65 or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal synthesis and cause Leber congenital amaurosis (LCA), a severe hereditary blindness occurring in early childhood. The pathology is attributed to a combination of 11-cis-retinal deficiency and photoreceptor degeneration. The mistrafficking of cone membrane-associated proteins including cone opsins (M- and S-opsins), cone transducin (Gαt2), G-protein-coupled receptor kinase 1 (GRK1) and guanylate cyclase 1 (GC1) has been suggested to play a role in cone degeneration. However, their precise role in cone degeneration is unclear. Here we investigated the role of S-opsin (Opn1sw) in cone degeneration in Lrat−/−, a murine model for LCA, by genetic ablation of S-opsin. We show that deletion of just one allele of S-opsin from Lrat−/− mice is sufficient to prevent the rapid cone degeneration for at least 1 month. Deletion of both alleles of S-opsin prevents cone degeneration for an extended period (at least 12 months). This genetic prevention is accompanied by a reduction of endoplasmic reticulum (ER) stress in Lrat−/− photoreceptors. Despite cone survival in Opn1sw−/−Lrat−/− mice, cone membrane-associated proteins (e.g. Gαt2, GRK1 and GC1) continue to have trafficking problems. Our results suggest that cone opsins are the ‘culprit’ linking 11-cis-retinal deficiency to cone degeneration in LCA. This result has important implications for the current gene therapy strategy that emphasizes the need for a combinatorial therapy to both improve vision and slow photoreceptor degeneration. PMID:25416279

  8. Genetic deletion of S-opsin prevents rapid cone degeneration in a mouse model of Leber congenital amaurosis.

    PubMed

    Zhang, Tao; Enemchukwu, Nduka O; Jones, Alex; Wang, Shixian; Dennis, Emily; Watt, Carl B; Pugh, Edward N; Fu, Yingbin

    2015-03-15

    Mutations in RPE65 or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal synthesis and cause Leber congenital amaurosis (LCA), a severe hereditary blindness occurring in early childhood. The pathology is attributed to a combination of 11-cis-retinal deficiency and photoreceptor degeneration. The mistrafficking of cone membrane-associated proteins including cone opsins (M- and S-opsins), cone transducin (Gαt2), G-protein-coupled receptor kinase 1 (GRK1) and guanylate cyclase 1 (GC1) has been suggested to play a role in cone degeneration. However, their precise role in cone degeneration is unclear. Here we investigated the role of S-opsin (Opn1sw) in cone degeneration in Lrat(-) (/-), a murine model for LCA, by genetic ablation of S-opsin. We show that deletion of just one allele of S-opsin from Lrat(-) (/-) mice is sufficient to prevent the rapid cone degeneration for at least 1 month. Deletion of both alleles of S-opsin prevents cone degeneration for an extended period (at least 12 months). This genetic prevention is accompanied by a reduction of endoplasmic reticulum (ER) stress in Lrat(-) (/-) photoreceptors. Despite cone survival in Opn1sw(-/-)Lrat(-) (/-) mice, cone membrane-associated proteins (e.g. Gαt2, GRK1 and GC1) continue to have trafficking problems. Our results suggest that cone opsins are the 'culprit' linking 11-cis-retinal deficiency to cone degeneration in LCA. This result has important implications for the current gene therapy strategy that emphasizes the need for a combinatorial therapy to both improve vision and slow photoreceptor degeneration. PMID:25416279

  9. P23H opsin knock-in mice reveal a novel step in retinal rod disc morphogenesis.

    PubMed

    Sakami, Sanae; Kolesnikov, Alexander V; Kefalov, Vladimir J; Palczewski, Krzysztof

    2014-04-01

    Retinal rod photoreceptor cells have double membrane discs located in their outer segments (ROS) that are continuously formed proximally from connecting cilia (CC) and phagocytized distally by the retinal pigmented epithelium. The major component of these rod discs, the light-sensitive visual pigment rhodopsin (Rho), consists of an opsin protein linked to 11-cis-retinal. The P23H mutation of rod opsin (P23H opsin) is the most common cause of human blinding autosomal dominant retinitis pigmentosa (adRP). A mouse model of adRP with this mutation (Rho(P23H/+)) shows low levels of P23H opsin protein, partial misalignment of discs and progressive retinal degeneration. However, the impact of mutant P23H opsin on the formation of abnormal discs is unclear and it is still unknown whether this mutant pigment can mediate phototransduction. Using transretinal ERG recordings, we demonstrate that P23H mutant Rho can trigger phototransduction but Rho(P23H/P23H) rods are ∼17 000-fold less sensitive to light than Rho(+/+) rods and produce abnormally fast photo-responses. By analyzing homozygous Rho(P23H/P23H) knock-in mice, we show that P23H opsin is transported to ciliary protrusions where it forms sagittally elongated discs. Transmission electron microscopy of postnatal day (PND) 14 Rho(P23H/+) mouse retina revealed disordered sagittally oriented discs before the onset of retinal degeneration. Surprisingly, we also observed smaller, immature sagittally oriented discs in PND14 Rho(+/)(-) and Rho(+/+) mice that were not seen in older animals. These findings provide fundamental insights into the pathogenesis of the P23H mutant opsin and reveal a novel early sagittally aligned disc formation step in normal ROS disc expansion. PMID:24214395

  10. 11-cis-retinal reduces constitutive opsin phosphorylation and improves quantum catch in retinoid-deficient mouse rod photoreceptors.

    PubMed

    Ablonczy, Zsolt; Crouch, Rosalie K; Goletz, Patrice W; Redmond, T Michael; Knapp, Daniel R; Ma, Jian-Xing; Rohrer, Barbel

    2002-10-25

    Rpe65(-/-) mice produce minimal amounts of 11-cis-retinal, the ligand necessary for the formation of photosensitive visual pigments. Therefore, the apoprotein opsin in these animals has not been exposed to its normal ligand. The Rpe65(-/-) mice contain less than 0.1% of wild type levels of rhodopsin. Mass spectrometric analysis of opsin from Rpe65(-/-) mice revealed unusually high levels of phosphorylation in dark-adapted mice but no other structural alterations. Single flash and flicker electroretinograms (ERGs) from 1-month-old animals showed trace rod function but no cone response. B-wave kinetics of the single-flash ERG are comparable with those of dark-adapted wild type mice containing a full compliment of rhodopsin. Application (intraperitoneal injection) of 11-cis-retinal to Rpe65(-/-) mice increased the rod ERG signal, increased levels of rhodopsin, and decreased opsin phosphorylation. Therefore, exogenous 11-cis-retinal improves photoreceptor function by regenerating rhodopsin and removes constitutive opsin phosphorylation. Our results indicate that opsin, which has not been exposed to 11-cis-retinal, does not generate the activity generally associated with the bleached apoprotein. PMID:12176991

  11. Travelling-wave Green tensor and near-field Rayleigh-wave sensitivity

    NASA Astrophysics Data System (ADS)

    Liu, Kui; Zhou, Ying

    2016-04-01

    Travelling-wave Green tensor has been widely used in calculations of synthetic seismograms and finite-frequency sensitivities of surface waves. The classic travelling-wave decomposition is based on a far-field approximation and may not be valid when applied to construct sensitivity kernels in regions close to the receiver. In this paper, we calculate synthetic seismograms and finite-frequency sensitivity kernels of Rayleigh waves based on travelling-wave representation of Green tensor that fully accounts for near-field effects. We show that far-field approximation is adequate for synthetic seismograms when the source-receiver epicentral distance is greater than the dominant wavelength. Errors in Rayleigh-wave sensitivity kernels introduced by far-field approximation are in general negligible for single-station measurements except for in a small region around the station, and the errors are more significant in sensitivity kernels for interstation measurements. In addition, interstation measurements are strongly sensitive to structures outside the region between the two stations, even for two stations along the same great circle path from the seismic source.

  12. Metal Mesh Fabrication and Testing for Infrared Astronomy and ISO Science Programs; ISO GO Data Analysis and LWS Instrument Team Activities

    NASA Technical Reports Server (NTRS)

    Smith, Howard A.; Oliversen, Ronald J. (Technical Monitor)

    2001-01-01

    This research program addresses astrophysics research with the Infrared Space Observatory's Long Wavelength Spectrometer (ISO-LWS), including efforts to supply ISO-LWS with superior metal mesh filters. This grant has, over the years, enabled Dr. Smith in his role as a Co-Investigator on the satellite, the PI (Principal Investigator) on the Extragalactic Science Team, and a member of the Calibration and performance working groups. The emphasis of the budget in this proposal is in support of Dr. Smith's Infrared Space Observatory research. This program began (under a different grant number) while Dr. Smith was at the Smithsonian's National Air and Space Museum, and was transferred to SAO with a change in number. While Dr. Smith was a visiting Discipline Scientist at NASA HQ the program was in abeyance, but it has resumed in full since his return to SAO. The Infrared Space Observatory mission was launched in November, 1996, and since then has successfully completed its planned lifetime mission. Data are currently being calibrated to the 2% level.

  13. Opsins in onychophora (velvet worms) suggest a single origin and subsequent diversification of visual pigments in arthropods.

    PubMed

    Hering, Lars; Henze, Miriam J; Kohler, Martin; Kelber, Almut; Bleidorn, Christoph; Leschke, Maren; Nickel, Birgit; Meyer, Matthias; Kircher, Martin; Sunnucks, Paul; Mayer, Georg

    2012-11-01

    Multiple visual pigments, prerequisites for color vision, are found in arthropods, but the evolutionary origin of their diversity remains obscure. In this study, we explore the opsin genes in five distantly related species of Onychophora, using deep transcriptome sequencing and screening approaches. Surprisingly, our data reveal the presence of only one opsin gene (onychopsin) in each onychophoran species, and our behavioral experiments indicate a maximum sensitivity of onychopsin to blue-green light. In our phylogenetic analyses, the onychopsins represent the sister group to the monophyletic clade of visual r-opsins of arthropods. These results concur with phylogenomic support for the sister-group status of the Onychophora and Arthropoda and provide evidence for monochromatic vision in velvet worms and in the last common ancestor of Onychophora and Arthropoda. We conclude that the diversification of visual pigments and color vision evolved in arthropods, along with the evolution of compound eyes-one of the most sophisticated visual systems known. PMID:22683812

  14. Nonvisual Opsins and the Regulation of Peripheral Clocks by Light and Hormones.

    PubMed

    Poletini, Maristela O; Ramos, Bruno C; Moraes, Maria Nathalia; Castrucci, Ana Maria L

    2015-01-01

    The molecular clock machinery is conserved throughout evolution. However, how environmental cues are perceived has evolved in such a way that peripheral clocks in mammals require a variety of signals, including hormones. On the other hand, in nonmammalian cells able to directly detect light, light seems to play a major role in the synchronization of the clock. The interaction between perception of circadian light by nonvisual opsins and hormones will be discussed under the perspective of clock synchronization at the molecular level. PMID:26174318

  15. Co-Expression of VAL- and TMT-Opsins Uncovers Ancient Photosensory Interneurons and Motorneurons in the Vertebrate Brain

    PubMed Central

    Fischer, Ruth M.; Fontinha, Bruno M.; Kirchmaier, Stephan; Steger, Julia; Bloch, Susanne; Inoue, Daigo; Panda, Satchidananda; Rumpel, Simon; Tessmar-Raible, Kristin

    2013-01-01

    The functional principle of the vertebrate brain is often paralleled to a computer: information collected by dedicated devices is processed and integrated by interneuron circuits and leads to output. However, inter- and motorneurons present in today's vertebrate brains are thought to derive from neurons that combined sensory, integration, and motor function. Consistently, sensory inter­motorneurons have been found in the simple nerve nets of cnidarians, animals at the base of the evolutionary lineage. We show that light-sensory motorneurons and light-sensory interneurons are also present in the brains of vertebrates, challenging the paradigm that information processing and output circuitry in the central brain is shielded from direct environmental influences. We investigated two groups of nonvisual photopigments, VAL- and TMT-Opsins, in zebrafish and medaka fish; two teleost species from distinct habitats separated by over 300 million years of evolution. TMT-Opsin subclasses are specifically expressed not only in hypothalamic and thalamic deep brain photoreceptors, but also in interneurons and motorneurons with no known photoreceptive function, such as the typeXIV interneurons of the fish optic tectum. We further show that TMT-Opsins and Encephalopsin render neuronal cells light-sensitive. TMT-Opsins preferentially respond to blue light relative to rhodopsin, with subclass-specific response kinetics. We discovered that tmt-opsins co-express with val-opsins, known green light receptors, in distinct inter- and motorneurons. Finally, we show by electrophysiological recordings on isolated adult tectal slices that interneurons in the position of typeXIV neurons respond to light. Our work supports “sensory-inter-motorneurons” as ancient units for brain evolution. It also reveals that vertebrate inter- and motorneurons are endowed with an evolutionarily ancient, complex light-sensory ability that could be used to detect changes in ambient light spectra, possibly providing the endogenous equivalent to an optogenetic machinery. PMID:23776409

  16. NinaB is essential for Drosophila vision but induces retinal degeneration in opsin-deficient photoreceptors.

    PubMed

    Voolstra, Olaf; Oberhauser, Vitus; Sumser, Emerich; Meyer, Nina E; Maguire, Michael E; Huber, Armin; von Lintig, Johannes

    2010-01-15

    In animals, visual pigments are essential for photoreceptor function and survival. These G-protein-coupled receptors consist of a protein moiety (opsin) and a covalently bound 11-cis-retinylidene chromophore. The chromophore is derived from dietary carotenoids by oxidative cleavage and trans-to-cis isomerization of double bonds. In vertebrates, the necessary chemical transformations are catalyzed by two distinct but structurally related enzymes, the carotenoid oxygenase beta-carotenoid-15,15'-monooxygenase and the retinoid isomerase RPE65 (retinal pigment epithelium protein of 65 kDa). Recently, we provided biochemical evidence that these reactions in insects are catalyzed by a single enzyme family member named NinaB. Here we show that in the fly pathway, carotenoids are mandatory precursors of the chromophore. After chromophore formation, the retinoid-binding protein Pinta acts downstream of NinaB and is required to supply photoreceptors with chromophore. Like ninaE encoding the opsin, ninaB expression is eye-dependent and is activated as a downstream target of the eyeless/pax6 and sine oculis master control genes for eye development. The requirement for coordinated synthesis of chromophore and opsin is evidenced by analysis of ninaE mutants. Retinal degeneration in opsin-deficient photoreceptors is caused by the chromophore and can be prevented by restricting its supply as seen in an opsin and chromophore-deficient double mutant. Thus, our study identifies NinaB as a key component for visual pigment production and provides evidence that chromophore in opsin-deficient photoreceptors can elicit retinal degeneration. PMID:19889630

  17. Membrane assembly in retinal photoreceptors. II. Immunocytochemical analysis of freeze-fractured rod photoreceptor membranes using anti-opsin antibodies.

    PubMed

    Defoe, D M; Besharse, J C

    1985-04-01

    We have used a cytochemical technique for labeling freeze-fractured tissues (Pinto da Silva, P., C. Parkison, and N. Dwyer (1981) Proc. Natl. Acad. Sci. U.S.A. 78: 343-347) to examine the distribution of immunoreactive opsin in rod photoreceptor membranes. Aldehyde-fixed retinas of African clawed frogs (Xenopus laevis) embedded in a cross-linked protein matrix were frozen and fractured at -196 degrees C, then thawed and labeled with biotinylated sheep anti-cow opsin IgG followed by avidin-ferritin. In thin sections of plastic-embedded retinas, rod outer segment (ROS) disc membranes exposed by fracturing bound specific antibody intensely and relatively uniformly. However, they differed from membranes of the inner segment as well as those of erythrocytes in that protoplasmic face leaflets did not assume an interrupted bilayer appearance and disc exoplasmic face leaflets were apparently lost during thawing. The disposition of opsin immunoreactivity in the cell membrane was highly asymmetric. Although ROS plasma membranes from which discs are elaborated labeled heavily with anti-opsin after cleavage, fractures passing along inner segment plasma membranes bound very little antibody. In cross-fractures exposing inner segment cytoplasm, we found specific labeling of Golgi complex elements, as well as both perimitochondrial and periciliary vesicles. The latter are presumed to be the vehicle shuttling newly synthesized membrane to the ROS for disc assembly. These results suggest that opsin-containing membrane is sorted out within the cell, being transported from synthetic sites to the immediate periciliary zone where localized insertion into the cell membrane takes place. Furthermore, the close correspondence of the present immunocytochemical analysis with the distribution of opsin deduced from prior quantitative freeze-fracture analysis (Besharse, J. C., and K. H. Pfenninger (1980) J. Cell Biol. 87: 451-463) offers the possibility that fracture-label may be generally useful for study of patterned membrane topography in neuronal cells. PMID:3156972

  18. Blue-light-receptive cryptochrome is expressed in a sponge eye lacking neurons and opsin

    PubMed Central

    Rivera, Ajna S.; Ozturk, Nuri; Fahey, Bryony; Plachetzki, David C.; Degnan, Bernard M.; Sancar, Aziz; Oakley, Todd H.

    2012-01-01

    SUMMARY Many larval sponges possess pigment ring eyes that apparently mediate phototactic swimming. Yet sponges are not known to possess nervous systems or opsin genes, so the unknown molecular components of sponge phototaxis must differ fundamentally from those in other animals, inspiring questions about how this sensory system functions. Here we present molecular and biochemical data on cryptochrome, a candidate gene for functional involvement in sponge pigment ring eyes. We report that Amphimedon queenslandica, a demosponge, possesses two cryptochrome/photolyase genes, Aq-Cry1 and Aq-Cry2. The mRNA of one gene (Aq-Cry2) is expressed in situ at the pigment ring eye. Additionally, we report that Aq-Cry2 lacks photolyase activity and contains a flavin-based co-factor that is responsive to wavelengths of light that also mediate larval photic behavior. These results suggest that Aq-Cry2 may act in the aneural, opsin-less phototaxic behavior of a sponge. PMID:22442365

  19. Organization of the G Protein-coupled Receptors Rhodopsin and Opsin in Native Membranes*

    PubMed Central

    Filipek, Sławomir; Saperstein, David A.; Palczewski, Krzysztof; Engel, Andreas

    2006-01-01

    G protein-coupled receptors (GPCRs), which constitute the largest and structurally best conserved family of signaling molecules, are involved in virtually all physiological processes. Crystal structures are available only for the detergent-solubilized light receptor rhodopsin. In addition, this receptor is the only GPCR for which the presumed higher order oligomeric state in native membranes has been demonstrated (Fotiadis, D., Liang, Y., Filipek, S., Saperstein, D. A., Engel, A., and Palczewski, K. (2003) Nature 421, 127–128). Here, we have determined by atomic force microscopy the organization of rhodopsin in native membranes obtained from wild-type mouse photoreceptors and opsin isolated from photoreceptors of Rpe65−/− mutant mice, which do not produce the chromophore 11-cis-retinal. The higher order organization of rhodopsin was present irrespective of the support on which the membranes were adsorbed for imaging. Rhodopsin and opsin form structural dimers that are organized in paracrystal-line arrays. The intradimeric contact is likely to involve helices IV and V, whereas contacts mainly between helices I and II and the cytoplasmic loop connecting helices V and VI facilitate the formation of rhodopsin dimer rows. Contacts between rows are on the extracellular side and involve helix I. This is the first semi-empirical model of a higher order structure of a GPCR in native membranes, and it has profound implications for the understanding of how this receptor interacts with partner proteins. PMID:12663652

  20. The evolution of irradiance detection: melanopsin and the non-visual opsins

    PubMed Central

    Peirson, Stuart N.; Halford, Stephanie; Foster, Russell G.

    2009-01-01

    Circadian rhythms are endogenous 24 h cycles that persist in the absence of external time cues. These rhythms provide an internal representation of day length and optimize physiology and behaviour to the varying demands of the solar cycle. These clocks require daily adjustment to local time and the primary time cue (zeitgeber) used by most vertebrates is the daily change in the amount of environmental light (irradiance) at dawn and dusk, a process termed photoentrainment. Attempts to understand the photoreceptor mechanisms mediating non-image-forming responses to light, such as photoentrainment, have resulted in the discovery of a remarkable array of different photoreceptors and photopigment families, all of which appear to use a basic opsin/vitamin A-based photopigment biochemistry. In non-mammalian vertebrates, specialized photoreceptors are located within the pineal complex, deep brain and dermal melanophores. There is also strong evidence in fish and amphibians for the direct photic regulation of circadian clocks in multiple tissues. By contrast, mammals possess only ocular photoreceptors. However, in addition to the image-forming rods and cones of the retina, there exists a third photoreceptor system based on a subset of melanopsin-expressing photosensitive retinal ganglion cells (pRGCs). In this review, we discuss the range of vertebrate photoreceptors and their opsin photopigments, describe the melanopsin/pRGC system in some detail and then finally consider the molecular evolution and sensory ecology of these non-image-forming photoreceptor systems. PMID:19720649

  1. A human opsin-related gene that encodes a retinaldehyde-binding protein.

    PubMed

    Shen, D; Jiang, M; Hao, W; Tao, L; Salazar, M; Fong, H K

    1994-11-01

    The ligand-binding property of a cytoplasmic membrane-bound protein from bovine retinal pigment epithelium (RPE) has been demonstrated. The putative RPE-retinal G protein coupled receptor (RGR) covalently binds both all-trans- and 11-cis-retinal after reduction by sodium borohydride. The 32-kDa receptor binds all-trans-retinal preferentially, rather than the 11-cis isomer. The amino acid sequence of the opsin-related protein in humans is 86% identical to that of bovine RGR, and a lysine residue, analogous to the retinaldehyde attachment site of rhodopsin, is conserved in the seventh transmembrane domain of RGR in both species. The human gene that encodes the novel retinaldehyde receptor spans 14.8 kb and is split into seven exons. The structure of the gene is distinct from that of the visual pigment genes. These findings support the notion that the rgr gene represents the earliest independent branch of the vertebrate opsin gene family. A second form of human RGR in retina is predicted by alternative splicing of its precursor mRNA. This RGR variant results from the alternative use of an internal acceptor splice site in the second intron of the human gene, and it contains an insertion of four amino acids in the connecting loop between the second and thrid transmembrane domains. Since RGR binds all-trans-retinal preferentially, one of its functions may be to catalyze isomerization of the chromophore by a retinochrome-like mechanism. PMID:7947717

  2. Jellyfish vision starts with cAMP signaling mediated by opsin-G(s) cascade.

    PubMed

    Koyanagi, Mitsumasa; Takano, Kosuke; Tsukamoto, Hisao; Ohtsu, Kohzoh; Tokunaga, Fumio; Terakita, Akihisa

    2008-10-01

    Light sensing starts with phototransduction in photoreceptor cells. The phototransduction cascade has diverged in different species, such as those mediated by transducin in vertebrate rods and cones, by G(q)-type G protein in insect and molluscan rhabdomeric-type visual cells and vertebrate photosensitive retinal ganglion cells, and by G(o)-type G protein in scallop ciliary-type visual cells. Here, we investigated the phototransduction cascade of a prebilaterian box jellyfish, the most basal animal having eyes containing lens and ciliary-type visual cells similar to vertebrate eyes, to examine the similarity at the molecular level and to obtain an implication of the origin of the vertebrate phototransduction cascade. We showed that the opsin-based pigment functions as a green-sensitive visual pigment and triggers the G(s)-type G protein-mediated phototransduction cascade in the ciliary-type visual cells of the box jellyfish lens eyes. We also demonstrated the light-dependent cAMP increase in the jellyfish visual cells and HEK293S cells expressing the jellyfish opsin. The first identified prebilaterian cascade was distinct from known phototransduction cascades but exhibited significant partial similarity with those in vertebrate and molluscan ciliary-type visual cells, because all involved cyclic nucleotide signaling. These similarities imply a monophyletic origin of ciliary phototransduction cascades distributed from prebilaterian to vertebrate. PMID:18832159

  3. Blue-light-receptive cryptochrome is expressed in a sponge eye lacking neurons and opsin.

    PubMed

    Rivera, Ajna S; Ozturk, Nuri; Fahey, Bryony; Plachetzki, David C; Degnan, Bernard M; Sancar, Aziz; Oakley, Todd H

    2012-04-15

    Many larval sponges possess pigment ring eyes that apparently mediate phototactic swimming. Yet sponges are not known to possess nervous systems or opsin genes, so the unknown molecular components of sponge phototaxis must differ fundamentally from those in other animals, inspiring questions about how this sensory system functions. Here we present molecular and biochemical data on cryptochrome, a candidate gene for functional involvement in sponge pigment ring eyes. We report that Amphimedon queenslandica, a demosponge, possesses two cryptochrome/photolyase genes, Aq-Cry1 and Aq-Cry2. The mRNA of one gene (Aq-Cry2) is expressed in situ at the pigment ring eye. Additionally, we report that Aq-Cry2 lacks photolyase activity and contains a flavin-based co-factor that is responsive to wavelengths of light that also mediate larval photic behavior. These results suggest that Aq-Cry2 may act in the aneural, opsin-less phototaxic behavior of a sponge. PMID:22442365

  4. Strategies for identification of mutations causing hereditary retinal diseases in dogs: evaluation of opsin as a candidate gene.

    PubMed

    Ray, K; Wang, W; Czarnecki, J; Zhang, Q; Acland, G M; Aguirre, G D

    1999-01-01

    Progressive retinal atrophy (PRA), like retinitis pigmentosa (RP) in man, represents a clinical classification grouping together a variety of hereditary diseases of the visual cells which have broadly similar clinical characteristics. At least six distinct autosomal recessive and one X-linked retinal disease locus have been identified. As one of the strategies to look for the gene defect causing the different forms of PRA, we are examining first the most promising candidate genes. These include those coding for photoreceptor-specific structural proteins and enzymes of the phototransduction pathway, especially those reported to cause RP. Preeminent among these candidates is the gene for rod opsin, in which multiple causative mutations have been identified in both dominant and recessive forms of RP. In addition, mutations in this gene are also causally associated with congenital stationary night blindness (CSNB) in man. We have used two strategies to examine the rod opsin gene for association with inherited retinal disease in dogs: (1) linkage to determine cosegregation of the disease locus with an intragenic polymorphic marker in the opsin gene in those breeds where suitable informative pedigrees were available; and (2) scanning the coding sequence of the gene in cases where only a limited number of affected or obligate heterozygous samples were available for a breed. We conclude that mutations in the rod opsin gene are not associated with PRA or CSNB in the 11 different dog breeds tested. PMID:9987920

  5. Reproducible and sustained regulation of Gαs signalling using a metazoan opsin as an optogenetic tool.

    PubMed

    Bailes, Helena J; Zhuang, Ling-Yu; Lucas, Robert J

    2012-01-01

    Originally developed to regulate neuronal excitability, optogenetics is increasingly also used to control other cellular processes with unprecedented spatiotemporal resolution. Optogenetic modulation of all major G-protein signalling pathways (Gq, Gi and Gs) has been achieved using variants of mammalian rod opsin. We show here that the light response driven by such rod opsin-based tools dissipates under repeated exposure, consistent with the known bleaching characteristics of this photopigment. We continue to show that replacing rod opsin with a bleach resistant opsin from Carybdea rastonii, the box jellyfish, (JellyOp) overcomes this limitation. Visible light induced high amplitude, reversible, and reproducible increases in cAMP in mammalian cells expressing JellyOp. While single flashes produced a brief cAMP spike, repeated stimulation could sustain elevated levels for 10s of minutes. JellyOp was more photosensitive than currently available optogenetic tools, responding to white light at irradiances ≥1 µW/cm(2). We conclude that JellyOp is a promising new tool for mimicking the activity of Gs-coupled G protein coupled receptors with fine spatiotemporal resolution. PMID:22292038

  6. Genomic organization, evolution, and expression of photoprotein and opsin genes in Mnemiopsis leidyi: a new view of ctenophore photocytes

    PubMed Central

    2012-01-01

    Background Calcium-activated photoproteins are luciferase variants found in photocyte cells of bioluminescent jellyfish (Phylum Cnidaria) and comb jellies (Phylum Ctenophora). The complete genomic sequence from the ctenophore Mnemiopsis leidyi, a representative of the earliest branch of animals that emit light, provided an opportunity to examine the genome of an organism that uses this class of luciferase for bioluminescence and to look for genes involved in light reception. To determine when photoprotein genes first arose, we examined the genomic sequence from other early-branching taxa. We combined our genomic survey with gene trees, developmental expression patterns, and functional protein assays of photoproteins and opsins to provide a comprehensive view of light production and light reception in Mnemiopsis. Results The Mnemiopsis genome has 10 full-length photoprotein genes situated within two genomic clusters with high sequence conservation that are maintained due to strong purifying selection and concerted evolution. Photoprotein-like genes were also identified in the genomes of the non-luminescent sponge Amphimedon queenslandica and the non-luminescent cnidarian Nematostella vectensis, and phylogenomic analysis demonstrated that photoprotein genes arose at the base of all animals. Photoprotein gene expression in Mnemiopsis embryos begins during gastrulation in migrating precursors to photocytes and persists throughout development in the canals where photocytes reside. We identified three putative opsin genes in the Mnemiopsis genome and show that they do not group with well-known bilaterian opsin subfamilies. Interestingly, photoprotein transcripts are co-expressed with two of the putative opsins in developing photocytes. Opsin expression is also seen in the apical sensory organ. We present evidence that one opsin functions as a photopigment in vitro, absorbing light at wavelengths that overlap with peak photoprotein light emission, raising the hypothesis that light production and light reception may be functionally connected in ctenophore photocytes. We also present genomic evidence of a complete ciliary phototransduction cascade in Mnemiopsis. Conclusions This study elucidates the genomic organization, evolutionary history, and developmental expression of photoprotein and opsin genes in the ctenophore Mnemiopsis leidyi, introduces a novel dual role for ctenophore photocytes in both bioluminescence and phototransduction, and raises the possibility that light production and light reception are linked in this early-branching non-bilaterian animal. PMID:23259493

  7. Parallel reduction in expression, but no loss of functional constraint, in two opsin paralogs within cave populations of Gammarus minus (Crustacea: Amphipoda)

    PubMed Central

    2013-01-01

    Background Gammarus minus, a freshwater amphipod living in the cave and surface streams in the eastern USA, is a premier candidate for studying the evolution of troglomorphic traits such as pigmentation loss, elongated appendages, and reduced eyes. In G. minus, multiple pairs of genetically related, physically proximate cave and surface populations exist which exhibit a high degree of intraspecific morphological divergence. The morphology, ecology, and genetic structure of these sister populations are well characterized, yet the genetic basis of their morphological divergence remains unknown. Results We used degenerate PCR primers designed to amplify opsin genes within the subphylum Crustacea and discovered two distinct opsin paralogs (average inter-paralog protein divergence ≈ 20%) in the genome of three independently derived pairs of G. minus cave and surface populations. Both opsin paralogs were found to be related to other crustacean middle wavelength sensitive opsins. Low levels of nucleotide sequence variation (< 1% within populations) were detected in both opsin genes, regardless of habitat, and dN/dS ratios did not indicate a relaxation of functional constraint in the cave populations with reduced or absent eyes. Maximum likelihood analyses using codon-based models also did not detect a relaxation of functional constraint in the cave lineages. We quantified expression level of both opsin genes and found that the expression of both paralogs was significantly reduced in all three cave populations relative to their sister surface populations. Conclusions The concordantly lowered expression level of both opsin genes in cave populations of G. minus compared to sister surface populations, combined with evidence for persistent purifying selection in the cave populations, is consistent with an unspecified pleiotropic function of opsin proteins. Our results indicate that phototransduction proteins such as opsins may have retained their function in cave-adapted organisms because they may play a pleiotropic role in other important processes that are unrelated to vision. PMID:23617561

  8. Mislocalized Opsin and cAMP Signaling: A Mechanism for Sprouting by Rod Cells in Retinal Degeneration

    PubMed Central

    Wang, Jianfeng; Zhang, Nan; Beuve, Annie; Townes-Anderson, Ellen

    2012-01-01

    Purpose. In human retinal degeneration, rod photoreceptors reactively sprout neurites. The mechanism is unknown in part because of the paucity of animal models displaying this feature of human pathology. We tested the role of cAMP and opsin in sprouting by tiger salamander rod cells, photoreceptors that can produce reactive growth. Methods. In vitro systems of isolated photoreceptor cells and intact neural retina were used. cAMP signaling was manipulated with nucleotide analogues, enzyme stimulators, agonists for adenosine and dopamine receptors, and the opsin agonist, ?-ionone. Levels of cAMP were determined by radioimmunoassay, and protein levels by Western blot and quantitative immunocytochemistry. Neuritic growth was assayed by image analysis and conventional and confocal microscopy. Results. cAMP analogues and stimulation of adenylyl cyclase (AC) directly or through G-proteincoupled receptors resulted in significant increases in neuritic growth of isolated rod, but not cone, cells. The signaling pathway included protein kinase A (PKA) and phosphorylation of the transcription factor cAMP response element-binding protein (pCREB). Opsin, a G-linked receptor, is present throughout the plasmalemma of isolated cells; its activation also induced sprouting. In neural retina, rod sprouting was significantly increased by ?-ionone with concomitant increases in cAMP, pCREB, and synaptic proteins. Notably, opsin stimulated sprouting only when mislocalized to the plasmalemma of the rod cell body. Conclusions. cAMP causes neuritic sprouting in rod, but not cone, cells through the AC-PKA-CREB pathway known to be associated with synaptic plasticity. We propose that in retinal disease, mislocalized rod opsin gains access to cAMP signaling, which leads to neuritic sprouting. PMID:22899763

  9. A Non-Mammalian Type Opsin 5 Functions Dually in the Photoreceptive and Non-Photoreceptive Organs of Birds

    PubMed Central

    Ohuchi, Hideyo; Yamashita, Takahiro; Tomonari, Sayuri; Fujita-Yanagibayashi, Sari; Sakai, Kazumi; Noji, Sumihare; Shichida, Yoshinori

    2012-01-01

    A mammalian type opsin 5 (neuropsin) is a recently identified ultraviolet (UV)-sensitive pigment of the retina and other photosensitive organs in birds. Two other opsin 5-related molecules have been found in the genomes of non-mammalian vertebrates. However, their functions have not been examined as yet. Here, we identify the molecular properties of a second avian opsin 5, cOpn5L2 (chicken opsin 5-like 2), and its localization in the post-hatch chicken. Spectrophotometric analysis and radionucleotide-binding assay have revealed that cOpn5L2 is a UV-sensitive bistable pigment that couples with the Gi subtype of guanine nucleotide-binding protein (G protein). As a bistable pigment, it also shows the direct binding ability to agonist all-trans-retinal to activate G protein. The absorption maxima of UV-light-absorbing and visible light-absorbing forms were 350 and 521 nm, respectively. Expression analysis showed relatively high expression of cOpn5L2 mRNA in the adrenal gland, which is not photoreceptive but an endocrine organ, while lower expression was found in the brain and retina. At the protein level, cOpn5L2 immunoreactive cells were present in the chromaffin cells of the adrenal gland. In the brain, cOpn5L2 immunoreactive cells were found in the paraventricular and supraoptic nuclei of the anterior hypothalamus, known for photoreceptive deep brain areas. In the retina, cOpn5L2 protein was localized to subsets of cells in the ganglion cell layer and the inner nuclear layer. These results suggest that the non-mammalian type opsin 5 (Opn5L2) functions as a second UV sensor in the photoreceptive organs, while it might function as chemosensor using its direct binding ability to agonist all-trans-retinal in non-photoreceptive organs such as the adrenal gland of birds. PMID:22348098

  10. Opsin activation of transduction in the rods of dark-reared Rpe65 knockout mice.

    PubMed

    Fan, Jie; Woodruff, Michael L; Cilluffo, Marianne C; Crouch, Rosalie K; Fain, Gordon L

    2005-10-01

    Rpe65 knockout mice (Rpe65-/-) are unable to synthesize the visual pigment chromophore 11-cis retinal; however, if these animals are reared in complete darkness, the rod photoreceptors accumulate a small amount of 9-cis retinal and its corresponding visual pigment isorhodopsin. Suction-electrode recording of single rods from dark-reared Rpe65-/- mice showed that the rods were about 400 times less sensitive than wild-type control rods and that the maximum responses were much smaller in amplitude. Spectral sensitivity measurements indicated that Rpe65-/- rod responses were generated by isorhodopsin rather than rhodopsin. Sensitivity and pigment concentration were compared in the same mice by measuring light responses from rods of one eye and pigment concentration from the retina of the other eye. Retinas had 11-35% of the normal pigment level, but the rods were of the order of 20-30 times less sensitive than could be accounted for by the loss in quantum catch. This extra desensitization must be caused by opsin-dependent activation of the visual cascade, which leads to a state equivalent to light adaptation in the dark-adapted rod. By comparing the sensitivity of dark-reared Rpe65-/- rods to that produced in normal rods by background light, we estimate that Rpe65-/- opsin is of the order of 2.5x10(-5) as efficient in activating transduction as photoactivated rhodopsin (Rh*) in WT mice. Dark-reared Rpe65-/- rods are less desensitized than rods from cyclic light-reared Rpe65-/- mice, have about 50% more photocurrent and degenerate at a slower rate. Retinas sectioned after 9 months in darkness show a larger number of photoreceptor nuclei in dark-reared animals than in cyclic light-reared animals, though both have fewer nuclei than in cyclic light-reared wild-type retinas. Both also have shorter outer segments and a lower free-Ca2+ concentration. These experiments provide the first quantitative measurement of opsin activation in physiologically responding mammalian rods. PMID:15994181

  11. A blind circadian clock in cavefish reveals that opsins mediate peripheral clock photoreception.

    PubMed

    Cavallari, Nicola; Frigato, Elena; Vallone, Daniela; Fröhlich, Nadine; Lopez-Olmeda, Jose Fernando; Foà, Augusto; Berti, Roberto; Sánchez-Vázquez, Francisco Javier; Bertolucci, Cristiano; Foulkes, Nicholas S

    2011-09-01

    The circadian clock is synchronized with the day-night cycle primarily by light. Fish represent fascinating models for deciphering the light input pathway to the vertebrate clock since fish cell clocks are regulated by direct light exposure. Here we have performed a comparative, functional analysis of the circadian clock involving the zebrafish that is normally exposed to the day-night cycle and a cavefish species that has evolved in perpetual darkness. Our results reveal that the cavefish retains a food-entrainable clock that oscillates with an infradian period. Importantly, however, this clock is not regulated by light. This comparative study pinpoints the two extra-retinal photoreceptors Melanopsin (Opn4m2) and TMT-opsin as essential upstream elements of the peripheral clock light input pathway. PMID:21909239

  12. Optical control of neuronal excitation and inhibition using a single opsin protein, ChR2

    PubMed Central

    Liske, Holly; Qian, Xiang; Anikeeva, Polina; Deisseroth, Karl; Delp, Scott

    2013-01-01

    The effect of electrical stimulation on neuronal membrane potential is frequency dependent. Low frequency electrical stimulation can evoke action potentials, whereas high frequency stimulation can inhibit action potential transmission. Optical stimulation of channelrhodopsin-2 (ChR2) expressed in neuronal membranes can also excite action potentials. However, it is unknown whether optical stimulation of ChR2-expressing neurons produces a transition from excitation to inhibition with increasing light pulse frequencies. Here we report optical inhibition of motor neuron and muscle activity in vivo in the cooled sciatic nerves of Thy1-ChR2-EYFP mice. We also demonstrate all-optical single-wavelength control of neuronal excitation and inhibition without co-expression of inhibitory and excitatory opsins. This all-optical system is free from stimulation-induced electrical artifacts and thus provides a new approach to investigate mechanisms of high frequency inhibition in neuronal circuits in vivo and in vitro. PMID:24173561

  13. Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins

    PubMed Central

    Mattis, Joanna; Tye, Kay M; Ferenczi, Emily A; Ramakrishnan, Charu; O’Shea, Daniel J; Prakash, Rohit; Gunaydin, Lisa A; Hyun, Minsuk; Fenno, Lief E; Gradinaru, Viviana; Yizhar, Ofer; Deisseroth, Karl

    2014-01-01

    Diverse optogenetic tools have allowed versatile control over neural activity. Many depolarizing and hyperpolarizing tools have now been developed in multiple laboratories and tested across different preparations, presenting opportunities but also making it difficult to draw direct comparisons. This challenge has been compounded by the dependence of performance on parameters such as vector, promoter, expression time, illumination, cell type and many other variables. As a result, it has become increasingly complicated for end users to select the optimal reagents for their experimental needs. For a rapidly growing field, critical figures of merit should be formalized both to establish a framework for further development and so that end users can readily understand how these standardized parameters translate into performance. Here we systematically compared microbial opsins under matched experimental conditions to extract essential principles and identify key parameters for the conduct, design and interpretation of experiments involving optogenetic techniques. PMID:22179551

  14. The Expression of Three Opsin Genes from the Compound Eye of Helicoverpa armigera (Lepidoptera: Noctuidae) Is Regulated by a Circadian Clock, Light Conditions and Nutritional Status

    PubMed Central

    Yan, Shuo; Zhu, Jialin; Zhu, Weilong; Zhang, Xinfang; Li, Zhen; Liu, Xiaoxia; Zhang, Qingwen

    2014-01-01

    Visual genes may become inactive in species that inhabit poor light environments, and the function and regulation of opsin components in nocturnal moths are interesting topics. In this study, we cloned the ultraviolet (UV), blue (BL) and long-wavelength-sensitive (LW) opsin genes from the compound eye of the cotton bollworm and then measured their mRNA levels using quantitative real-time PCR. The mRNA levels fluctuated over a daily cycle, which might be an adaptation of a nocturnal lifestyle, and were dependent on a circadian clock. Cycling of opsin mRNA levels was disturbed by constant light or constant darkness, and the UV opsin gene was up-regulated after light exposure. Furthermore, the opsin genes tended to be down-regulated upon starvation. Thus, this study illustrates that opsin gene expression is determined by multiple endogenous and exogenous factors and is adapted to the need for nocturnal vision, suggesting that color vision may play an important role in the sensory ecology of nocturnal moths. PMID:25353953

  15. The effect of white light on normal and malignant murine melanocytes: A link between opsins, clock genes, and melanogenesis.

    PubMed

    de Assis, L V M; Moraes, M N; da Silveira Cruz-Machado, S; Castrucci, A M L

    2016-06-01

    The skin possesses a photosensitive system comprised of opsins whose function is not fully understood, and clock genes which exert an important regulatory role in skin biology. Here, we evaluated the presence of opsins in normal (Melan-a cells) and malignant (B16-F10 cells) murine melanocytes. Both cell lines express Opn2, Opn4 - for the first time reported in these cell types - as well as S-opsin. OPN4 protein was found in a small area capping the cell nuclei of B16-F10 cells kept in constant dark (DD); twenty-four hours after the white light pulse (WLP), OPN4 was found in the cell membrane. Despite the fact that B16-F10 cells expressed less Opn2 and Opn4 than Melan-a cells, our data indicate that the malignant melanocytes exhibited increased photoresponsiveness. The clock gene machinery is also severely downregulated in B16-F10 cells as compared to Melan-a cells. Per1, Per2, and Bmal1 expression increased in B16-F10 cells in response to WLP. Although no response in clock gene expression to WLP was observed in Melan-a cells, gene correlational data suggest a minor effect of WLP. In contrast to opsins and clock genes, melanogenesis is significantly upregulated in malignant melanocytes in comparison to Melan-a cells. Tyrosinase expression increased after WLP only in B16-F10 cells; however no increase in melanin content after WLP was seen in either cell line. Our findings may prove useful in the treatment and the development of new pharmacological approaches of depigmentation diseases and skin cancer. PMID:26947915

  16. Evolutionary dynamics of Rh2 opsins in birds demonstrate an episode of accelerated evolution in the New World warblers (Setophaga).

    PubMed

    Bloch, Natasha I; Price, Trevor D; Chang, Belinda S W

    2015-05-01

    Low rates of sequence evolution associated with purifying selection can be interrupted by episodic changes in selective regimes. Visual pigments are a unique system in which we can investigate the functional consequences of genetic changes, therefore connecting genotype to phenotype in the context of natural and sexual selection pressures. We study the RH2 and RH1 visual pigments (opsins) across 22 bird species belonging to two ecologically convergent clades, the New World warblers (Parulidae) and Old World warblers (Phylloscopidae) and evaluate rates of evolution in these clades along with data from 21 additional species. We demonstrate generally slow evolution of these opsins: both Rh1 and Rh2 are highly conserved across Old World and New World warblers. However, Rh2 underwent a burst of evolution within the New World genus Setophaga, where it accumulated substitutions at 6 amino acid sites across the species we studied. Evolutionary analyses revealed a significant increase in dN /dS in Setophaga, implying relatively strong selective pressures to overcome long-standing purifying selection. We studied the effects of each substitution on spectral tuning and found they do not cause large spectral shifts. Thus, substitutions may reflect other aspects of opsin function, such as those affecting photosensitivity and/or dark-light adaptation. Although it is unclear what these alterations mean for colour perception, we suggest that rapid evolution is linked to sexual selection, given the exceptional plumage colour diversification in Setophaga. PMID:25827331

  17. Sea urchin tube feet are photosensory organs that express a rhabdomeric-like opsin and PAX6.

    PubMed

    Lesser, Michael P; Carleton, Karen L; Böttger, Stefanie A; Barry, Thomas M; Walker, Charles W

    2011-11-22

    All echinoderms have unique hydraulic structures called tube feet, known for their roles in light sensitivity, respiration, chemoreception and locomotion. In the green sea urchin, the most distal portion of these tube feet contain five ossicles arranged as a light collector with its concave surface facing towards the ambient light. These ossicles are perforated and lined with pigment cells that express a PAX6 protein that is universally involved in the development of eyes and sensory organs in other bilaterians. Polymerase chain reaction (PCR)-based sequencing and real time quantitative PCR (qPCR) also demonstrate the presence and differential expression of a rhabdomeric-like opsin within these tube feet. Morphologically, nerves that could serve to transmit information to the test innervate the tube feet, and the differential expression of opsin transcripts in the tube feet is inversely, and significantly, related to the amount of light that tube feet are exposed to depending on their location on the test. The expression of these genes, the differential expression of opsin based on light exposure and the unique morphological features at the distal portion of the tube foot strongly support the hypothesis that in addition to previously identified functional roles of tube feet they are also photosensory organs that detect and respond to changes in the underwater light field. PMID:21450733

  18. Evolutionary dynamics of Rh2 opsins in birds demonstrate an episode of accelerated evolution in the New World warblers (Setophaga)

    PubMed Central

    Price, Trevor D.

    2015-01-01

    Low rates of sequence evolution associated with purifying selection can be interrupted by episodic changes in selective regimes. Visual pigments are a unique system in which we can investigate the functional consequences of genetic changes, therefore connecting genotype to phenotype in the context of natural and sexual selection pressures. We study the RH2 and RH1 visual pigments (opsins) across 22 bird species belonging to two ecologically convergent clades, the New World warblers (Parulidae) and Old World warblers (Phylloscopidae), and evaluate rates of evolution in these clades along with data from 21 additional species. We demonstrate generally slow evolution of these opsins: both Rh1 and Rh2 are highly conserved across Old World and New World warblers. However, Rh2 underwent a burst of evolution within the New World genus Setophaga, where it accumulated substitutions at 6 amino acid sites across the species we studied. Evolutionary analyses revealed a significant increase in dN/dS in Setophaga, implying relatively strong selective pressures to overcome long-standing purifying selection. We studied the effects of each substitution on spectral tuning and found they do not cause large spectral shifts. Thus substitutions may reflect other aspects of opsin function, such as those affecting photosensitivity and/or dark-light adaptation. Although it is unclear what these alterations mean for color perception, we suggest that rapid evolution is linked to sexual selection, given the exceptional plumage colour diversification in Setophaga. PMID:25827331

  19. Localization and characterization of val-opsin isoform-expressing cells in the brain of adult zebrafish.

    PubMed

    Hang, Chong Yee; Kitahashi, Takashi; Parhar, Ishwar S

    2014-12-01

    In addition to vision, light information is used to regulate a range of animal physiology. Such nonimage-forming functions of light are mediated by nonvisual photoreceptors expressed in distinct neurons in the retina and the brain in most vertebrates. A nonvisual photoreceptor vertebrate ancient long opsin (VAL-opsin) possesses two functional isoforms in the zebrafish, encoded by valopa and valopb, which has received little attention. To delineate the neurochemical identities of valop cells and to test for colocalization of the valop isoforms, we used in situ hybridization to characterize the expression of the valop genes along with that of neurotransmitters and a neuropeptide known to be present at the sites of valop expression. Double labeling showed that the thalamic valop population coexpresses valopa and valopb. All the thalamic valop cells overlapped with a GABAergic cell mass that continues from the anterior nucleus to the intercalated thalamic nucleus. A novel valopa cell population found in the superior raphe was serotonergic in nature. A valopb cell population in the Edinger-Westphal nucleus was identified as containing thyrotropin-releasing hormone. Valopb cells localized in the hindbrain intermediate reticular formation were noncholinergic in nature (nonmotorneurons). Thus, the presence of valop cell populations in different brain regions with coexpression of neurotransmitters and neuropeptides and the colocalization of valop isoforms in the thalamic cell population indicate regulatory and functional complexity of VAL-opsin in the brain of the zebrafish. PMID:25043553

  20. Characterization of Opsin Gene Alleles Affecting Color Vision in a Wild Population of Titi Monkeys (Callicebus brunneus)

    PubMed Central

    Bunce, John A.; Isbell, Lynne A.; Neitz, Maureen; Bonci, Daniela; Surridge, Alison K.; Jacobs, Gerald H.; Smith, David Glenn

    2011-01-01

    The color vision of most platyrrhine primates is determined by alleles at the polymorphic X-linked locus coding for the opsin responsible for the middle- to long-wavelength (M/L) cone photopigment. Females who are heterozygous at the locus have trichromatic vision while homozygous females and all males are dichromatic. This study characterized the opsin alleles in a wild population of the socially monogamous platyrrhine monkey Callicebus brunneus (the brown titi monkey), a primate that an earlier study suggests may possess an unusual number of alleles at this locus and thus may be a subject of special interest in the study of primate color vision. Direct sequencing of regions of the M/L opsin gene using feces-, blood-, and saliva-derived DNA obtained from 14 individuals yielded evidence for the presence of three functionally distinct alleles, corresponding to the most common M/L photopigment variants inferred from a physiological study of cone spectral sensitivity in captive Callicebus. PMID:20938927

  1. Cloning and expression analysis of two opsin-like genes in the phytopathogenic fungus Bipolaris oryzae.

    PubMed

    Kihara, Junichi; Tanaka, Nozomi; Ueno, Makoto; Arase, Sakae

    2009-06-01

    Two opsin-like genes, OPS1 and OPS2, were identified from a subtracted cDNA library for the identification of near-UV (NUV) radiation-enhanced genes using suppression subtractive hybridization methods in the brown leaf spot fungus Bipolaris oryzae. The OPS1 and OPS2 genes encode predicted proteins of 306 and 304 amino acids, respectively. Real-time PCR analysis showed that the OPS1 transcript is expressed weakly in mycelia under dark conditions but shows enhanced expression after NUV irradiation. By contrast, the OPS2 transcript is constitutively expressed at a high level in mycelia under dark conditions but is only weakly enhanced after NUV irradiation. These enhancement patterns of OPS1 and OPS2 gene expression after NUV irradiation did not occur in the blue-light regulator 1 (BLR1)-deficient mutant, suggesting that NUV radiation-enhanced gene expression of OPS1 and OPS2 could be controlled by the BLR1 in B. oryzae. PMID:19456867

  2. Inhibitory luminopsins: genetically-encoded bioluminescent opsins for versatile, scalable, and hardware-independent optogenetic inhibition

    PubMed Central

    Tung, Jack K.; Gutekunst, Claire-Anne; Gross, Robert E.

    2015-01-01

    Optogenetic techniques provide an unprecedented ability to precisely manipulate neural activity in the context of complex neural circuitry. Although the toolbox of optogenetic probes continues to expand at a rapid pace with more efficient and responsive reagents, hardware-based light delivery is still a major hurdle that limits its practical use in vivo. We have bypassed the challenges of external light delivery by directly coupling a bioluminescent light source (a genetically encoded luciferase) to an inhibitory opsin, which we term an inhibitory luminopsin (iLMO). iLMO was shown to suppress action potential firing and synchronous bursting activity in vitro in response to both external light and luciferase substrate. iLMO was further shown to suppress single-unit firing rate and local field potentials in the hippocampus of anesthetized rats. Finally, expression of iLMO was scaled up to multiple structures of the basal ganglia to modulate rotational behavior of freely moving animals in a hardware-independent fashion. This novel class of optogenetic probes demonstrates how non-invasive inhibition of neural activity can be achieved, which adds to the versatility, scalability, and practicality of optogenetic applications in freely behaving animals. PMID:26399324

  3. The Effect of Cone Opsin Mutations on Retinal Structure and the Integrity of the Photoreceptor Mosaic

    PubMed Central

    Carroll, Joseph; Dubra, Alfredo; Gardner, Jessica C.; Mizrahi-Meissonnier, Liliana; Cooper, Robert F.; Dubis, Adam M.; Nordgren, Rick; Genead, Mohamed; Connor, Thomas B.; Stepien, Kimberly E.; Sharon, Dror; Hunt, David M.; Banin, Eyal; Hardcastle, Alison J.; Moore, Anthony T.; Williams, David R.; Fishman, Gerald; Neitz, Jay; Neitz, Maureen; Michaelides, Michel

    2012-01-01

    Purpose. To evaluate retinal structure and photoreceptor mosaic integrity in subjects with OPN1LW and OPN1MW mutations. Methods. Eleven subjects were recruited, eight of whom have been previously described. Cone and rod density was measured using images of the photoreceptor mosaic obtained from an adaptive optics scanning light ophthalmoscope (AOSLO). Total retinal thickness, inner retinal thickness, and outer nuclear layer plus Henle fiber layer (ONL+HFL) thickness were measured using cross-sectional spectral-domain optical coherence tomography (SD-OCT) images. Molecular genetic analyses were performed to characterize the OPN1LW/OPN1MW gene array. Results. While disruptions in retinal lamination and cone mosaic structure were observed in all subjects, genotype-specific differences were also observed. For example, subjects with “L/M interchange” mutations resulting from intermixing of ancestral OPN1LW and OPN1MW genes had significant residual cone structure in the parafovea (∼25% of normal), despite widespread retinal disruption that included a large foveal lesion and thinning of the parafoveal inner retina. These subjects also reported a later-onset, progressive loss of visual function. In contrast, subjects with the C203R missense mutation presented with congenital blue cone monochromacy, with retinal lamination defects being restricted to the ONL+HFL and the degree of residual cone structure (8% of normal) being consistent with that expected for the S-cone submosaic. Conclusions. The photoreceptor phenotype associated with OPN1LW and OPN1MW mutations is highly variable. These findings have implications for the potential restoration of visual function in subjects with opsin mutations. Our study highlights the importance of high-resolution phenotyping to characterize cellular structure in inherited retinal disease; such information will be critical for selecting patients most likely to respond to therapeutic intervention and for establishing a baseline for evaluating treatment efficacy. PMID:23139274

  4. The halo-opsin gene. II. Sequence, primary structure of halorhodopsin and comparison with bacteriorhodopsin

    PubMed Central

    Blanck, A.; Oesterhelt, D.

    1987-01-01

    The gene for the protein moiety of the light-driven chloride pump halorhodopsin (HR), hop gene, was sequenced and the primary structure of the protein derived thereof. The gene has a GC content of 67% and codes for 274 amino acids. A promoter structure, resembling that of the halobacterial 16S rRNA genes, is present and both a terminating stem and a loop sequence is found downstream of the TGA stop codon. A ribosomal binding site is located within the translated region. The HR protein moiety is processed at the amino terminus, as well as the carboxy terminus, yielding a dominant species of calculated Mr 26 961. Seven transmembrane helical parts of the protein are defined by hydropathy and acrophilicity calculations. Comparision with the bacteriorhodopsin (BR) structure reveals a conservation of 36% of amino acid residues in the transmembrane part and 19% in the connecting loops at both surfaces. The most conspicuous conserved amino acids are the retinal-binding Lys residue, four Trp residues (eventually interacting with retinal), two Asp residues (providing possibly the negative charge environment of retinal) and three Pro residues of unknown function. No significant homology with the opsins of eucaryotes was found. Helical wheel analysis shows that HR is an inside-out protein with the majority of conserved amino acid residues inside the circle of the seven transmembrane helices. It is postulated that the intrahelical spaces, which could be gated by the retinal moiety, are the physical entities for translocation of protons in BR and chloride ions in HR. Retinal, by its cis−trans isomerization, serves as a switch connecting the ion-specific binding sites in both proteins. PMID:15981336

  5. Interspecific variation in Rx1 expression controls opsin expression and causes visual system diversity in African cichlid fishes.

    PubMed

    Schulte, Jane E; O'Brien, Conor S; Conte, Matthew A; O'Quin, Kelly E; Carleton, Karen L

    2014-09-01

    The mechanisms underlying natural phenotypic diversity are key to understanding evolution and speciation. Cichlid fishes are among the most speciose vertebrates and an ideal model for identifying genes controlling species differences. Cichlids have diverse visual sensitivities that result from species expressing subsets of seven cichlid cone opsin genes. We previously identified a quantitative trait locus (QTL) that tunes visual sensitivity by varying SWS2A (short wavelength sensitive 2A) opsin expression in a genetic cross between two Lake Malawi cichlid species. Here, we identify Rx1 (retinal and anterior neural fold homeobox) as the causative gene for the QTL using fine mapping and RNAseq in retinal transcriptomes. Rx1 is differentially expressed between the parental species and correlated with SWS2A expression in the F2 progeny. Expression of Rx1 and SWS2A is also correlated in a panel of 16 Lake Malawi cichlid species. Association mapping in this panel identified a 413-bp deletion located 2.5-kb upstream of the Rx1 translation start site that is correlated with decreased Rx1 expression. This deletion explains 62% of the variance in SWS2A expression across 53 cichlid species in 29 genera. The deletion occurs in both the sand and rock-dwelling cichlid clades, suggesting that it is an ancestral polymorphism. Our finding supports the hypothesis that mixing and matching of ancestral polymorphisms can explain the diversity of present day cichlid phenotypes. PMID:24859246

  6. Possible Involvement of Cone Opsins in Distinct Photoresponses of Intrinsically Photosensitive Dermal Chromatophores in Tilapia Oreochromis niloticus

    PubMed Central

    Chen, Shyh-Chi; Robertson, R. Meldrum; Hawryshyn, Craig W.

    2013-01-01

    Dermal specialized pigment cells (chromatophores) are thought to be one type of extraretinal photoreceptors responsible for a wide variety of sensory tasks, including adjusting body coloration. Unlike the well-studied image-forming function in retinal photoreceptors, direct evidence characterizing the mechanism of chromatophore photoresponses is less understood, particularly at the molecular and cellular levels. In the present study, cone opsin expression was detected in tilapia caudal fin where photosensitive chromatophores exist. Single-cell RT-PCR revealed co-existence of different cone opsins within melanophores and erythrophores. By stimulating cells with six wavelengths ranging from 380 to 580 nm, we found melanophores and erythrophores showed distinct photoresponses. After exposed to light, regardless of wavelength presentation, melanophores dispersed and maintained cell shape in an expansion stage by shuttling pigment granules. Conversely, erythrophores aggregated or dispersed pigment granules when exposed to short- or middle/long-wavelength light, respectively. These results suggest that diverse molecular mechanisms and light-detecting strategies may be employed by different types of tilapia chromatophores, which are instrumental in pigment pattern formation. PMID:23940562

  7. Identification of G protein coupled receptors for opsines and neurohormones in Rhodnius prolixus. Genomic and transcriptomic analysis.

    PubMed

    Ons, Sheila; Lavore, Andrés; Sterkel, Marcos; Wulff, Juan Pedro; Sierra, Ivana; Martínez-Barnetche, Jesús; Rodriguez, Mario Henry; Rivera-Pomar, Rolando

    2016-02-01

    The importance of Chagas disease motivated the scientific effort to obtain the complete genomic sequence of the vector species Rhodnius prolixus, this information is also relevant to the understanding of triatomine biology in general. The central nervous system is the key regulator of insect physiology and behavior. Neurohormones (neuropeptides and biogenic amines) are the chemical messengers involved in the regulation and integration of neuroendocrine signals. In insects, this signaling is mainly mediated by the interaction of neurohormone ligands with G protein coupled receptors (GPCRs). The recently sequenced R. prolixus genome provides us with the opportunity to analyze this important family of genes in triatomines, supplying relevant information for further functional studies. Next-generation sequencing methods offer an excellent opportunity for transcriptomic exploration in key organs and tissues in the presence of a reference genome as well as when a reference genome is not available. We undertook a genomic analysis to obtain a genome-wide inventory of opsines and the GPCRs for neurohormones in R. prolixus. Furthermore, we performed a transcriptomic analysis of R. prolixus central nervous system, focusing on neuropeptide precursor genes and neurohormone and opsines GPCRs. In addition, we mined the whole transcriptomes of Triatoma dimidiata, Triatoma infestans and Triatoma pallidipennis - three sanitary relevant triatomine species - to identify neuropeptide precursors and GPCRs genes. Our study reveals a high degree of sequence conservation in the molecular components of the neuroendocrine system of triatomines. PMID:25976540

  8. X-linked cone dystrophy and colour vision deficiency arising from a missense mutation in a hybrid L/M cone opsin gene

    PubMed Central

    McClements, Michelle; Davies, Wayne I L; Michaelides, Michel; Carroll, Joseph; Rha, Jungate; Mollon, John D; Neitz, Maureen; MacLaren, Robert E; Moore, Anthony T; Hunt, David M

    2013-01-01

    In this report, we describe a male subject who presents with a complex phenotype of myopia associated with cone dysfunction and a protan vision deficiency. Retinal imaging demonstrates extensive cone disruption, including the presence of non-waveguiding cones, an overall thinning of the retina, and an irregular mottled appearance of the hyper reflective band associated with the inner segment ellipsoid portion of the photoreceptor. Mutation screening revealed a novel p.Glu41Lys missense mutation in a hybrid L/M opsin gene. Spectral analysis shows that the mutant opsin fails to form a pigment in vitro and fails to be trafficked to the cell membrane in transfected Neuro2a cells. Extensive sequence and quantitative PCR analysis identifies this mutant gene as the only gene present in the affected subject’s L/M opsin gene array, yet the presence of protanopia indicates that the mutant opsin must retain some activity in vivo. To account for this apparent contradiction, we propose that a limited amount of functional pigment is formed within the normal cellular environment of the intact photoreceptor, and that this requires the presence of chaperone proteins that promote stability and normal folding of the mutant protein. PMID:23337435

  9. Evolution of phototransduction, vertebrate photoreceptors and retina.

    PubMed

    Lamb, Trevor D

    2013-09-01

    Evidence is reviewed from a wide range of studies relevant to the evolution of vertebrate photoreceptors and phototransduction, in order to permit the synthesis of a scenario for the major steps that occurred during the evolution of cones, rods and the vertebrate retina. The ancestral opsin originated more than 700 Mya (million years ago) and duplicated to form three branches before cnidarians diverged from our own lineage. During chordate evolution, ciliary opsins (C-opsins) underwent multiple stages of improvement, giving rise to the 'bleaching' opsins that characterise cones and rods. Prior to the '2R' rounds of whole genome duplication near the base of the vertebrate lineage, 'cone' photoreceptors already existed; they possessed a transduction cascade essentially the same as in modern cones, along with two classes of opsin: SWS and LWS (short- and long-wave-sensitive). These cones appear to have made synaptic contact directly onto ganglion cells, in a two-layered retina that resembled the pineal organ of extant non-mammalian vertebrates. Interestingly, those ganglion cells appear to be descendants of microvillar photoreceptor cells. No lens was associated with this two-layered retina, and it is likely to have mediated circadian timing rather than spatial vision. Subsequently, retinal bipolar cells evolved, as variants of ciliary photoreceptors, and greatly increased the computational power of the retina. With the advent of a lens and extraocular muscles, spatial imaging information became available for central processing, and gave rise to vision in vertebrates more than 500 Mya. The '2R' genome duplications permitted the refinement of cascade components suitable for both rods and cones, and also led to the emergence of five visual opsins. The exact timing of the emergence of 'true rods' is not yet clear, but it may not have occurred until after the divergence of jawed and jawless vertebrates. PMID:23792002

  10. Opsin gene sequence variation across phylogenetic and population histories in Mysis (Crustacea: Mysida) does not match current light environments or visual-pigment absorbance spectra.

    PubMed

    Audzijonyte, Asta; Pahlberg, Johan; Viljanen, Martta; Donner, Kristian; Vinl, Risto

    2012-05-01

    The hypothesis that selection on the opsin gene is efficient in tuning vision to the ambient light environment of an organism was assessed in 49 populations of 12 Mysis crustacean species, inhabiting arctic marine waters, coastal littoral habitats, freshwater lakes ('glacial relicts') and the deep Caspian Sea. Extensive sequence variation was found within and among taxa, but its patterns did not match expectations based on light environments, spectral sensitivity of the visual pigment measured by microspectrophotometry or the history of species and populations. The main split in the opsin gene tree was between lineages I and II, differing in six amino acids. Lineage I was present in marine and Caspian Sea species and in the North American freshwater Mysis diluviana, whereas lineage II was found in the European and circumarctic fresh- and brackish-water Mysis relicta, Mysis salemaai and Mysis segerstralei. Both lineages were present in some populations of M. salemaai and M. segerstralei. Absorbance spectra of the visual pigment in nine populations of the latter three species showed a dichotomy between lake (?(max) =554-562 nm) and brackish-water (Baltic Sea) populations (?(max) = 521-535 nm). Judged by the shape of spectra, this difference was not because of different chromophores (A2 vs. A1), but neither did it coincide with the split in the opsin tree (lineages I/II), species identity or current light environments. In all, adaptive evolution of the opsin gene in Mysis could not be demonstrated, but its sequence variation did not conform to a neutral expectation either, suggesting evolutionary constraints and/or unidentified mechanisms of spectral tuning. PMID:22429275

  11. The influence of L-opsin gene polymorphisms and neural ageing on spatio-chromatic contrast sensitivity in 20-71 year olds.

    PubMed

    Dees, Elise W; Gilson, Stuart J; Neitz, Maureen; Baraas, Rigmor C

    2015-11-01

    Chromatic contrast sensitivity may be a more sensitive measure of an individual's visual function than achromatic contrast sensitivity. Here, the first aim was to quantify individual- and age-related variations in chromatic contrast sensitivity to a range of spatial frequencies for stimuli along two complementary directions in color space. The second aim was to examine whether polymorphisms at specific amino acid residues of the L- and M-opsin genes (OPN1LW and OPN1MW) known to affect spectral tuning of the photoreceptors could influence spatio-chromatic contrast sensitivity. Chromatic contrast sensitivity functions were measured in 50 healthy individuals (20-71 years) employing a novel pseudo-isochromatic grating stimulus. The spatio-chromatic contrast sensitivity functions were found to be low pass for all subjects, independent of age and color vision. The results revealed a senescent decline in spatio-chromatic contrast sensitivity. There were considerable between-individual differences in sensitivity within each age decade for individuals 49 years old or younger, and age did not predict sensitivity for these age decades alone. Forty-six subjects (including a color deficient male and eight female carriers) were genotyped for L- and M-opsin genes. The Ser180Ala polymorphisms on the L-opsin gene were found to influence the subject's color discrimination and their sensitivity to spatio-chromatic patterns. The results expose the significant role of neural and genetic factors in the deterioration of visual function with increasing age. PMID:26368273

  12. Molecular identification and developmental expression of UV and green opsin mRNAs in the pineal organ of the Atlantic halibut.

    PubMed

    Forsell, Johan; Holmqvist, Bo; Ekström, Peter

    2002-05-30

    The pineal organ is the only differentiated photoreceptor organ present in embryos and early larvae of the Atlantic halibut (Hippoglossus hippoglossus). We investigated the molecular identity of opsins in the pineal organ, and their expression during different life stages. Using RT-PCR we identified two 681-bp gene sequences, named HPO1 and HPO4, in cDNA from adult pineal and whole embryos. The predicted amino acid sequences showed highest identity to the transmembrane regions of teleostean RH2 green cone opsins (HPO1, 72-91%) and SWS-1 UV cone opsins (HPO4, 71-83%). In situ hybridization revealed expression of HPO1 and HPO4 mRNA transcripts in photoreceptors in the pineal organ of embryos, larvae and adults. HPO1 and HPO4 mRNA transcripts were also expressed in the larval retina. Our study provides molecular evidence for short and middle wavelength light sensitive photoreceptors in the pineal organ of Atlantic halibut throughout life, and suggests that pineal photoreception may play an important role during embryonic and larval life stages, especially at the time when the retina does not possesses corresponding photoreceptor capacity. PMID:12036517

  13. Opsin expression, physiological characterization and identification of photoreceptor cells in the dorsal rim area and main retina of the desert locust, Schistocerca gregaria.

    PubMed

    Schmeling, Fabian; Wakakuwa, Motohiro; Tegtmeier, Jennifer; Kinoshita, Michiyo; Bockhorst, Tobias; Arikawa, Kentaro; Homberg, Uwe

    2014-10-01

    For compass orientation many insects rely on the pattern of sky polarization, but some species also exploit the sky chromatic contrast. Desert locusts, Schistocerca gregaria, detect polarized light through a specialized dorsal rim area (DRA) in their compound eye. To better understand retinal mechanisms underlying visual navigation, we compared opsin expression, spectral and polarization sensitivities and response-stimulus intensity functions in the DRA and main retina of the locust. In addition to previously characterized opsins of long-wavelength-absorbing (Lo1) and blue-absorbing visual pigments (Lo2), we identified an opsin of an ultraviolet-absorbing visual pigment (LoUV). DRA photoreceptors exclusively expressed Lo2, had peak spectral sensitivities at 441 nm and showed high polarization sensitivity (PS 1.3-31.7). In contrast, ommatidia in the main eye co-expressed Lo1 and Lo2 in five photoreceptors, expressed Lo1 in two proximal photoreceptors, and Lo2 or LoUV in one distal photoreceptor. Correspondingly, we found broadband blue- and green-peaking spectral sensitivities in the main eye and one narrowly tuned UV peaking receptor. Polarization sensitivity in the main retina was low (PS 1.3-3.8). V-log I functions in the DRA were steeper than in the main retina, supporting a role in polarization vision. Desert locusts occur as two morphs, a day-active gregarious and a night-active solitarious form. In solitarious locusts, sensitivities in the main retina were generally shifted to longer wavelengths, particularly in ventral eye regions, supporting a nocturnal lifestyle at low light levels. The data support the role of the DRA in polarization vision and suggest trichromatic colour vision in the desert locust. PMID:25104757

  14. Temporal Resolution of ChR2 and Chronos in an Optogenetic-based Auditory Brainstem Implant Model: Implications for the Development and Application of Auditory Opsins

    PubMed Central

    Hight, A. E.; Kozin, Elliott D.; Darrow, Keith; Lehmann, Ashton; Boyden, Edward; Brown, M. Christian; Lee, Daniel J.

    2015-01-01

    The contemporary auditory brainstem implant (ABI) performance is limited by reliance on electrical stimulation with its accompanying channel cross talk and current spread to non-auditory neurons. A new generation ABI based on optogenetic-technology may ameliorate limitations fundamental to electrical neurostimulation. The most widely studied opsin is channelrhodopsin-2 (ChR2); however, its relatively slow kinetic properties may prevent the encoding of auditory information at high stimulation rates. In the present study, we compare the temporal resolution of light-evoked responses of a recently developed fast opsin, Chronos, to ChR2 in a murine ABI model. Viral mediated gene transfer via a posterolateral craniotomy was used to express Chronos or ChR2 in the mouse nucleus (CN). Following a four to six week incubation period, blue light (473 nm) was delivered via an optical fiber placed directly on the surface of the infected CN, and neural activity was recorded in the contralateral inferior colliculus (IC). Both ChR2 and Chronos evoked sustained responses to all stimuli, even at high driven rates. In addition, optical stimulation evoked excitatory responses throughout the tonotopic axis of the IC. Synchrony of the light-evoked response to stimulus rates of 14–448 pulses/s was higher in Chronos compared to ChR2 mice (p<0.05 at 56, 168, and 224 pulses/s). Our results demonstrate that Chronos has the ability to drive the auditory system at higher stimulation rates than ChR2 and may be a more ideal opsin for manipulation of auditory pathways in future optogenetic-based neuroprostheses. PMID:25598479

  15. Gravitational-wave sensitivity curves

    NASA Astrophysics Data System (ADS)

    Moore, C. J.; Cole, R. H.; Berry, C. P. L.

    2015-01-01

    There are several common conventions in use by the gravitational-wave community to describe the amplitude of sources and the sensitivity of detectors. These are frequently confused. We outline the merits of and differences between the various quantities used for parameterizing noise curves and characterizing gravitational-wave amplitudes. We conclude by producing plots that consistently compare different detectors. Similar figures can be generated on-line for general use at http://rhcole.com/apps/GWplotter.

  16. Co-translational targeting and translocation of the amino terminus of opsin across the endoplasmic membrane requires GTP but not ATP.

    PubMed

    Kanner, Elliott M; Friedlander, Martin; Simon, Sanford M

    2003-03-01

    The tight coupling between ongoing translation and translocation across the mammalian endoplasmic reticulum has made it difficult to determine the requirements that are specific for translocation. We have developed an in vitro assay that faithfully mimics the co-translational targeting and translocation of the amino terminus of opsin without ongoing translation. Using this system we demonstrate that this post-translational targeting and translocation requires nucleotide triphosphates but not cytosolic proteins. The addition of GTP alone was sufficient to fully restore targeting. The addition of ATP was not specifically required, and non-hydrolyzable analogs of ATP that blocked 90% of the ATPase activity also had no inhibitory effect on translocation. PMID:12486130

  17. Opsin1-2, Gqα and arrestin levels at Limulus rhabdoms are controlled by diurnal light and a circadian clock

    PubMed Central

    Battelle, Barbara-Anne; Kempler, Karen E.; Parker, Alexander K.; Gaddie, Cristina D.

    2013-01-01

    SUMMARY Dark and light adaptation in photoreceptors involve multiple processes including those that change protein concentrations at photosensitive membranes. Light- and dark-adaptive changes in protein levels at rhabdoms have been described in detail in white-eyed Drosophila maintained under artificial light. Here we tested whether protein levels at rhabdoms change significantly in the highly pigmented lateral eyes of wild-caught Limulus polyphemus maintained in natural diurnal illumination and whether these changes are under circadian control. We found that rhabdomeral levels of opsins (Ops1-2), the G protein activated by rhodopsin (Gqα) and arrestin change significantly from day to night and that nighttime levels of each protein at rhabdoms are significantly influenced by signals from the animal's central circadian clock. Clock input at night increases Ops1-2 and Gqα and decreases arrestin levels at rhabdoms. Clock input is also required for a rapid decrease in rhabdomeral Ops1-2 beginning at sunrise. We found further that dark adaptation during the day and the night are not equivalent. During daytime dark adaptation, when clock input is silent, the increase of Ops1-2 at rhabdoms is small and Gqα levels do not increase. However, increases in Ops1-2 and Gqα at rhabdoms are enhanced during daytime dark adaptation by treatments that elevate cAMP in photoreceptors, suggesting that the clock influences dark-adaptive increases in Ops1-2 and Gqα at Limulus rhabdoms by activating cAMP-dependent processes. The circadian regulation of Ops1-2 and Gqα levels at rhabdoms probably has a dual role: to increase retinal sensitivity at night and to protect photoreceptors from light damage during the day. PMID:23393287

  18. ISO Guest Observer Data Analysis and LWS Instrument Team Activities

    NASA Technical Reports Server (NTRS)

    Oliversen, Ronald J. (Technical Monitor); Smith, Howard A.

    2003-01-01

    We have designed and fabricated infrared filters for use at wavelengths greater than or equal to 15 microns. Unlike conventional dielectric filters used at the short wavelengths, ours are made from stacked metal grids, spaced at a very small fraction of the performance wavelengths. The individual lattice layers are gold, the spacers are polyimide, and they are assembled using integrated circuit processing techniques; they resemble some metallic photonic band-gap structures. We simulate the filter performance accurately, including the coupling of the propagating, near-field electromagnetic modes, using computer aided design codes. We find no anomalous absorption. The geometrical parameters of the grids are easily altered in practice, allowing for the production of tuned filters with predictable useful transmission characteristics. Although developed for astronomical instrumentation, the filters are broadly applicable in systems across infrared and terahertz bands.

  19. ISO Guest Observer Data Analysis and LWS Instrument Team Activities

    NASA Technical Reports Server (NTRS)

    Smith, Howard

    2002-01-01

    This project was granted a no-cost extension prompted by the request of the major subcontractor, the Naval Research Laboratory, which had not yet completed its tasks. As of July 2002, they had made substantial progress. They have successfully fabricated a metal mesh grid on polyimide, and also successfully fabricated a 2-layer metal mesh infrared filter using stacks of these metal mesh grids on polyimide; the actual layering was done at SAO. Both warm and cold spectroscopic tests were done on these fabricated devices. The measurements were in good agreement with the theory, and also reasonable performance in absolute terms. NRL is now working on fabricating a 3-layer metal mesh infrared filter, and a prototype is expected in the next month. Testing should occur before the end of the fiscal year. Finally, NRL has preliminarily agreed to hire a new postdoctoral person to refine the modeling of the filters based on the new measurements. The person should arrive this fall. NRL has a new Fourier Transform Spectrometer which will be delivered in the next month, and which will be used to facilitate the testing which has up to now been done in collaboration with NASA Goddard Space Flight Space Center.

  20. ISO Guest Observer Data Analysis and LWS Instrument Team Activities

    NASA Technical Reports Server (NTRS)

    Smith, Howard

    2001-01-01

    The following is an interim annual report. Dr. Smith is currently on an extended TDY to the Istituto di Fisica dello Spazio Interplanetario (IFSI) at the Consilio Nazionale delle Richerche (CNR) in Rome, Italy, where he has been working on a related NASA grant in support of analysis of Infrared Space Observatory (ISO) data on star formation in Ultra Luminous Infrared Galaxies and our galaxy. Work emphasizes development of metal mesh grids for use in spacecraft, and the design and fabrication of test elements by the Naval Research Laboratory, Washington D.C. Work has progressed well, but slowly, on that program due to the departure of a key engineer. NASA has been advised of the delay, and granted a no-cost extension, whereby SAO has authorized a delay in the final report from NRL. Nevertheless NRL has continued to make progress. Two papers have been submitted to refereed journals related to this program, and a new design for mesh operating in the 20-40 micron region has been developed. Meetings continue through the summer on these items. A new technical scientist has been made a job offer and hopefully will be on board NRL shortly, although most of the present grant work is already completed. A more complete report, with copies of the submitted papers, designs, and other measures of progress, will be submitted to NASA in September when Dr. Smith returns from his current TDY.

  1. Extending the LWS Data Environment: Distributed Data Processing and Analysis

    NASA Technical Reports Server (NTRS)

    Narock, Thomas

    2005-01-01

    The final stages of this work saw changes to the original framework, as well as the completion and integration of several data processing services. Initially, it was thought that a peer-to-peer architecture was necessary to make this work possible. The peer-to-peer architecture provided many benefits including the dynamic discovery of new services that would be continually added. A prototype example was built and while it showed promise, a major disadvantage was seen in that it was not easily integrated into the existing data environment. While the peer-to-peer system worked well for finding and accessing distributed data processing services, it was found that its use was limited by the difficulty in calling it from existing tools and services. After collaborations with members of the data community, it was determined that our data processing system was of high value and that a new interface should be pursued in order for the community to take full advantage of it. As such; the framework was modified from a peer-to-peer architecture to a more traditional web service approach. Following this change multiple data processing services were added. These services include such things as coordinate transformations and sub setting of data. Observatory (VHO), assisted with integrating the new architecture into the VHO. This allows anyone using the VHO to search for data, to then pass that data through our processing services prior to downloading it. As a second attempt at demonstrating the new system, a collaboration was established with the Collaborative Sun Earth Connector (CoSEC) group at Lockheed Martin. This group is working on a graphical user interface to the Virtual Observatories and data processing software. The intent is to provide a high-level easy-to-use graphical interface that will allow access to the existing Virtual Observatories and data processing services from one convenient application. Working with the CoSEC group we provided access to our data processing tools from within their software. This now allows the CoSEC community to take advantage of our services and also demonstrates another means of accessing our system.

  2. Targeting of exon VI-skipping human RGR-opsin to the plasma membrane of pigment epithelium and co-localization with terminal complement complex C5b-9

    PubMed Central

    Kochounian, Harold; Zhang, Zhaoxia; Spee, Christine; Hinton, David R.

    2016-01-01

    Purpose Rare mutations in the human RGR gene lead to autosomal recessive retinitis pigmentosa or dominantly inherited peripapillary choroidal atrophy. Here, we analyze a common exon-skipping isoform of the human retinal G protein-coupled receptor opsin (RGR-d) to determine differences in subcellular targeting between RGR-d and normal RGR and possible association with abnormal traits in the human eye. Methods The terminal complement complex (C5b-9), vitronectin, CD46, syntaxin-4, and RGR-d were analyzed in human eye tissue from young and old donors or in cultured fetal RPE cells by means of immunofluorescent labeling and high-resolution confocal microscopy or immunohistochemical staining. Results We observed that RGR-d is targeted to the basolateral plasma membrane of the RPE. RGR-d, but not normal RGR, is expressed in cultured human fetal RPE cells in which the protein also trafficks to the plasma membrane. In young donors, the amount of RGR-d protein in the basolateral plasma membrane was much higher than that in the RPE cells of older subjects. In older donor eyes, the level of immunoreactive RGR-d within RPE cells was often low or undetectable, and immunostaining of RGR-d was consistently strongest in extracellular deposits in Bruch’s membrane. Double immunofluorescent labeling in the basal deposits revealed significant aggregate and small punctate co-localization of RGR-d with C5b-9 and vitronectin. Conclusions RGR-d may escape endoplasmic reticulum-associated degradation and in contrast to full-length RGR, traffick to the basolateral plasma membrane, particularly in younger subjects. RGR-d in the plasma membrane indicates that the protein is properly folded, as misfolded membrane proteins cannot otherwise sort to the plasma membrane. The close association of extracellular RGR-d with both vitronectin and C5b-9 suggests a potential role of RGR-d-containing deposits in complement activation. PMID:27011730

  3. A novel molecular marker for the study of Neotropical cichlid phylogeny.

    PubMed

    Fabrin, T M C; Gasques, L S; Prioli, S M A P; Prioli, A J

    2015-01-01

    The use of molecular markers has contributed to phylogeny and to the reconstruction of species' evolutionary history. Each region of the genome has different evolution rates, which may or may not identify phylogenetic signal at different levels. Therefore, it is important to assess new molecular markers that can be used for phylogenetic reconstruction. Regions that may be associated with species characteristics and are subject to selective pressure, such as opsin genes, which encode proteins related to the visual system and are widely expressed by Cichlidae family members, are interesting. Our aim was to identify a new nuclear molecular marker that could establish the phylogeny of Neotropical cichlids and is potentially correlated with the visual system. We used Bayesian inference and maximum likelihood analysis to support the use of the nuclear opsin LWS gene in the phylogeny of eight Neotropical cichlid species. Their use concatenated to the mitochondrial gene COI was also tested. The LWS gene fragment comprised the exon 2-4 region, including the introns. The LWS gene provided good support for both analyses up to the genus level, distinguishing the studied species, and when concatenated to the COI gene, there was a good support up to the species level. Another benefit of utilizing this region, is that some polymorphisms are associated with changes in spectral properties of the LWS opsin protein, which constitutes the visual pigment that absorbs red light. Thus, utilization of this gene as a molecular marker to study the phylogeny of Neotropical cichlids is promising. PMID:26782460

  4. Multiple Genetic Mechanisms Contribute to Visual Sensitivity Variation in the Labridae.

    PubMed

    Phillips, Genevieve A C; Carleton, Karen L; Marshall, N Justin

    2016-01-01

    Coral reefs are one of the most spectrally diverse environments, both in terms of habitat and animal color. Species identity, sex, and camouflage are drivers of the phenotypic diversity seen in coral reef fishes, but how the phenotypic diversity is reflected in the genotype remains to be answered. The labrids are a large, polyphyletic family of coral reef fishes that display a diverse range of colors, including developmental color morphs and extensive behavioral ecologies. Here, we assess the opsin sequence and expression diversity among labrids from the Great Barrier Reef, Australia. We found that labrids express a diverse palette of visual opsins, with gene duplications in both RH2 and LWS genes. The majority of opsins expressed were within the mid-to-long wavelength sensitive classes (RH2 and LWS). Three of the labrid species expressed SWS1 (ultra-violet sensitive) opsins with the majority expressing the violet-sensitive SWS2B gene and none expressing SWS2A. We used knowledge about spectral tuning sites to calculate approximate spectral sensitivities (λmax) for individual species' visual pigments, which corresponded well with previously published λmax values for closely related species (SWS1: 356-370 nm; SWS2B: 421-451 nm; RH2B: 452-492 nm; RH2A: 516-528 nm; LWS1: 554-555 nm; LWS2: 561-562 nm). In contrast to the phenotypic diversity displayed via color patterns and feeding ecology, there was little amino acid diversity within the known opsin sequence tuning sites. However, gene duplications and differential expression provide alternative mechanisms for tuning visual pigments, resulting in variable visual sensitivities among labrid species. PMID:26464127

  5. Visual system evolution and the nature of the ancestral snake.

    PubMed

    Simões, B F; Sampaio, F L; Jared, C; Antoniazzi, M M; Loew, E R; Bowmaker, J K; Rodriguez, A; Hart, N S; Hunt, D M; Partridge, J C; Gower, D J

    2015-07-01

    The dominant hypothesis for the evolutionary origin of snakes from 'lizards' (non-snake squamates) is that stem snakes acquired many snake features while passing through a profound burrowing (fossorial) phase. To investigate this, we examined the visual pigments and their encoding opsin genes in a range of squamate reptiles, focusing on fossorial lizards and snakes. We sequenced opsin transcripts isolated from retinal cDNA and used microspectrophotometry to measure directly the spectral absorbance of the photoreceptor visual pigments in a subset of samples. In snakes, but not lizards, dedicated fossoriality (as in Scolecophidia and the alethinophidian Anilius scytale) corresponds with loss of all visual opsins other than RH1 (λmax 490-497 nm); all other snakes (including less dedicated burrowers) also have functional sws1 and lws opsin genes. In contrast, the retinas of all lizards sampled, even highly fossorial amphisbaenians with reduced eyes, express functional lws, sws1, sws2 and rh1 genes, and most also express rh2 (i.e. they express all five of the visual opsin genes present in the ancestral vertebrate). Our evidence of visual pigment complements suggests that the visual system of stem snakes was partly reduced, with two (RH2 and SWS2) of the ancestral vertebrate visual pigments being eliminated, but that this did not extend to the extreme additional loss of SWS1 and LWS that subsequently occurred (probably independently) in highly fossorial extant scolecophidians and A. scytale. We therefore consider it unlikely that the ancestral snake was as fossorial as extant scolecophidians, whether or not the latter are para- or monophyletic. PMID:26012745

  6. Cone visual pigments of monotremes: filling the phylogenetic gap.

    PubMed

    Wakefield, Matthew J; Anderson, Mark; Chang, Ellen; Wei, Ke-Jun; Kaul, Rajinder; Graves, Jennifer A Marshall; Grützner, Frank; Deeb, Samir S

    2008-01-01

    We have determined the sequence and genomic organization of the genes encoding the cone visual pigment of the platypus (Ornithorhynchus anatinus) and the echidna (Tachyglossus aculeatus), and inferred their spectral properties and evolutionary pathways. We prepared platypus and echidna retinal RNA and used primers of the middle-wave-sensitive (MWS), long-wave-sensitive (LWS), and short-wave sensitive (SWS1) pigments corresponding to coding sequences that are highly conserved among mammals; to PCR amplify the corresponding pigment sequences. Amplification from the retinal RNA revealed the expression of LWS pigment mRNA that is homologous in sequence and spectral properties to the primate LWS visual pigments. However, we were unable to amplify the mammalian SWS1 pigment from these two species, indicating this gene was lost prior to the echidna-platypus divergence (21 MYA). Subsequently, when the platypus genome sequence became available, we found an LWS pigment gene in a conserved genomic arrangement that resembles the primate pigment, but, surprisingly we found an adjacent (20 kb) SWS2 pigment gene within this conserved genomic arrangement. We obtained the same result after sequencing the echidna genes. The encoded SWS2 pigment is predicted to have a wavelength of maximal absorption of about 440 nm, and is paralogous to SWS pigments typically found in reptiles, birds, and fish but not in mammals. This study suggests the locus control region (LCR) has played an important role in the conservation of photo receptor gene arrays and the control of their spatial and temporal expression in the retina in all mammals. In conclusion, a duplication event of an ancestral cone visual pigment gene, followed by sequence divergence and selection gave rise to the LWS and SWS2 visual pigments. So far, the echidna and platypus are the only mammals that share the gene structure of the LWS-SWS2 pigment gene complex with reptiles, birds and fishes. PMID:18598396

  7. Beauty in the eyes of the beholders: colour vision is tuned to mate preference in the Trinidadian guppy (Poecilia reticulata).

    PubMed

    Sandkam, Benjamin; Young, C Megan; Breden, Felix

    2015-02-01

    A broad range of animals use visual signals to assess potential mates, and the theory of sensory exploitation suggests variation in visual systems drives mate preference variation due to sensory bias. Trinidadian guppies (Poecilia reticulata), a classic system for studies of the evolution of female mate choice, provide a unique opportunity to test this theory by looking for covariation in visual tuning, light environment and mate preferences. Female preference co-evolves with male coloration, such that guppy females from 'low-predation' environments have stronger preferences for males with more orange/red coloration than do females from 'high-predation' environments. Here, we show that colour vision also varies across populations, with 'low'-predation guppies investing more of their colour vision to detect red/orange coloration. In independently colonized watersheds, guppies expressed higher levels of both LWS-1 and LWS-3 (the most abundant LWS opsins) in 'low-predation' populations than 'high-predation' populations at a time that corresponds to differences in cone cell abundance. We also observed that the frequency of a coding polymorphism differed between high- and low-predation populations. Together, this shows that the variation underlying preference could be explained by simple changes in expression and coding of opsins, providing important candidate genes to investigate the genetic basis of female preference variation in this model system. PMID:25556876

  8. Cone visual pigments of aquatic mammals.

    PubMed

    Newman, Lucy A; Robinson, Phyllis R

    2005-01-01

    It has long been hypothesized that the visual systems of animals are evolutionarily adapted to their visual environment. The entrance many millions of years ago of mammals into the sea gave these new aquatic mammals completely novel visual surroundings with respect to light availability and predominant wavelengths. This study examines the cone opsins of marine mammals, hypothesizing, based on previous studies [Fasick et al. (1998) and Levenson & Dizon (2003)], that the deep-dwelling marine mammals would not have color vision because the pressure to maintain color vision in the dark monochromatic ocean environment has been relaxed. Short-wavelength-sensitive (SWS) and long-wavelength-sensitive (LWS) cone opsin genes from two orders (Cetacea and Sirenia) and an additional suborder (Pinnipedia) of aquatic mammals were amplified from genomic DNA (for SWS) and cDNA (for LWS) by PCR, cloned, and sequenced. All animals studied from the order Cetacea have SWS pseudogenes, whereas a representative from the order Sirenia has an intact SWS gene, for which the corresponding mRNA was found in the retina. One of the pinnipeds studied (harp seal) has an SWS pseudogene, while another species (harbor seal) appeared to have an intact SWS gene. However, no SWS cone opsin mRNA was found in the harbor seal retina, suggesting a promoter or splice site mutation preventing transcription of the gene. The LWS opsins from the different species were expressed in mammalian cells and reconstituted with the 11-cis-retinal chromophore in order to determine maximal absorption wavelengths (lambda(max)) for each. The deeper dwelling Cetacean species had blue shifted lambda(max) values compared to shallower-dwelling aquatic species. Taken together, these findings support the hypothesis that in the monochromatic oceanic habitat, the pressure to maintain color vision has been relaxed and mutations are retained in the SWS genes, resulting in pseudogenes. Additionally, LWS opsins are retained in the retina and, in deeper-dwelling animals, are blue shifted in lambda(max). PMID:16469194

  9. LWS Proposal to Provide Scientific Guidance and Modeling Support for the Ionospheric Mapping Mission. Part 1

    NASA Technical Reports Server (NTRS)

    Richmond, Arthur D.

    2005-01-01

    A data assimilation system for specifying the thermospheric density has been developed over the last several years. This system ingests GRACE/CHAMP-type in situ as well as SSULI/SSUSI remote sensing observations while making use of a physical model, the Coupled Thermosphere-Ionosphere Model (CTIM) (Fuller-Rowel1 et al., 1996). The Kalman filter was implemented as the backbone to the data assimilation system, which provides a statistically 'best' estimate as well as an estimate of the error in its state. The system was tested using a simulated thermosphere and observations. CHAMP data were then used to provide the system with a real data source. The results of this study are herein.

  10. 2006 LWS TR & T Solar Wind Focused Science Topic Team: The Beginnings

    NASA Astrophysics Data System (ADS)

    Miralles, M. P.; Bhattacharjee, A.; Landi, E.; Markovskii, S.; Cranmer, S. R.; Doschek, G. A.; Forbes, T. G.; Isenberg, P. A.; Kohl, J. L.; Ng, C.; Raymond, J. C.; Vasquez, B. J.

    2006-12-01

    The Solar Wind Focused Science Topic (FST) team was created to apply a combination of theoretical studies, numerical simulations, and observations to the understanding of how the fast and slow solar wind are heated and accelerated. Four proposals were selected for this FST team. They will investigate the role of energy sources and kinetic mechanisms responsible for the heating and acceleration of the solar wind. In particular, the FST team will examine magnetic reconnection and turbulence as possible heating mechanisms. Plasma properties and their evolution over the solar cycle, determined from the analysis of remote and in situ measurements, will be used to put firm constraints on the models. The work of the Solar Wind FST team is in its initial stages. The organization, planning, and findings resulting from the first FST team meeting will be reported.

  11. Flight Experiments for Living With a Star Space Environment Testbed (LWS-SET): Relationship to Technology

    NASA Technical Reports Server (NTRS)

    LaBel, Kenneth A.; Barth, Janet L.; Brewer, Dana A.

    2003-01-01

    This viewgraph presentation provides information on flight validation experiments for technologies to determine solar effects. The experiments are intended to demonstrate tolerance to a solar variant environment. The technologies tested are microelectronics, photonics, materials, and sensors.

  12. The LWS Geospace Storm Investigations Exploring the Extremes of Space Weather

    NASA Technical Reports Server (NTRS)

    2002-01-01

    The Geospace mission of the Living With a Star program is a family of investigations focusing on the compelling science questions that advance our ability to specify, understand, and predict the societal impact of solar variance. Two key areas have been identified as combining both importance to society and potential for scientific progress: 1) characterization and understanding of the acceleration, global distribution, and variability of energetic electrons and ions in the inner magnetosphere, and 2) characterization and understanding of the ionosphere and irregularities that affect communications, navigation and radar systems. Under these broad categories specific science questions have emerged as the priority science objectives for the first Geospace Investigations: How and why do relativistic electrons in the outer zone and slot region vary during geomagnetic storms? How does the long- and short-term variability of the Sun affect the global-scale behavior of the ionospheric electron density and irregularities, especially during magnetic storms and at mid-latitudes? The first Geospace mission will attempt to answer these questions.

  13. Demonstration of a genotype-phenotype correlation in the polymorphic color vision of a non-callitrichine New World monkey, capuchin (Cebus apella).

    PubMed

    Saito, Atsuko; Kawamura, Shoji; Mikami, Akichika; Ueno, Yoshikazu; Hiramatsu, Chihiro; Koida, Kowa; Fujita, Kazuo; Kuroshima, Hika; Hasegawa, Toshikazu

    2005-12-01

    Color-vision polymorphism in New World monkeys occurs because of an allelic polymorphism of the single-copy red-green middle-to-long-wavelength-sensitive (M/LWS) opsin gene on the X chromosome. Because color-vision types can readily be estimated from allelic types of the M/LWS opsin gene, this polymorphic system offers researchers an excellent opportunity to study the association between vision and behavior. As a prerequisite for such studies, genetically determined color-vision types must be concordant with phenotypes determined directly by behavioral criteria (e.g., by a color discrimination test). However, such correlations between genotypes and phenotypes have been studied only for callitrichine species. Using genetic, electrophysiological, and behavioral approaches, we evaluated the color vision of brown capuchin monkeys (Cebus apella), a representative non-callitrichine model animal for physiology and behavior. Two allelic M/LWS opsins-P545 and P530-were identified in the studied captive population. Females had one or both of the alleles, and males had either one. The retinal sensitivity in P530 dichromats was short-wave shifted relative to that in P545 dichromats, whereas that in P530/P545 trichromats was between the two groups. In a discrimination task using Ishihara pseudo-isochromatic plates, P530/P545 trichromats were successful in discriminating stimuli that P530 and P545 dichromats were unable to discriminate. In a food-search task, P530/P545 trichromats were able to locate red targets among green distracters as quickly as among white distracters, whereas both types of dichromats took longer. These results demonstrate the mutual consistency between genotypes and phenotypes of color vision, and provide a solid genetic basis on which the ecology and evolution of color vision can be investigated. PMID:16342070

  14. Cone photoreceptors and potential UV vision in a subterranean insectivore, the European mole.

    PubMed

    Glösmann, Martin; Steiner, Marianne; Peichl, Leo; Ahnelt, Peter K

    2008-01-01

    We have examined the presence, the distribution, and the opsin identity of photoreceptor types in the retina of the European mole, Talpa europaea, a subterranean insectivore with regressed morphology of the visual system. Cones and rods were identified using opsin antisera, and their topographies determined from flat-mounted retinas. The retina (total area 0.75 mm(2)) contains about 100,000 photoreceptors, 10-12% of which are cones. Rod density is low (theoretical maximum 127,000 mm(-2)). Cone density peaks in central retina (17,750 mm(-2)). Similar to most mammals, two cone opsins, shortwave-sensitive (S) and middle-to-long-wave-sensitive (M), are present. Cone distribution shows a dorsoventral gradient with higher S cone numbers in ventral retina. Coexpression of S and M opsin occurs in more than 30% of the cones. Partial sequencing of the S opsin gene strongly supports UV sensitivity of the mole S cone photopigment. Amino acids that spectrally tune the S opsin are identical in T. europaea and in mammals with known UV cone photosensitivity. The lens transmits light down to 300 nm. Together, our data suggest that photopic vision and UV sensitivity of a cone pigment play a functional role in the European mole. PMID:18484862

  15. Cone monochromacy and visual pigment spectral tuning in wobbegong sharks

    PubMed Central

    Theiss, Susan M.; Davies, Wayne I. L.; Collin, Shaun P.; Hunt, David M.; Hart, Nathan S.

    2012-01-01

    Much is known regarding the evolution of colour vision in nearly every vertebrate class, with the notable exception of the elasmobranchs. While multiple spectrally distinct cone types are found in some rays, sharks appear to possess only a single class of cone and, therefore, may be colour blind. In this study, the visual opsin genes of two wobbegong species, Orectolobus maculatus and Orectolobus ornatus, were isolated to verify the molecular basis of their monochromacy. In both species, only two opsin genes are present, RH1 (rod) and LWS (cone), which provide further evidence to support the concept that sharks possess only a single cone type. Examination of the coding sequences revealed substitutions that account for interspecific variation in the photopigment absorbance spectra, which may reflect the difference in visual ecology between these species. PMID:22993239

  16. Cone monochromacy and visual pigment spectral tuning in wobbegong sharks.

    PubMed

    Theiss, Susan M; Davies, Wayne I L; Collin, Shaun P; Hunt, David M; Hart, Nathan S

    2012-12-23

    Much is known regarding the evolution of colour vision in nearly every vertebrate class, with the notable exception of the elasmobranchs. While multiple spectrally distinct cone types are found in some rays, sharks appear to possess only a single class of cone and, therefore, may be colour blind. In this study, the visual opsin genes of two wobbegong species, Orectolobus maculatus and Orectolobus ornatus, were isolated to verify the molecular basis of their monochromacy. In both species, only two opsin genes are present, RH1 (rod) and LWS (cone), which provide further evidence to support the concept that sharks possess only a single cone type. Examination of the coding sequences revealed substitutions that account for interspecific variation in the photopigment absorbance spectra, which may reflect the difference in visual ecology between these species. PMID:22993239

  17. Effects of exogenous thyroid hormones on visual pigment composition in coho salmon (Oncorhynchus kisutch).

    PubMed

    Temple, Shelby E; Ramsden, Samuel D; Haimberger, Theodore J; Veldhoen, Kathy M; Veldhoen, Nik J; Carter, Nicolette L; Roth, Wolff-Michael; Hawryshyn, Craig W

    2008-07-01

    The role of exogenous thyroid hormone on visual pigment content of rod and cone photoreceptors was investigated in coho salmon (Oncorhynchus kisutch). Coho vary the ratio of vitamin A1- and A2-based visual pigments in their eyes. This variability potentially alters spectral sensitivity and thermal stability of the visual pigments. We tested whether the direction of shift in the vitamin A1/A2 ratio, resulting from application of exogenous thyroid hormone, varied in fish of different ages and held under different environmental conditions. Changes in the vitamin A1/A2 visual pigment ratio were estimated by measuring the change in maximum absorbance (lambda max) of rods using microspectrophotometry (MSP). Exogenous thyroid hormone resulted in a long-wavelength shift in rod, middle-wavelength-sensitive (MWS) and long-wavelength-sensitive (LWS) cone photoreceptors. Rod and LWS cone lambda max values increased, consistent with an increase in vitamin A2. MWS cone lambda max values increased more than predicted for a change in the vitamin A1/A2 ratio. To account for this shift, we tested for the expression of multiple RH2 opsin subtypes. We isolated and sequenced a novel RH2 opsin subtype, which had 48 amino acid differences from the previously sequenced coho RH2 opsin. A substitution of glutamate for glutamine at position 122 could partially account for the greater than predicted shift in MWS cone lambda max values. Our findings fit the hypothesis that a variable vitamin A1/A2 ratio provides seasonality in spectral tuning and/or improved thermal stability of visual pigments in the face of seasonal environmental changes, and that multiple RH2 opsin subtypes can provide flexibility in spectral tuning associated with migration-metamorphic events. PMID:18552303

  18. The Transcription Factor GTF2IRD1 Regulates the Topology and Function of Photoreceptors by Modulating Photoreceptor Gene Expression across the Retina

    PubMed Central

    Masuda, Tomohiro; Zhang, Xiaodong; Berlinicke, Cindy; Wan, Jun; Yerrabelli, Anitha; Conner, Elizabeth A.; Kjellstrom, Sten; Bush, Ronald; Thorgeirsson, Snorri S.; Swaroop, Anand; Chen, Shiming

    2014-01-01

    The mechanisms that specify photoreceptor cell-fate determination, especially as regards to short-wave-sensitive (S) versus medium-wave-sensitive (M) cone identity, and maintain their nature and function, are not fully understood. Here we report the importance of general transcription factor II-I repeat domain-containing protein 1 (GTF2IRD1) in maintaining M cone cell identity and function as well as rod function. In the mouse, GTF2IRD1 is expressed in cell-fate determined photoreceptors at postnatal day 10. GTF2IRD1 binds to enhancer and promoter regions in the mouse rhodopsin, M- and S-opsin genes, but regulates their expression differentially. Through interaction with the transcription factors CRX and thyroid hormone receptor β 2, it enhances M-opsin expression, whereas it suppresses S-opsin expression; and with CRX and NRL, it enhances rhodopsin expression. In an apparent paradox, although GTF2IRD1 is widely expressed in multiple cell types across the retina, knock-out of GTF2IRD1 alters the retinal expression of only a limited number of annotated genes. Interestingly, however, the null mutation leads to altered topology of cone opsin expression in the retina, with aberrant S-opsin overexpression and M-opsin underexpression in M cones. Gtf2ird1-null mice also demonstrate abnormal M cone and rod electrophysiological responses. These findings suggest an important role for GTF2IRD1 in regulating the level and topology of rod and cone gene expression, and in maintaining normal retinal function. PMID:25392503

  19. Modelling rock-avalanche induced impact waves: Sensitivity of the model chains to model parameters

    NASA Astrophysics Data System (ADS)

    Schaub, Yvonne; Huggel, Christian

    2014-05-01

    New lakes are forming in high-mountain areas all over the world due to glacier recession. Often they will be located below steep, destabilized flanks and are therefore exposed to impacts from rock-/ice-avalanches. Several events worldwide are known, where an outburst flood has been triggered by such an impact. In regions such as in the European Alps or in the Cordillera Blanca in Peru, where valley bottoms are densely populated, these far-travelling, high-magnitude events can result in major disasters. Usually natural hazards are assessed as single hazardous processes, for the above mentioned reasons, however, development of assessment and reproduction methods of the hazardous process chain for the purpose of hazard map generation have to be brought forward. A combination of physical process models have already been suggested and illustrated by means of lake outburst in the Cordillera Blanca, Peru, where on April 11th 2010 an ice-avalanche of approx. 300'000m3 triggered an impact wave, which overtopped the 22m freeboard of the rock-dam for 5 meters and caused and outburst flood which travelled 23 km to the city of Carhuaz. We here present a study, where we assessed the sensitivity of the model chain from ice-avalanche and impact wave to single parameters considering rock-/ice-avalanche modeling by RAMMS and impact wave modeling by IBER. Assumptions on the initial rock-/ice-avalanche volume, calibration of the friction parameters in RAMMS and assumptions on erosion considered in RAMMS were parameters tested regarding their influence on overtopping parameters that are crucial for outburst flood modeling. Further the transformation of the RAMMS-output (flow height and flow velocities on the shoreline of the lake) into an inflow-hydrograph for IBER was also considered a possible source of uncertainties. Overtopping time, volume, and wave height as much as mean and maximum discharge were considered decisive parameters for the outburst flood modeling and were therewith assumed dependent values. The resulting 54 runs were evaluated by an ANOVA-analysis for each dependent variable. Results show, that the model chain is able to correctly reproduce the 5m-overtopping wave. Further the dependency from the input parameters could be assessed for every dependent variable. It was e.g. shown, that RAMMS-calibration has the strongest influence on all variations, it is more crucial then the uncertainties introduced by assumptions on the initial rock-avalanche volume. The study shows, that from a hazard-assessment point of view, combinations of model chains are acceptable and permissible.

  20. Developmental plasticity in vision and behavior may help guppies overcome increased turbidity.

    PubMed

    Ehlman, Sean M; Sandkam, Benjamin A; Breden, Felix; Sih, Andrew

    2015-12-01

    Increasing turbidity in streams and rivers near human activity is cause for environmental concern, as the ability of aquatic organisms to use visual information declines. To investigate how some organisms might be able to developmentally compensate for increasing turbidity, we reared guppies (Poecilia reticulata) in either clear or turbid water. We assessed the effects of developmental treatments on adult behavior and aspects of the visual system by testing fish from both developmental treatments in turbid and clear water. We found a strong interactive effect of rearing and assay conditions: fish reared in clear water tended to decrease activity in turbid water, whereas fish reared in turbid water tended to increase activity in turbid water. Guppies from all treatments decreased activity when exposed to a predator. To measure plasticity in the visual system, we quantified treatment differences in opsin gene expression of individuals. We detected a shift from mid-wave-sensitive opsins to long wave-sensitive opsins for guppies reared in turbid water. Since long-wavelength sensitivity is important in motion detection, this shift likely allows guppies to salvage motion-detecting abilities when visual information is obscured in turbid water. Our results demonstrate the importance of developmental plasticity in responses of organisms to rapidly changing environments. PMID:26427995

  1. The activation of directional stem cell motility by green light-emitting diode irradiation.

    PubMed

    Ong, Wei-Kee; Chen, How-Foo; Tsai, Cheng-Ting; Fu, Yun-Ju; Wong, Yi-Shan; Yen, Da-Jen; Chang, Tzu-Hao; Huang, Hsien-Da; Lee, Oscar Kuang-Sheng; Chien, Shu; Ho, Jennifer Hui-Chun

    2013-03-01

    Light-emitting diode (LED) irradiation is potentially a photostimulator to manipulate cell behavior by opsin-triggered phototransduction and thermal energy supply in living cells. Directional stem cell motility is critical for the efficiency and specificity of stem cells in tissue repair. We explored that green LED (530 nm) irradiation directed the human orbital fat stem cells (OFSCs) to migrate away from the LED light source through activation of extracellular signal-regulated kinases (ERK)/MAP kinase/p38 signaling pathway. ERK inhibitor selectively abrogated light-driven OFSC migration. Phosphorylation of these kinases as well as green LED irradiation-induced cell migration was facilitated by increasing adenosine triphosphate (ATP) production in OFSCs after green LED exposure, and which was thermal stress-independent mechanism. OFSCs, which are multi-potent mesenchymal stem cells isolated from human orbital fat tissue, constitutionally express three opsins, i.e. retinal pigment epithelium-derived rhodopsin homolog (RRH), encephalopsin (OPN3) and short-wave-sensitive opsin 1 (OPN1SW). However, only two non-visual opsins, i.e. RRH and OPN3, served as photoreceptors response to green LED irradiation-induced OFSC migration. In conclusion, stem cells are sensitive to green LED irradiation-induced directional cell migration through activation of ERK signaling pathway via a wavelength-dependent phototransduction. PMID:23261211

  2. Early evolution of vertebrate photoreception: lessons from lampreys and lungfishes.

    PubMed

    Collin, Shaun P

    2009-03-01

    Lampreys (Agnatha) and lungfish (Dipnoi) are representatives of the earliest and the intermediate stages in vertebrate evolution, respectively, and survived in the Cambrian (approximately 540 mA, lampreys) and Devonian (approximately 400 mA, lungfishes) Periods. The unique phylogenetic position of these two groups presents us with an exciting opportunity to understand life in ancient times and to begin to trace the evolution of vision and photoreception in vertebrates. Using a multidisciplinary approach employing anatomical and molecular techniques, the evolution of photoreception is explored in these extant, living fossils to predict the environmental lighting conditions to which our vertebrate ancestors were exposed. Contrary to expectations, the retinae of the southern hemisphere lamprey (Geotria australis Gray, 1851) and the Australian lungfish (Neoceratodus forsteri Krefft, 1870) are far from "primitive," each possessing five types of photoreceptors, many with spectral filters for tuning the light. Detailed ultrastructural analysis reveals that all five receptor types in G. australis are cone-like, whereas N. forsteri possesses four cone types and a single type of rod. Each receptor type also contains a different visual pigment (opsin gene); that is, LWS, SWS1, SWS2, RhA and RhB in G. australis and LWS, SWS1, SWS2, Rh1 and Rh2 in N. forsteri, all of which are expressed within the retina and are sensitive to different parts of the electromagnetic spectrum, providing the potential for pentachromatic and tetrachromatic color vision, respectively. PMID:21392279

  3. Evolution of the vertebrate eye: opsins, photoreceptors, retina and eye cup

    PubMed Central

    Lamb, Trevor D.; Collin, Shaun P.; Pugh, Edward N.

    2011-01-01

    Charles Darwin appreciated the conceptual difficulty in accepting that an organ as wonderful as the vertebrate eye could have evolved through natural selection. He reasoned that if appropriate gradations could be found that were useful to the animal and were inherited, then the apparent difficulty would be overcome. Here, we review a wide range of findings that capture glimpses of the gradations that appear to have occurred during eye evolution, and provide a scenario for the unseen steps that have led to the emergence of the vertebrate eye. PMID:18026166

  4. Opsin switch reveals function of the ultraviolet cone in fish foraging

    PubMed Central

    Novales Flamarique, Iñigo

    2013-01-01

    Although several studies have shown that ultraviolet (UV) wavelengths are important in naturally occurring, visually guided behaviours of vertebrates, the function of the UV cone in such behaviours is unknown. Here, I used thyroid hormone to transform the UV cones of young rainbow trout into blue cones, a phenomenon that occurs naturally as the animal grows, to test whether the resulting loss of UV sensitivity affected the animal's foraging performance on Daphnia magna, a prey zooplankton. The distances and angles at which prey were located (variables that are known indicators of foraging performance) were significantly reduced for UV knock-out fish compared with controls. Optical measurements and photon-catch calculations revealed that the contrast of Daphnia was greater when perceived by the visual system of control versus that of thyroid-hormone-treated fish, demonstrating that the UV cone enhanced the foraging performance of young rainbow trout. Because most juvenile fishes have UV cones and feed on zooplankton, this finding has wide implications for understanding the visual ecology of fishes. The enhanced target contrast provided by UV cones could be used by other vertebrates in various behaviours, including foraging, mate selection and communication. PMID:23222448

  5. Short-wavelength sensitive opsin (SWS1) as a new marker for vertebrate phylogenetics

    PubMed Central

    van Hazel, Ilke; Santini, Francesco; Müller, Johannes; Chang, Belinda SW

    2006-01-01

    Background Vertebrate SWS1 visual pigments mediate visual transduction in response to light at short wavelengths. Due to their importance in vision, SWS1 genes have been isolated from a surprisingly wide range of vertebrates, including lampreys, teleosts, amphibians, reptiles, birds, and mammals. The SWS1 genes exhibit many of the characteristics of genes typically targeted for phylogenetic analyses. This study investigates both the utility of SWS1 as a marker for inferring vertebrate phylogenetic relationships, and the characteristics of the gene that contribute to its phylogenetic utility. Results Phylogenetic analyses of vertebrate SWS1 genes produced topologies that were remarkably congruent with generally accepted hypotheses of vertebrate evolution at both higher and lower taxonomic levels. The few exceptions were generally associated with areas of poor taxonomic sampling, or relationships that have been difficult to resolve using other molecular markers. The SWS1 data set was characterized by a substantial amount of among-site rate variation, and a relatively unskewed substitution rate matrix, even when the data were partitioned into different codon sites and individual taxonomic groups. Although there were nucleotide biases in some groups at third positions, these biases were not convergent across different taxonomic groups. Conclusion Our results suggest that SWS1 may be a good marker for vertebrate phylogenetics due to the variable yet consistent patterns of sequence evolution exhibited across fairly wide taxonomic groups. This may result from constraints imposed by the functional role of SWS1 pigments in visual transduction. PMID:17107620

  6. Of Mice, Men, and Microbial Opsins: How Optogenetics Can Help Hone Mouse Models of Mental Illness.

    PubMed

    Marton, Tobias F; Sohal, Vikaas S

    2016-01-01

    Genetic, pharmacologic, and behavioral manipulations have long been powerful tools for generating rodent models to study the neural substrates underlying psychiatric disease. Recent advances in the use of optogenetics in awake behaving rodents has added an additional valuable methodology to this experimental toolkit. Here, we review several recent studies that leverage optogenetic technologies to elucidate neural mechanisms possibly related to depression, anxiety, and obsessive-compulsive disorder. We use a few illustrative examples to highlight key emergent principles about how optogenetics, in conjunction with more established modalities, can help to organize our understanding of how disease-related states, specific neuronal circuits, and various behavioral assays fit into hierarchical frameworks such as the National Institute of Mental Health Research Domain Criteria matrix. PMID:25981174

  7. Gene conversion between red and defective green opsin gene in blue cone monochromacy

    SciTech Connect

    Reyniers, E.; Van Thienen, M.N.; De Boulle, K.; Willems, P.J.

    1995-09-20

    Blue cone monochromacy is an X-linked condition in which the function of both the red pigment gene (RCP) and the green pigment gene (GCP) is impaired. Blue cone monochromacy can be due to a red/green gene array rearrangement existing of a single red/green hybrid gene and an inactivating C203R point mutation in both RCP and GCP. The flanking sequences of the C230R mutation in exon 4 of RCP were characteristic for GCP, indicating that this mutation was transferred from GCP into RCP by gene conversion. 23 refs., 3 figs., 1 tab.

  8. Temperature profile analysis for Amargosa Valley wells LWS-A, ASH-B, and MSH-C. DOE UGTA RI/FS geothermal gradient study results FY 1995

    SciTech Connect

    Gillespie, D.

    1995-09-01

    The purpose of this investigation is to estimate vertical fluid flux rates in saturated units penetrated by selected wells at the Nevada Test Site (NTS) using geothermal gradient data. Estimates of vertical fluid flux rates are critical in developing models for contaminate flow within, recharge potential to, and discharge from the groundwater system at the NTS. If temperature in the well is in equilibrium with the surrounding rocks, it is possible to detect the vertical flow of groundwater from the well`s thermal profile. Heat in the subsurface is transported by conduction through the rock and by advection caused by subsurface water movement. Units in which vertical flow is occurring will produce a curve in the thermal profile within the well. Prior to the implementation of the Underground Test Area Remedial Investigation/Feasibility Study (UGTA RI/FS), investigation of thermal data from wells at the NTS was hindered by the completion of wells as open holes or without casing cemented in place. The open-hole type of completion allows cross-flow within the wellbore which can yield information about relative pressures between connected aquifers, but which renders thermal gradient data essentially useless, or at best highly suspect for interpreting in situ groundwater movement. Wells recently completed in the Department of Energy (DOE) Environmental Restoration (ER) Program have been completed with casing cemented to the surface (to prohibit cross-flow between units in the annular space between the casing and the wellbore) and with completion zones open to a single hydrologic horizon. This type of completion results in temperature gradient profiles more representative of actual thermal conditions in the units penetrated by the well. Results are presented of temperature profiles of 3 wells located in the Death Valley Groundwater Basin, outside the southern border of the NTS.

  9. MODELING LARGE WOOD STRUCTURES IN SAND BED STREAMS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In-stream large wood structures (LWS) are becoming increasingly popular throughout the world. The LWS improve aquatic habitat quality and protect banks from erosion. While most reports describe the LWS in the Northwest as successful, LWS in one Mississippi sand-bed stream had an unacceptable failure...

  10. Eye spectral sensitivity in fresh- and brackish-water populations of three glacial-relict Mysis species (Crustacea): physiology and genetics of differential tuning.

    PubMed

    Donner, Kristian; Zak, Pavel; Viljanen, Martta; Lindström, Magnus; Feldman, Tatiana; Ostrovsky, Mikhail

    2016-04-01

    Absorbance spectra of single rhabdoms were studied by microspectrophotometry (MSP) and spectral sensitivities of whole eyes by electroretinography (ERG) in three glacial-relict species of opossum shrimps (Mysis). Among eight populations from Fennoscandian fresh-water lakes (L) and seven populations from the brackish-water Baltic Sea (S), L spectra were systematically red-shifted by 20-30 nm compared with S spectra, save for one L and one S population. The difference holds across species and bears no consistent adaptive relation to the current light environments. In the most extensively studied L-S pair, two populations of M. relicta (L p and S p) separated for less than 10,000 years, no differences translating into amino acid substitutions have been found in the opsin genes, and the chromophore of the visual pigments as analyzed by HPLC is pure A1. However, MSP experiments with spectrally selective bleaching show the presence of two rhodopsins (λ max ≈ 525-530 nm, MWS, and 565-570 nm, LWS) expressed in different proportions. ERG recordings of responses to "red" and "blue" light linearly polarized at orthogonal angles indicate segregation of the pigments into different cells differing in polarization sensitivity. We propose that the pattern of development of LWS and MWS photoreceptors is governed by an ontogenetic switch responsive to some environmental signal(s) other than light that generally differ(s) between lakes and sea, and that this reaction norm is conserved from a common ancestor of all three species. PMID:26984686

  11. Spatial And Temporal Aspects Of Red/Green Opponency

    NASA Astrophysics Data System (ADS)

    Kelly, D. H.

    1988-06-01

    Like other aspects of color vision, interactions between the visual pathways of the long-wave-sensitive (LWS) and the middle-wave-sensitive (MWS) photoreceptors can no longer be regarded only as functions of wavelength. The results of color matching, detection, discrimination and cancellation procedures may also vary with spatial and/or temporal properties of the stimulus. In some ways, the spatial and temporal stimulus dimensions have analogous effects on color perception. For example, the (opponent-color) responses to isoluminant, red/green, sine-wave stimuli are low-pass in both spatial and temporal frequency domains, whereas the (luminous) responses to monochromatic or achromatic stimuli emphasize higher frequencies in both domains. But spatial patterns can have properties that temporal waveforms cannot, involving orientation or symmetry.Thus the simplest model for spatial phase relations--a linear filter with zero phase-shift--is forbidden in the temporal case. This paper emphasizes the properties of 3-way (spatiotemporal-chromatic) interactions, and discusses the role of separability in modelling these interactions.

  12. RELAX: detecting relaxed selection in a phylogenetic framework.

    PubMed

    Wertheim, Joel O; Murrell, Ben; Smith, Martin D; Kosakovsky Pond, Sergei L; Scheffler, Konrad

    2015-03-01

    Relaxation of selective strength, manifested as a reduction in the efficiency or intensity of natural selection, can drive evolutionary innovation and presage lineage extinction or loss of function. Mechanisms through which selection can be relaxed range from the removal of an existing selective constraint to a reduction in effective population size. Standard methods for estimating the strength and extent of purifying or positive selection from molecular sequence data are not suitable for detecting relaxed selection, because they lack power and can mistake an increase in the intensity of positive selection for relaxation of both purifying and positive selection. Here, we present a general hypothesis testing framework (RELAX) for detecting relaxed selection in a codon-based phylogenetic framework. Given two subsets of branches in a phylogeny, RELAX can determine whether selective strength was relaxed or intensified in one of these subsets relative to the other. We establish the validity of our test via simulations and show that it can distinguish between increased positive selection and a relaxation of selective strength. We also demonstrate the power of RELAX in a variety of biological scenarios where relaxation of selection has been hypothesized or demonstrated previously. We find that obligate and facultative γ-proteobacteria endosymbionts of insects are under relaxed selection compared with their free-living relatives and obligate endosymbionts are under relaxed selection compared with facultative endosymbionts. Selective strength is also relaxed in asexual Daphnia pulex lineages, compared with sexual lineages. Endogenous, nonfunctional, bornavirus-like elements are found to be under relaxed selection compared with exogenous Borna viruses. Finally, selection on the short-wavelength sensitive, SWS1, opsin genes in echolocating and nonecholocating bats is relaxed only in lineages in which this gene underwent pseudogenization; however, selection on the functional medium/long-wavelength sensitive opsin, M/LWS1, is found to be relaxed in all echolocating bats compared with nonecholocating bats. PMID:25540451

  13. RELAX: Detecting Relaxed Selection in a Phylogenetic Framework

    PubMed Central

    Wertheim, Joel O.; Murrell, Ben; Smith, Martin D.; Kosakovsky Pond, Sergei L.; Scheffler, Konrad

    2015-01-01

    Relaxation of selective strength, manifested as a reduction in the efficiency or intensity of natural selection, can drive evolutionary innovation and presage lineage extinction or loss of function. Mechanisms through which selection can be relaxed range from the removal of an existing selective constraint to a reduction in effective population size. Standard methods for estimating the strength and extent of purifying or positive selection from molecular sequence data are not suitable for detecting relaxed selection, because they lack power and can mistake an increase in the intensity of positive selection for relaxation of both purifying and positive selection. Here, we present a general hypothesis testing framework (RELAX) for detecting relaxed selection in a codon-based phylogenetic framework. Given two subsets of branches in a phylogeny, RELAX can determine whether selective strength was relaxed or intensified in one of these subsets relative to the other. We establish the validity of our test via simulations and show that it can distinguish between increased positive selection and a relaxation of selective strength. We also demonstrate the power of RELAX in a variety of biological scenarios where relaxation of selection has been hypothesized or demonstrated previously. We find that obligate and facultative γ-proteobacteria endosymbionts of insects are under relaxed selection compared with their free-living relatives and obligate endosymbionts are under relaxed selection compared with facultative endosymbionts. Selective strength is also relaxed in asexual Daphnia pulex lineages, compared with sexual lineages. Endogenous, nonfunctional, bornavirus-like elements are found to be under relaxed selection compared with exogenous Borna viruses. Finally, selection on the short-wavelength sensitive, SWS1, opsin genes in echolocating and nonecholocating bats is relaxed only in lineages in which this gene underwent pseudogenization; however, selection on the functional medium/long-wavelength sensitive opsin, M/LWS1, is found to be relaxed in all echolocating bats compared with nonecholocating bats. PMID:25540451

  14. From the ultrasonic to the infrared: molecular evolution and the sensory biology of bats.

    PubMed

    Jones, Gareth; Teeling, Emma C; Rossiter, Stephen J

    2013-01-01

    Great advances have been made recently in understanding the genetic basis of the sensory biology of bats. Research has focused on the molecular evolution of candidate sensory genes, genes with known functions [e.g., olfactory receptor (OR) genes] and genes identified from mutations associated with sensory deficits (e.g., blindness and deafness). For example, the FoxP2 gene, underpinning vocal behavior and sensorimotor coordination, has undergone diversification in bats, while several genes associated with audition show parallel amino acid substitutions in unrelated lineages of echolocating bats and, in some cases, in echolocating dolphins, representing a classic case of convergent molecular evolution. Vision genes encoding the photopigments rhodopsin and the long-wave sensitive opsin are functional in bats, while that encoding the short-wave sensitive opsin has lost functionality in rhinolophoid bats using high-duty cycle laryngeal echolocation, suggesting a sensory trade-off between investment in vision and echolocation. In terms of olfaction, bats appear to have a distinctive OR repertoire compared with other mammals, and a gene involved in signal transduction in the vomeronasal system has become non-functional in most bat species. Bitter taste receptors appear to have undergone a "birth-and death" evolution involving extensive gene duplication and loss, unlike genes coding for sweet and umami tastes that show conservation across most lineages but loss in vampire bats. Common vampire bats have also undergone adaptations for thermoperception, via alternative splicing resulting in the evolution of a novel heat-sensitive channel. The future for understanding the molecular basis of sensory biology is promising, with great potential for comparative genomic analyses, studies on gene regulation and expression, exploration of the role of alternative splicing in the generation of proteomic diversity, and linking genetic mechanisms to behavioral consequences. PMID:23755015

  15. From the ultrasonic to the infrared: molecular evolution and the sensory biology of bats

    PubMed Central

    Jones, Gareth; Teeling, Emma C.; Rossiter, Stephen J.

    2013-01-01

    Great advances have been made recently in understanding the genetic basis of the sensory biology of bats. Research has focused on the molecular evolution of candidate sensory genes, genes with known functions [e.g., olfactory receptor (OR) genes] and genes identified from mutations associated with sensory deficits (e.g., blindness and deafness). For example, the FoxP2 gene, underpinning vocal behavior and sensorimotor coordination, has undergone diversification in bats, while several genes associated with audition show parallel amino acid substitutions in unrelated lineages of echolocating bats and, in some cases, in echolocating dolphins, representing a classic case of convergent molecular evolution. Vision genes encoding the photopigments rhodopsin and the long-wave sensitive opsin are functional in bats, while that encoding the short-wave sensitive opsin has lost functionality in rhinolophoid bats using high-duty cycle laryngeal echolocation, suggesting a sensory trade-off between investment in vision and echolocation. In terms of olfaction, bats appear to have a distinctive OR repertoire compared with other mammals, and a gene involved in signal transduction in the vomeronasal system has become non-functional in most bat species. Bitter taste receptors appear to have undergone a “birth-and death” evolution involving extensive gene duplication and loss, unlike genes coding for sweet and umami tastes that show conservation across most lineages but loss in vampire bats. Common vampire bats have also undergone adaptations for thermoperception, via alternative splicing resulting in the evolution of a novel heat-sensitive channel. The future for understanding the molecular basis of sensory biology is promising, with great potential for comparative genomic analyses, studies on gene regulation and expression, exploration of the role of alternative splicing in the generation of proteomic diversity, and linking genetic mechanisms to behavioral consequences. PMID:23755015

  16. Visual pigments in a living fossil, the Australian lungfish Neoceratodus forsteri

    PubMed Central

    Bailes, Helena J; Davies, Wayne L; Trezise, Ann EO; Collin, Shaun P

    2007-01-01

    Background One of the greatest challenges facing the early land vertebrates was the need to effectively interpret a terrestrial environment. Interpretation was based on ocular adaptations evolved for an aquatic environment millions of years earlier. The Australian lungfish Neoceratodus forsteri is thought to be the closest living relative to the first terrestrial vertebrate, and yet nothing is known about the visual pigments present in lungfish or the early tetrapods. Results Here we identify and characterise five visual pigments (rh1, rh2, lws, sws1 and sws2) expressed in the retina of N. forsteri. Phylogenetic analysis of the molecular evolution of lungfish and other vertebrate visual pigment genes indicates a closer relationship between lungfish and amphibian pigments than to pigments in teleost fishes. However, the relationship between lungfish, the coelacanth and tetrapods could not be absolutely determined from opsin phylogeny, supporting an unresolved trichotomy between the three groups. Conclusion The presence of four cone pigments in Australian lungfish suggests that the earliest tetrapods would have had a colorful view of their terrestrial environment. PMID:17961206

  17. Advantage of dichromats over trichromats in discrimination of color-camouflaged stimuli in nonhuman primates.

    PubMed

    Saito, Atsuko; Mikami, Akichika; Kawamura, Shoji; Ueno, Yoshikazu; Hiramatsu, Chihiro; Widayati, Kanthi A; Suryobroto, Bambang; Teramoto, Migaku; Mori, Yusuke; Nagano, Kunitoshi; Fujita, Kazuo; Kuroshima, Hika; Hasegawa, Toshikazu

    2005-12-01

    Due to a middle- to long-wavelength-sensitive (M/LWS) cone opsin polymorphism, there is considerable phenotypic variation in the color vision of New World monkeys. Many females have trichromatic vision, whereas some females and all males have dichromatic vision. The selective pressures that maintain this polymorphism are unclear. In the present study we compared the performance of dichromats and trichromats in a discrimination task. We examined tri- and dichromatic individuals of two species: brown capuchin monkeys (Cebus apella) and long-tailed macaques (Macaca fascicularis). We also examined one protanomalous chimpanzee (Pan troglodytes). The subjects' task was to discriminate a circular pattern from other patterns in which textural elements differed in orientation and thickness from the background. After they were trained with stimuli of a single color, the subjects were presented with color-camouflaged stimuli with a green/red mosaic overlaid onto the pattern. The dichromatic monkeys and the protanomalous chimpanzee selected the correct stimulus under camouflaged conditions at rates significantly above chance levels, while the trichromats did not. These findings demonstrate that dichromatic nonhuman primates possess a superior visual ability to discriminate color-camouflaged stimuli, and that such an ability may confer selective advantages with respect to the detection of cryptic foods and/or predators. PMID:16342068

  18. Optogenetic manipulation of cGMP in cells and animals by the tightly light-regulated guanylyl-cyclase opsin CyclOp.

    PubMed

    Gao, Shiqiang; Nagpal, Jatin; Schneider, Martin W; Kozjak-Pavlovic, Vera; Nagel, Georg; Gottschalk, Alexander

    2015-01-01

    Cyclic GMP (cGMP) signalling regulates multiple biological functions through activation of protein kinase G and cyclic nucleotide-gated (CNG) channels. In sensory neurons, cGMP permits signal modulation, amplification and encoding, before depolarization. Here we implement a guanylyl cyclase rhodopsin from Blastocladiella emersonii as a new optogenetic tool (BeCyclOp), enabling rapid light-triggered cGMP increase in heterologous cells (Xenopus oocytes, HEK293T cells) and in Caenorhabditis elegans. Among five different fungal CyclOps, exhibiting unusual eight transmembrane topologies and cytosolic N-termini, BeCyclOp is the superior optogenetic tool (light/dark activity ratio: 5,000; no cAMP production; turnover (20 °C) ∼17 cGMP s(-1)). Via co-expressed CNG channels (OLF in oocytes, TAX-2/4 in C. elegans muscle), BeCyclOp photoactivation induces a rapid conductance increase and depolarization at very low light intensities. In O2/CO2 sensory neurons of C. elegans, BeCyclOp activation evokes behavioural responses consistent with their normal sensory function. BeCyclOp therefore enables precise and rapid optogenetic manipulation of cGMP levels in cells and animals. PMID:26345128

  19. Optogenetic manipulation of cGMP in cells and animals by the tightly light-regulated guanylyl-cyclase opsin CyclOp

    PubMed Central

    Gao, Shiqiang; Nagpal, Jatin; Schneider, Martin W.; Kozjak-Pavlovic, Vera; Nagel, Georg; Gottschalk, Alexander

    2015-01-01

    Cyclic GMP (cGMP) signalling regulates multiple biological functions through activation of protein kinase G and cyclic nucleotide-gated (CNG) channels. In sensory neurons, cGMP permits signal modulation, amplification and encoding, before depolarization. Here we implement a guanylyl cyclase rhodopsin from Blastocladiella emersonii as a new optogenetic tool (BeCyclOp), enabling rapid light-triggered cGMP increase in heterologous cells (Xenopus oocytes, HEK293T cells) and in Caenorhabditis elegans. Among five different fungal CyclOps, exhibiting unusual eight transmembrane topologies and cytosolic N-termini, BeCyclOp is the superior optogenetic tool (light/dark activity ratio: 5,000; no cAMP production; turnover (20 °C) ∼17 cGMP s−1). Via co-expressed CNG channels (OLF in oocytes, TAX-2/4 in C. elegans muscle), BeCyclOp photoactivation induces a rapid conductance increase and depolarization at very low light intensities. In O2/CO2 sensory neurons of C. elegans, BeCyclOp activation evokes behavioural responses consistent with their normal sensory function. BeCyclOp therefore enables precise and rapid optogenetic manipulation of cGMP levels in cells and animals. PMID:26345128

  20. Age-related deterioration of rod vision in mice.

    PubMed

    Kolesnikov, Alexander V; Fan, Jie; Crouch, Rosalie K; Kefalov, Vladimir J

    2010-08-18

    Even in healthy individuals, aging leads to deterioration in visual acuity, contrast sensitivity, visual field, and dark adaptation. Little is known about the neural mechanisms that drive the age-related changes of the retina and, more specifically, photoreceptors. According to one hypothesis, the age-related deterioration in rod function is due to the limited availability of 11-cis-retinal for rod pigment formation. To determine how aging affects rod photoreceptors and to test the retinoid-deficiency hypothesis, we compared the morphological and functional properties of rods of adult and aged B6D2F1/J mice. We found that the number of rods and the length of their outer segments were significantly reduced in 2.5-year-old mice compared with 4-month-old animals. Aging also resulted in a twofold reduction in the total level of opsin in the retina. Behavioral tests revealed that scotopic visual acuity and contrast sensitivity were decreased by twofold in aged mice, and rod ERG recordings demonstrated reduced amplitudes of both a- and b-waves. Sensitivity of aged rods determined from single-cell recordings was also decreased by 1.5-fold, corresponding to not more than 1% free opsin in these photoreceptors, and kinetic parameters of dim flash response were not altered. Notably, the rate of rod dark adaptation was unaffected by age. Thus, our results argue against age-related deficiency of 11-cis-retinal in the B6D2F1/J mouse rod visual cycle. Surprisingly, the level of cellular dark noise was increased in aged rods, providing an alternative mechanism for their desensitization. PMID:20720130

  1. Evolutionary changes of multiple visual pigment genes in the complete genome of Pacific bluefin tuna.

    PubMed

    Nakamura, Yoji; Mori, Kazuki; Saitoh, Kenji; Oshima, Kenshiro; Mekuchi, Miyuki; Sugaya, Takuma; Shigenobu, Yuya; Ojima, Nobuhiko; Muta, Shigeru; Fujiwara, Atushi; Yasuike, Motoshige; Oohara, Ichiro; Hirakawa, Hideki; Chowdhury, Vishwajit Sur; Kobayashi, Takanori; Nakajima, Kazuhiro; Sano, Motohiko; Wada, Tokio; Tashiro, Kosuke; Ikeo, Kazuho; Hattori, Masahira; Kuhara, Satoru; Gojobori, Takashi; Inouye, Kiyoshi

    2013-07-01

    Tunas are migratory fishes in offshore habitats and top predators with unique features. Despite their ecological importance and high market values, the open-ocean lifestyle of tuna, in which effective sensing systems such as color vision are required for capture of prey, has been poorly understood. To elucidate the genetic and evolutionary basis of optic adaptation of tuna, we determined the genome sequence of the Pacific bluefin tuna (Thunnus orientalis), using next-generation sequencing technology. A total of 26,433 protein-coding genes were predicted from 16,802 assembled scaffolds. From these, we identified five common fish visual pigment genes: red-sensitive (middle/long-wavelength sensitive; M/LWS), UV-sensitive (short-wavelength sensitive 1; SWS1), blue-sensitive (SWS2), rhodopsin (RH1), and green-sensitive (RH2) opsin genes. Sequence comparison revealed that tuna's RH1 gene has an amino acid substitution that causes a short-wave shift in the absorption spectrum (i.e., blue shift). Pacific bluefin tuna has at least five RH2 paralogs, the most among studied fishes; four of the proteins encoded may be tuned to blue light at the amino acid level. Moreover, phylogenetic analysis suggested that gene conversions have occurred in each of the SWS2 and RH2 loci in a short period. Thus, Pacific bluefin tuna has undergone evolutionary changes in three genes (RH1, RH2, and SWS2), which may have contributed to detecting blue-green contrast and measuring the distance to prey in the blue-pelagic ocean. These findings provide basic information on behavioral traits of predatory fish and, thereby, could help to improve the technology to culture such fish in captivity for resource management. PMID:23781100

  2. Design of Large Wood Structures in Sand-Bed Streams

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Large woody structures (LWS) are potentially an efficient and cost effective way to protect streambanks from erosion while enhancing aquatic habitat. While LWS have been successful in some cases in the Pacific Northwest when ballasted with rock, the failure rate in sand-bed streams typical of the mi...

  3. Photochemical Nature of Parietopsin

    PubMed Central

    Sakai, Kazumi; Imamoto, Yasushi; Su, Chih-Ying; Tsukamoto, Hisao; Yamashita, Takahiro; Terakita, Akihisa; Yau, King-Wai; Shichida, Yoshinori

    2012-01-01

    Parietopsin is a non-visual green-light-sensitive opsin closely related to vertebrate visual opsins, and was originally identified in lizard parietal-eye photoreceptor cells. To obtain insight into the functional diversity of opsins, we investigated by UV-visible absorption spectroscopy the molecular properties of parietopsin and its mutants exogenously expressed in cultured cells, and compared to vertebrate and invertebrate visual opsins. Our mutational analysis revealed that the counterion in parietopsin is the glutamic acid (Glu) in the second extracellular loop, corresponding to Glu181 in bovine rhodopsin. This arrangement is characteristic of invertebrate rather than vertebrate visual opsins. The photosensitivity and the molar extinction coefficient of parietopsin were also lower than those of vertebrate visual opsins, features likewise characteristic of invertebrate visual opsins. On the other hand, irradiation of parietopsin yielded meta-I, meta-II, and meta-III intermediates after batho- and lumi-intermediates, similar to vertebrate visual opsins. The pH-dependent equilibrium profile between meta-I and meta-II intermediates was, however, similar to that between acid and alkaline metarhodopsins in invertebrate visual opsins. Thus, parietopsin behaves as an “evolutionary intermediate” between invertebrate and vertebrate visual opsins. PMID:22303823

  4. Developmental dynamics of cone photoreceptors in the eel

    PubMed Central

    2009-01-01

    Background Many fish alter their expressed visual pigments during development. The number of retinal opsins expressed and their type is normally related to the environment in which they live. Eels are known to change the expression of their rod opsins as they mature, but might they also change the expression of their cone opsins? Results The Rh2 and Sws2 opsin sequences from the European Eel were isolated, sequenced and expressed in vitro for an accurate measurement of their λmax values. In situ hybridisation revealed that glass eels express only rh2 opsin in their cone photoreceptors, while larger yellow eels continue to express rh2 opsin in the majority of their cones, but also have <5% of cones which express sws2 opsin. Silver eels showed the same expression pattern as the larger yellow eels. This observation was confirmed by qPCR (quantitative polymerase chain reaction). Conclusions Larger yellow and silver European eels express two different cone opsins, rh2 and sws2. This work demonstrates that only the Rh2 cone opsin is present in younger fish (smaller yellow and glass), the sws2 opsin being expressed additionally only by older fish and only in <5% of cone cells. PMID:20025774

  5. Differential gene expression in mouse retina related to regional differences in vulnerability to hyperoxia

    PubMed Central

    Natoli, Riccardo; Valter, Krisztina; Stone, Jonathan

    2010-01-01

    Purpose In the C57BL/6J mouse retina, hyperoxia-induced degeneration of photoreceptors shows strong regional variation, beginning at a locus ~0.5 mm inferior to the optic disc. To identify gene expression differences that might underlie this variability in vulnerability, we have used microarray techniques to describe regional (superior-inferior) variations in gene expression in the retina. Methods Young adult C57BL/6J mice raised in dim cyclic illumination (12 h at 5 lx and 12 h in darkness) were exposed to hyperoxia (75% oxygen for two weeks). Retinas were collected from hyperoxia-exposed and control animals without fixation and divided into superior and inferior halves. RNA was extracted from each sample, purified, and hybridized to Mouse Gene 1.0 ST arrays (Affymetrix). The consistency of the microarray results was assessed using quantitative PCR for selected genes. Expression data were analyzed to identify genes and ncRNAs whose differential expression between the superior and inferior retina could be associated with relative vulnerability to hyperoxia. Results In control retinas, only two genes showed a fold difference in expression >2 between the superior and inferior retina; another 25 showed a fold difference of 1.5–2.0. Of these 27, the functions of six genes, including ventral anterior homeobox containing gene 2 (Vax2) and T-box 5 (Tbox5), are related to parameters of anatomic development and the functions of five are related to sensory perception. Among the latter, short-wave-sensitive cone opsin (Opn1sw) was more strongly expressed in the inferior retina and medium-wave-sensitive cone opsin (Opn1mw) in the superior retina. This is consistent with known differences in S- and M-cone distribution, confirming our separation of retinal regions. The highest fold difference was reported for membrane metalloendopeptidase (Mme), a member from the metallothionein group of cytoprotective proteins. To identify genes whose regulation by hyperoxia was significantly different between the inferior and superior retina, we calculated the “fold margin” (FM, the difference between hyperoxia-induced regulation in the inferior and superior retina) for each gene, and identified genes for which abs(FM) > 0.5. Genes thus identified numbered 112, and included many immune-, cell defense-, and inflammation- related genes. Conclusions Gene expression analysis revealed relatively subtle differences between inferior and superior regions of control C57BL/6J retinas, with only 27 genes showing an expression difference >1.5 fold. Among these, genes related to cytoprotection and apoptosis were included, along with genes related to central projections and cone-type differences. After hyperoxia-induced photoreceptor degeneration had begun, the number of genes that showed significant expression differences between the inferior and superior retina more than quadrupled, with genes related to immune processes, defense processes, and inflammation being numerically dominant. PMID:20454693

  6. The nop-1 gene of Neurospora crassa encodes a seven transmembrane helix retinal-binding protein homologous to archaeal rhodopsins

    PubMed Central

    Bieszke, Jennifer A.; Braun, Edward L.; Bean, Laura E.; Kang, Seogchan; Natvig, Donald O.; Borkovich, Katherine A.

    1999-01-01

    Opsins are a class of retinal-binding, seven transmembrane helix proteins that function as light-responsive ion pumps or sensory receptors. Previously, genes encoding opsins had been identified in animals and the Archaea but not in fungi or other eukaryotic microorganisms. Here, we report the identification and mutational analysis of an opsin gene, nop-1, from the eukaryotic filamentous fungus Neurospora crassa. The nop-1 amino acid sequence predicts a protein that shares up to 81.8% amino acid identity with archaeal opsins in the 22 retinal binding pocket residues, including the conserved lysine residue that forms a Schiff base linkage with retinal. Evolutionary analysis revealed relatedness not only between NOP-1 and archaeal opsins but also between NOP-1 and several fungal opsin-related proteins that lack the Schiff base lysine residue. The results provide evidence for a eukaryotic opsin family homologous to the archaeal opsins, providing a plausible link between archaeal and visual opsins. Extensive analysis of Δnop-1 strains did not reveal obvious defects in light-regulated processes under normal laboratory conditions. However, results from Northern analysis support light and conidiation-based regulation of nop-1 gene expression, and NOP-1 protein heterologously expressed in Pichia pastoris is labeled by using all-trans [3H]retinal, suggesting that NOP-1 functions as a rhodopsin in N. crassa photobiology. PMID:10393943

  7. Evolving the Living With a Star Data System Definition

    NASA Astrophysics Data System (ADS)

    Otranto, J. F.; Dijoseph, M.

    2003-12-01

    NASA's Living With a Star (LWS) Program is a space weather-focused and applications-driven research program. The LWS Program is soliciting input from the solar, space physics, space weather, and climate science communities to develop a system that enables access to science data associated with these disciplines, and advances the development of discipline and interdisciplinary findings. The LWS Program will implement a data system that builds upon the existing and planned data capture, processing, and storage components put in place by individual spacecraft missions and also inter-project data management systems, including active and deep archives, and multi-mission data repositories. It is technically feasible for the LWS Program to integrate data from a broad set of resources, assuming they are either publicly accessible or allow access by permission. The LWS Program data system will work in coordination with spacecraft mission data systems and science data repositories, integrating their holdings using a common metadata representation. This common representation relies on a robust metadata definition that provides journalistic and technical data descriptions, plus linkages to supporting data products and tools. The LWS Program intends to become an enabling resource to PIs, interdisciplinary scientists, researchers, and students facilitating both access to a broad collection of science data, as well as the necessary supporting components to understand and make productive use of these data. For the LWS Program to represent science data that are physically distributed across various ground system elements, information will be collected about these distributed data products through a series of LWS Program-created agents. These agents will be customized to interface or interact with each one of these data systems, collect information, and forward any new metadata records to a LWS Program-developed metadata library. A populated LWS metadata library will function as a single point-of-contact that serves the entire science community as a first stop for data availability, whether or not science data are physically stored in an LWS-operated repository. Further, this metadata library will provide the user access to information for understanding these data including descriptions of the associated spacecraft and instrument, data format, calibration and operations issues, links to ancillary and correlative data products, links to processing tools and models associated with these data, and any corresponding findings produced using these data. The LWS may also support an active archive for solar, space physics, space weather, and climate data when these data would otherwise be discarded or archived off-line. This archive could potentially serve also as a data storage backup facility for LWS missions. The plan for the LWS Program metadata library is developed based upon input received from the solar and geospace science communities; the library's architecture is based on existing systems developed for serving science metadata. The LWS Program continues to seek constructive input from the science community, examples of both successes and failures in dealing with science data systems, and insights regarding the obstacles between the current state-of-the-practice and this vision for the LWS Program metadata library.

  8. Targeted Research and Technology Within NASA's Living With a Star Program

    NASA Technical Reports Server (NTRS)

    Hesse, Michael

    2003-01-01

    NASA's Living With a Star (LWS) initiative is a systematic, goal-oriented research program targeting those aspects of the Sun-Earth system that affect society. The Targeted Research and Technology (TR&T) component of LWS provides the theory, modeling, and data analysis necessary to enable an integrated, system-wide picture of Sun-Earth connection science with societal relevance. Recognizing the central and essential role that TR&T would have for the success of the LWS initiative, the LWS Science Architecture Team (SAT) recommended that a Science Definition Team (SDT), with the same status as a flight mission definition team, be formed to design and coordinate a TR&T program having prioritized goals and objectives that focused on practical societal benefits. This report details the SDT recommendations for the TR&T program.

  9. [C II] 158-micrometer Observations of a Sample of Late-type Galaxies from the Virgo Cluster

    NASA Technical Reports Server (NTRS)

    Leech, K. J.; Volk, H. J.; Heinrichsen, I.; Hippelein, H.; Metcalfe, L.; Pierini, D.; Popescu, C. C.; Tuffs, R. J.; Xu, C.

    1998-01-01

    We have observed 19 Virgo cluster spiral galaxies with the Long Wavelength Spectrometer (LWS) onboard ESAs Infrared Space Observatory (ISO) obtaining spectral around the (C II) 157.741-micrometer fine structure line.

  10. C II 158 ??bservations of a Sample of Late-type Galaxies from the Virgo Cluster

    NASA Technical Reports Server (NTRS)

    Leech, K.; Volk, H.; Heinrichsen, I.; Hippelein, H.; Metcalfe, L.; Pierini, D.; Popescu, C.; Tuffs, R.; Xu, C.

    1999-01-01

    We have observed 19 Virgo cluster spiral galaxies with the Long Wavelength Spectrometer (LWS) onboard ESAs Infrared Space Observatory (ISO) obtaining spectra around the [CII] 157.741 ??ine structure line.

  11. TARGETED RESEARCH AND TECHNOLOGY WITHIN NASA'S LIVING WITH A STAR PROGRAM.

    SciTech Connect

    Gosling, J. T.; Antiochos, Spiro; Baker, Kile; Bellaire, Paul; Blake, Bern; Crowley, Geoff; Eddy, Jack; Goodrich, Charles; Gopalswamy, Nat; Hesse, Michael; Hurlburt, Neal; Jackman, Charles; Kozyra, Janet; Labonte, Barry; Lean, Judith; Linker, Jon; Mazur, Joe; Onsager, Terry; Sibeck, David

    2003-07-10

    NASA’s Living With a Star (LWS) initiative is a systematic, goal-oriented research program targeting those aspects of the Sun-Earth system that affect society. The Targeted Research and Technology (TR&T) component of LWS provides the theory, modeling, and data analysis necessary to enable an integrated, system-wide picture of Sun-Earth connection science with societal relevance. Recognizing the central and essential role that TR&T would have for the success of the LWS initiative, the LWS Science Architecture Team (SAT) recommended that a Science Definition Team (SDT), with the same status as a flight mission definition team, be formed to design and coordinate a TR&T program having prioritized goals and objectives that focused on practical societal benefits. This report details the SDT recommendations for the TR&T program.

  12. Targeted Research and Technology Within NASA's Living With a Star Program

    NASA Technical Reports Server (NTRS)

    Antiochos, Spiro; Baker, Kile; Bellaire, Paul; Blake, Bern; Crowley, Geoff; Eddy, Jack; Goodrich, Charles; Gopalswamy, Nat; Gosling, Jack; Hesse, Michael

    2004-01-01

    Targeted Research & Technology (TR&T) NASA's Living With a Star (LWS) initiative is a systematic, goal-oriented research program targeting those aspects of the Sun-Earth system that affect society. The Targeted Research and Technology (TR&T) component of LWS provides the theory, modeling, and data analysis necessary to enable an integrated, system-wide picture of Sun-Earth connection science with societal relevance. Recognizing the central and essential role that TR&T would have for the success of the LWS initiative, the LWS Science Architecture Team (SAT) recommended that a Science Definition Team (SDT), with the same status as a flight mission definition team, be formed to design and coordinate a TR&T program having prioritized goals and objectives that focused on practical societal benefits. This report details the SDT recommendations for the TR&T program.

  13. Identification and characterization of a protostome homologue of peropsin from a jumping spider.

    PubMed

    Nagata, Takashi; Koyanagi, Mitsumasa; Tsukamoto, Hisao; Terakita, Akihisa

    2010-01-01

    Peropsin, a member of the opsin family, has characteristics of two functionally distinct opsin-groups, that is, amino acid residues conserved among opsins for light-sensing and a retinal-photoisomerase-like molecular property. Although such a bilateral feature of peropsin seems to be important for understanding the diversity of the opsin family, previous studies have been limited to higher deuterostome, vertebrate and amphioxus peropsins. Here, we report a protostome peropsin homologue from a jumping spider. We found a spider opsin that shares amino acid homology and conserved amino acid residues with known peropsins. The spider opsin-based pigment heterologously expressed in cultured cells exhibited photoisomerase-like isomerization characteristics and a bistable nature. Based on the characteristics of both the amino acid homology and its photochemical properties, we concluded that the spider opsin is the first protostome peropsin homologue. These results show that peropsin existed before the deuterostome-protostome split like other members of the opsin family. In addition, the spider peropsin was localized to non-visual cells in the retina, and fluorescence from reduced retinal chromophore was also observed in the region where peropsin was localized. These findings provide the first demonstration that the peropsin can form a photosensitive pigment in vivo and underlie non-visual function. PMID:19960196

  14. Pathophysilogical Mechanism and Treatment Strategies for Leber Congenital Amaurosis

    PubMed Central

    Zhang, Tao

    2014-01-01

    Mutations in retinoid isomerase, RPE65, or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal recycling and cause Leber congenital amaurosis (LCA), the most severe retinal dystrophy in early childhood. We used Lrat−/−, a murine model for LCA, to investigate the mechanism of rapid cone degeneration. We found that mislocalized M-opsin was degraded whereas mislocalized S-opsin accumulated in Lrat−/− cones before the onset of massive ventral/central cone degeneration. Since the ventral and central retina expresses higher levels of S-opsin than the dorsal retina in mice, our results may explain why ventral and central cones degenerate more rapidly than dorsal cones in Rpe65−/− and Lrat−/− LCA models. In addition, human blue opsin and mouse S-opsin, but not mouse M-opsin or human red/green opsins, aggregated to form cytoplasmic inclusions in transfected cells, which may explain why blue cone function is lost earlier than red/green-cone function in LCA patients. The aggregation of short-wavelength opsins likely caused rapid cone degenerations through an ER stress pathway as demonstrated in both the Lrat−/− retina and transfected cells. Based on this mechanism, we designed a new therapy of LCA by reducing ER stress. We found that systemic injection of an ER chemical chaperone, tauroursodeoxycholic acid (TUDCA), is effective in reducing ER stress, preventing apoptosis, and preserving cones in Lrat−/− mice. PMID:24664772

  15. Pathophysilogical mechanism and treatment strategies for Leber congenital amaurosis.

    PubMed

    Fu, Yingbin; Zhang, Tao

    2014-01-01

    Mutations in retinoid isomerase, RPE65, or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal recycling and cause Leber congenital amaurosis (LCA), the most severe retinal dystrophy in early childhood. We used Lrat (-/-), a murine model for LCA, to investigate the mechanism of rapid cone degeneration. We found that mislocalized M-opsin was degraded whereas mislocalized S-opsin accumulated in Lrat (-/-) cones before the onset of massive ventral/central cone degeneration. Since the ventral and central retina expresses higher levels of S-opsin than the dorsal retina in mice, our results may explain why ventral and central cones degenerate more rapidly than dorsal cones in Rpe65 (-/-) and Lrat (-/-) LCA models. In addition, human blue opsin and mouse S-opsin, but not mouse M-opsin or human red/green opsins, aggregated to form cytoplasmic inclusions in transfected cells, which may explain why blue cone function is lost earlier than red/green-cone function in LCA patients. The aggregation of short-wavelength opsins likely caused rapid cone degenerations through an ER stress pathway as demonstrated in both the Lrat (-/-) retina and transfected cells. Based on this mechanism, we designed a new therapy of LCA by reducing ER stress. We found that systemic injection of an ER chemical chaperone, tauroursodeoxycholic acid (TUDCA), is effective in reducing ER stress, preventing apoptosis, and preserving cones in Lrat (-/-) mice. PMID:24664772

  16. Into the blue: gene duplication and loss underlie color vision adaptations in a deep-sea chimaera, the elephant shark Callorhinchus milii.

    PubMed

    Davies, Wayne L; Carvalho, Livia S; Tay, Boon-Hui; Brenner, Sydney; Hunt, David M; Venkatesh, Byrappa

    2009-03-01

    The cartilaginous fishes reside at the base of the gnathostome lineage as the oldest extant group of jawed vertebrates. Recently, the genome of the elephant shark, Callorhinchus milii, a chimaerid holocephalan, has been sequenced and therefore becomes the first cartilaginous fish to be analyzed in this way. The chimaeras have been largely neglected and very little is known about the visual systems of these fishes. By searching the elephant shark genome, we have identified gene fragments encoding a rod visual pigment, Rh1, and three cone visual pigments, the middle wavelength-sensitive or Rh2 pigment, and two isoforms of the long wavelength-sensitive or LWS pigment, LWS1 and LWS2, but no evidence for the two short wavelength-sensitive cone classes, SWS1 and SWS2. Expression of these genes in the retina was confirmed by RT-PCR. Full-length coding sequences were used for in vitro expression and gave the following peak absorbances: Rh1 496 nm, Rh2 442 nm, LWS1 499 nm, and LWS2 548 nm. Unusually, therefore, for a deep-sea fish, the elephant shark possesses cone pigments and the potential for trichromacy. Compared with other vertebrates, the elephant shark Rh2 and LWS1 pigments are the shortest wavelength-shifted pigments of their respective classes known to date. The mechanisms for this are discussed and we provide experimental evidence that the elephant shark LWS1 pigment uses a novel tuning mechanism to achieve the short wavelength shift to 499 nm, which inactivates the chloride-binding site. Our findings have important implications for the present knowledge of color vision evolution in early vertebrates. PMID:19196633

  17. Dichromatic vision in a fruit bat with diurnal proclivities: the Samoan flying fox (Pteropus samoensis).

    PubMed

    Melin, Amanda D; Danosi, Christina F; McCracken, Gary F; Dominy, Nathaniel J

    2014-12-01

    A nocturnal bottleneck during mammalian evolution left a majority of species with two cone opsins, or dichromatic color vision. Primate trichromatic vision arose from the duplication and divergence of an X-linked opsin gene, and is long attributed to tandem shifts from nocturnality to diurnality and from insectivory to frugivory. Opsin gene variation and at least one duplication event exist in the order Chiroptera, suggesting that trichromatic vision could evolve under favorable ecological conditions. The natural history of the Samoan flying fox (Pteropus samoensis) meets these conditions--it is a large bat that consumes nectar and fruit and demonstrates strong diurnal proclivities. It also possesses a visual system that is strikingly similar to that of primates. To explore the potential for opsin gene duplication and divergence in this species, we sequenced the opsin genes of 11 individuals (19 X-chromosomes) from three South Pacific islands. Our results indicate the uniform presence of two opsins with predicted peak sensitivities of ca. 360 and 553 nm. This result fails to support a causal link between diurnal frugivory and trichromatic vision, although it remains plausible that the diurnal activities of P. samoensis have insufficient antiquity to favor opsin gene renovation. PMID:25319538

  18. Chlamyrhodopsin represents a new type of sensory photoreceptor.

    PubMed Central

    Deininger, W; Kröger, P; Hegemann, U; Lottspeich, F; Hegemann, P

    1995-01-01

    In order to find optimal light conditions for photosynthetic growth, the green alga Chlamydomonas uses a visual system. An optical device, a rhodopsin photoreceptor and an electrical signal transduction chain that mediates between photoreceptor and flagella comprise this system. Here we present an improved strategy for the preparation of eyespot membranes. These membranes contain a retinal binding protein, which has been proposed to be the apoprotein of the phototaxis receptor. The retinal binding protein, which we named chlamyopsin, was purified and opsin-specific antibodies were raised. Using these antibodies, the opsin was localized in the eyespot region of whole cells during growth and cell division. The opsin cDNA was purified and sequenced. The sequence reveals that chlamyopsin is not a typical seven helix receptor. It shows some homology to invertebrate opsins but not to opsins from halobacteria. It contains many polar and charged residues and might function as a light-gated ion channel complex. It is likely that this lower plant rhodopsin diverged from animal opsins early in opsin evolution. Images PMID:8846778

  19. Derivation of Human Differential Photoreceptor-like Cells from the Iris by Defined Combinations of CRX, RX and NEUROD

    PubMed Central

    Seko, Yuko; Azuma, Noriyuki; Kaneda, Makoto; Nakatani, Kei; Miyagawa, Yoshitaka; Noshiro, Yuuki; Kurokawa, Reiko; Okano, Hideyuki; Umezawa, Akihiro

    2012-01-01

    Examples of direct differentiation by defined transcription factors have been provided for beta-cells, cardiomyocytes and neurons. In the human visual system, there are four kinds of photoreceptors in the retina. Neural retina and iris-pigmented epithelium (IPE) share a common developmental origin, leading us to test whether human iris cells could differentiate to retinal neurons. We here define the transcription factor combinations that can determine human photoreceptor cell fate. Expression of rhodopsin, blue opsin and green/red opsin in induced photoreceptor cells were dependent on combinations of transcription factors: A combination of CRX and NEUROD induced rhodopsin and blue opsin, but did not induce green opsin; a combination of CRX and RX induced blue opsin and green/red opsin, but did not induce rhodopsin. Phototransduction-related genes as well as opsin genes were up-regulated in those cells. Functional analysis; i.e. patch clamp recordings, clearly revealed that generated photoreceptor cells, induced by CRX, RX and NEUROD, responded to light. The response was an inward current instead of the typical outward current. These data suggest that photosensitive photoreceptor cells can be generated by combinations of transcription factors. The combination of CRX and RX generate immature photoreceptors: and additional NEUROD promotes maturation. These findings contribute substantially to a major advance toward eventual cell-based therapy for retinal degenerative diseases. PMID:22558175

  20. Insulin-induced hypoglycemia associations with gene expression changes in liver and hypothalamus of chickens from lines selected for low or high body weight.

    PubMed

    Rice, Brittany B; Zhang, Wei; Bai, Shiping; Siegel, Paul B; Cline, Mark A; Gilbert, Elizabeth R

    2014-11-01

    Chickens selected for low (LWS) or high (HWS) body weight for more than 56 generations now have a 10-fold difference in body weight at 56 days of age and correlated responses in appetite and glucose regulation. The LWS chickens are lean and some are anorexic, while the HWS are compulsive feeders and have a different threshold sensitivity of food intake and blood glucose to both central and peripheral insulin, respectively. We previously demonstrated that at 90-days of age, insulin-induced hypoglycemia was associated with reduced glucose transporter expression in the liver of both lines, and differences in expression of neuropeptide Y (NPY) and NPY receptor sub-type genes between LWS and HWS in the hypothalamus. The objective of this study was to determine effects of insulin-induced hypoglycemia on gene expression in the hypothalamus and liver of early post-hatch LWS and HWS chicks. On day 5 post-hatch chicks from each line were fasted for 3h and injected intraperitoneally with insulin or vehicle. At 1h post-injection, chicks were euthanized, blood glucose was measured, and hypothalamus and liver were removed. Total RNA was isolated and real time PCR performed. Insulin injection was associated with a more pronounced reduction in blood glucose in HWS compared with LWS chicks (two-way interaction; P<0.05). Aromatic L-amino acid decarboxylase, NPY, and NPY receptor sub-types 2 and 5 mRNA quantities were greater in LWS than HWS chicks in the hypothalamus (P<0.05), whereas pro-opiomelanocortin mRNA was greater in the hypothalamus of HWS than LWS (P<0.05). In the liver, glucose transporter 1, 2 and 3 (GLUT 1, 2 and 3, respectively) mRNA abundance was greater in HWS than LWS chicks (P<0.05). Compared to the vehicle, insulin treatment was associated with an increase in tryptophan hydroxylase 2 mRNA in the hypothalamus of both lines (P=0.02). In the liver of both lines, insulin treatment was associated with decreased (P=0.01) GLUT2 mRNA and increased (P=0.01) GLUT1 mRNA, compared to vehicle-treated chicks. Results suggest that NPY-associated factors and glucose transporters are differentially-expressed between LWS and HWS chickens and that HWS chicks display greater sensitivity to exogenous insulin during the early post-hatch period. PMID:25157791

  1. Molecular evolution of bat color vision genes.

    PubMed

    Wang, Daryi; Oakley, Todd; Mower, Jeffrey; Shimmin, Lawrence C; Yim, Sokchea; Honeycutt, Rodney L; Tsao, Hsienshao; Li, Wen-Hsiung

    2004-02-01

    The two suborders of bats, Megachiroptera (megabats) and Microchiroptera (microbats), use different sensory modalities for perceiving their environment. Megabats are crepuscular and rely on a well-developed eyes and visual pathway, whereas microbats occupy a nocturnal niche and use acoustic orientation or echolocation more than vision as the major means of perceiving their environment. In view of the differences associated with their sensory systems, we decided to investigate the function and evolution of color vision (opsin genes) in these two suborders of bats. The middle/long wavelength (M/L) and short wavelength (S) opsin genes were sequenced from two frugivorous species of megabats, Haplonycteris fischeri and Pteropus dasymallus formosus, and one insectivorous species of microbat, Myotis velifer. Contrary to the situation in primates, where many nocturnal species have lost the functional S opsin gene, both crepuscular and strictly nocturnal species of bats that we examined have functional M/L and S opsin genes. Surprisingly, the S opsin in these bats may be sensitive to UV light, which is relatively more abundant at dawn and at dusk. The M/L opsin in these bats appears to be the L type, which is sensitive to red and may be helpful for identifying fruits among leaves or for other purposes. Most interestingly, H. fischeri has a recent duplication of the M/L opsin gene, representing to date the only known case of opsin gene duplication in non-primate mammals. Some of these observations are unexpected and may provide insights into the effect of nocturnal life on the evolution of opsin genes in mammals and the evolution of the life history traits of bats in general. PMID:14660703

  2. Strategic Science to Address Current and Future Space Weather Needs

    NASA Astrophysics Data System (ADS)

    Mannucci, A. J.; Schwadron, N.; Antiochos, S. K.; Bhattacharjee, A.; Bisi, M. M.; Gopalswamy, N.; Kamalabadi, F.; Pulkkinen, A. A.; Tobiska, W. K.; Weimer, D. R.; Withers, P.

    2014-12-01

    NASA's Living With a Star (LWS) program has contributed a wealth of scientific knowledge that is relevant to space weather and user needs. A targeted approach to science questions has resulted in leveraging new scientific knowledge to improve not only our understanding of the Heliophysics domain, but also to develop predictive capabilities in key areas of LWS science. This fascinating interplay between science and applications promises to benefit both domains. Scientists providing feedback to the LWS program are now discussing an evolution of the targeted approach that explicitly considers how new science improves, or enables, predictive capability directly. Long-term program goals are termed "Strategic Science Areas" (SSAs) that address predictive capabilities in six specific areas: geomagnetically induced currents, satellite drag, solar energetic particles, ionospheric total electron content, radio frequency scintillation induced by the ionosphere, and the radiation environment. SSAs are organized around user needs and the impacts of space weather on society. Scientists involved in the LWS program identify targeted areas of research that reference (or bear upon) societal needs. Such targeted science leads to new discoveries and is one of the valid forms of exploration. In this talk we describe the benefits of targeted science, and how addressing societal impacts in an appropriate way maintains the strong science focus of LWS, while also leading to its broader impacts.

  3. Role of secondary long wavelength structures in the saturation of electron temperature gradient driven turbulence

    SciTech Connect

    Li Jiquan; Kishimoto, Y.

    2008-11-15

    The dynamics of secondary long wavelength structures (LWSs) in electron temperature gradient (ETG) driven turbulence are investigated by performing gyrofluid simulations and modeling analyses in a slab geometry with an emphasis of the underlying nonlinear interaction processes. It is shown that the back-reaction of the secondary LWS on the ambient fluctuations essentially contributes to saturating ETG instability and limiting the electron transport. The LWS is nonlinearly generated mainly through the beating of the most unstable ETG modes, even a weak modulation instability. The back-reaction is identified as the enhanced stabilization of the ETG modes due to the streamer-type feature of the LWS, which dominantly produces a local poloidal mode coupling among unstable and highly damped spectral components to form a global mode, besides the suppression effect of the LWS due to the radial shearing decorrelation and/or the radial mode coupling. Finally, the correspondence between the LWS in the slab model and the quasimode observed in toroidal ETG simulation [Z. Lin et al., Phys. Plasmas 12, 056125 (2005)] and the importance of the nonlinear mode coupling in the multiscale turbulence interaction are discussed.

  4. Living With a Star, the Geospace Mission Definition Team and Aeronomy

    NASA Technical Reports Server (NTRS)

    Kintner, Paul M., Jr.; Meier, R. R.; Spann, Jim; Whitaker, Ann F. (Technical Monitor)

    2001-01-01

    To gain an understanding of the Sun-Earth system, including how and why the sun varies, how the earth responds, and the impacts on humanity, research is needed that has a integrated and systematic approach. The Living With a Star (LWS) program represents an important element in this regard both to continued progress in space science in general and in Aeronomy in particular. A fundamental question in Aeronomy is how the variable sun affects the ionosphere, thermosphere, and mesosphere. The LWS program focuses on those areas of scientific understanding that promote progress in areas that have human impact and can be investigated with space borne instruments. The Geospace Mission Definition Team is charged with investigating the science priorities identified by the LWS Science Architecture Team and developing an approach to making the necessary measurements in concert with other missions and programs. An important aspect of this approach is that all LWS measurement programs are operating simultaneously for several years. We will review some of the areas that the LWS SAT have emphasized in Aeronomy, including understanding the effects of solar variability on ionospheric density and irregularities, the effects of solar variability on the mass density of the atmosphere at LEO altitudes, and the effects of solar variability on near-surface temperatures and on ozone distribution.

  5. Number and Distribution of Mouse Retinal Cone Photoreceptors: Differences between an Albino (Swiss) and a Pigmented (C57/BL6) Strain

    PubMed Central

    Jiménez-López, Manuel; Alburquerque-Béjar, Juan J.; Nieto-López, Leticia; García-Ayuso, Diego; Villegas-Pérez, Maria P.; Vidal-Sanz, Manuel; Agudo-Barriuso, Marta

    2014-01-01

    We purpose here to analyze and compare the population and topography of cone photoreceptors in two mouse strains using automated routines, and to design a method of retinal sampling for their accurate manual quantification. In whole-mounted retinas from pigmented C57/BL6 and albino Swiss mice, the longwave-sensitive (L) and the shortwave-sensitive (S) opsins were immunodetected to analyze the population of each cone type. In another group of retinas both opsins were detected with the same fluorophore to quantify all cones. In a third set of retinas, L-opsin and Brn3a were immunodetected to determine whether L-opsin+cones and retinal ganglion cells (RGCs) have a parallel distribution. Cones and RGCs were automatically quantified and their topography illustrated with isodensity maps. Our results show that pigmented mice have a significantly higher number of total cones (all-cones) and of L-opsin+cones than albinos which, in turn, have a higher population of S-opsin+cones. In pigmented animals 40% of cones are dual (cones that express both opsins), 34% genuine-L (cones that only express the L-opsin), and 26% genuine-S (cones that only express the S-opsin). In albinos, 23% of cones are genuine-S and the proportion of dual cones increases to 76% at the expense of genuine-L cones. In both strains, L-opsin+cones are denser in the central than peripheral retina, and all-cones density increases dorso-ventrally. In pigmented animals S-opsin+cones are scarce in the dorsal retina and very numerous in the ventral retina, being densest in its nasal aspect. In albinos, S-opsin+cones are abundant in the dorsal retina, although their highest densities are also ventral. Based on the densities of each cone population, we propose a sampling method to manually quantify and infer their total population. In conclusion, these data provide the basis to study cone degeneration and its prevention in pathologic conditions. PMID:25029531

  6. Total Land Water Storage Change over 2003 - 2013 Estimated from a Global Mass Budget Approach

    NASA Technical Reports Server (NTRS)

    Dieng, H. B.; Champollion, N.; Cazenave, A.; Wada, Y.; Schrama, E.; Meyssignac, B.

    2015-01-01

    We estimate the total land water storage (LWS) change between 2003 and 2013 using a global water mass budget approach. Hereby we compare the ocean mass change (estimated from GRACE space gravimetry on the one hand, and from the satellite altimetry-based global mean sea level corrected for steric effects on the other hand) to the sum of the main water mass components of the climate system: glaciers, Greenland and Antarctica ice sheets, atmospheric water and LWS (the latter being the unknown quantity to be estimated). For glaciers and ice sheets, we use published estimates of ice mass trends based on various types of observations covering different time spans between 2003 and 2013. From the mass budget equation, we derive a net LWS trend over the study period. The mean trend amounts to +0.30 +/- 0.18 mm/yr in sea level equivalent. This corresponds to a net decrease of -108 +/- 64 cu km/yr in LWS over the 2003-2013 decade. We also estimate the rate of change in LWS and find no significant acceleration over the study period. The computed mean global LWS trend over the study period is shown to be explained mainly by direct anthropogenic effects on land hydrology, i.e. the net effect of groundwater depletion and impoundment of water in man-made reservoirs, and to a lesser extent the effect of naturally-forced land hydrology variability. Our results compare well with independent estimates of human-induced changes in global land hydrology.

  7. Total land water storage change over 2003-2013 estimated from a global mass budget approach

    NASA Astrophysics Data System (ADS)

    Dieng, H. B.; Champollion, N.; Cazenave, A.; Wada, Y.; Schrama, E.; Meyssignac, B.

    2015-12-01

    We estimate the total land water storage (LWS) change between 2003 and 2013 using a global water mass budget approach. Hereby we compare the ocean mass change (estimated from GRACE space gravimetry on the one hand, and from the satellite altimetry-based global mean sea level corrected for steric effects on the other hand) to the sum of the main water mass components of the climate system: glaciers, Greenland and Antarctica ice sheets, atmospheric water and LWS (the latter being the unknown quantity to be estimated). For glaciers and ice sheets, we use published estimates of ice mass trends based on various types of observations covering different time spans between 2003 and 2013. From the mass budget equation, we derive a net LWS trend over the study period. The mean trend amounts to +0.30 ± 0.18 mm yr-1 in sea level equivalent. This corresponds to a net decrease of -108 ± 64 km3 yr-1 in LWS over the 2003-2013 decade. We also estimate the rate of change in LWS and find no significant acceleration over the study period. The computed mean global LWS trend over the study period is shown to be explained mainly by direct anthropogenic effects on land hydrology, i.e. the net effect of groundwater depletion and impoundment of water in man-made reservoirs, and to a lesser extent the effect of naturally-forced land hydrology variability. Our results compare well with independent estimates of human-induced changes in global land hydrology.

  8. Breaking the Covalent Bond—A Pigment Property that Contributes to Desensitization in Cones

    PubMed Central

    Kefalov, Vladimir J.; Estevez, Maureen E.; Kono, Massahiro; Goletz, Patrice W.; Crouch, Rosalie K.; Cornwall, M. Carter; Yau, King-Wai

    2010-01-01

    Summary Retinal rod and cone pigments consist of an apoprotein, opsin, covalently linked to a chromophore, 11-cis retinal. Here we demonstrate that the formation of the covalent bond between opsin and 11-cis retinal is reversible in darkness in amphibian red cones, but essentially irreversible in red rods. This dissociation, apparently a general property of cone pigments, results in a surprisingly large amount of free opsin—about 10% of total opsin—in dark-adapted red cones. We attribute this significant level of free opsin to the low concentration of intracellular free 11-cis retinal, estimated to be only a tiny fraction (~0.1 %) of the pigment content in red cones. With its constitutive transducin-stimulating activity, the free cone opsin produces an ~2-fold desensitization in red cones, equivalent to that produced by a steady light causing 500 photoisomerizations s−1. Cone pigment dissociation therefore contributes to the sensitivity difference between rods and cones. PMID:15953417

  9. The molecular basis for the green-blue sensitivity shift in the rod visual pigments of the European eel.

    PubMed

    Archer, S; Hope, A; Partridge, J C

    1995-12-22

    When the European eel matures sexually and migrates back to deep sea breeding grounds the visual pigments in its rod photoreceptors change from being maximally sensitive to green light to being maximally sensitive to blue light. In part, this change in sensitivity is due to a change in the opsin component of the visual pigment molecule. We used hormone injection to induce these developmental changes in a group of eels and from these animals an opsin coding region was cloned and sequenced using cDNA made from retinal mRNA. From the retinae of hormone-injected eels and those not injected with hormones, distinct opsin mRNAs were isolated. These mRNAs encode two rod opsin proteins that are very similar but have significant amino acid substitutions in key positions that are likely to be involved in spectral tuning of the eel green and blue sensitive rod visual pigment molecules. PMID:8587887

  10. The photochemical determinants of color vision

    PubMed Central

    Wang, Wenjing; Geiger, James H; Borhan, Babak

    2014-01-01

    The evolution of a variety of important chromophore-dependent biological processes, including microbial light sensing and mammalian color vision, relies on protein modifications that alter the spectral characteristics of a bound chromophore. Three different color opsins share the same chromophore, but have three distinct absorptions that together cover the entire visible spectrum, giving rise to trichromatic vision. The influence of opsins on the absorbance of the chromophore has been studied through methods such as model compounds, opsin mutagenesis, and computational modeling. The recent development of rhodopsin mimic that uses small soluble proteins to recapitulate the binding and wavelength tuning of the native opsins provides a new platform for studying protein-regulated spectral tuning. The ability to achieve far-red shifted absorption in the rhodopsin mimic system was attributed to a combination of the lack of a counteranion proximal to the iminium, and a uniformly neutral electrostatic environment surrounding the chromophore. PMID:24323922

  11. The Living With a Star Program Space Environment Testbed

    NASA Technical Reports Server (NTRS)

    Barth, Janet; Day, John H. (Technical Monitor)

    2001-01-01

    This viewgraph presentation describes the objective, approach, and scope of the Living With a Star (LWS) program at the Marshall Space Flight Center. Scientists involved in the project seek to refine the understanding of space weather and the role of solar variability in terrestrial climate change. Research and the development of improved analytic methods have led to increased predictive capabilities and the improvement of environment specification models. Specifically, the Space Environment Testbed (SET) project of LWS is responsible for the implementation of improved engineering approaches to observing solar effects on climate change. This responsibility includes technology development, ground test protocol development, and the development of a technology application model/engineering tool.

  12. Living with a Star Space Environment Testbed

    NASA Technical Reports Server (NTRS)

    Barth, Janet

    2003-01-01

    Summary of activities: (1) FYO1 NRA - Model development and data mining. (2) FY03 NRA - Flight investigations. (3) SET carrier development. (4) Study for accommodation of SET carrier to support advanced detectors. (5) Collaboration with other programs: LWS TR&T to maximize synergy between TR&T space environment research and SET space environment effects research. LWS Data System to optimize dissemination of SET data. NASA Electronic Parts and Packaging Program to leverage ground testing of technologies. Defense Threat Reduction Agency to leverage ground testing and common interests in advanced detectors. and Air Force Research Laboratory to leverage flight opportunities. (6) Education and Public Outreach.

  13. Seismic properties of subducting oceanic crust: Constraints from natural lawsonite-bearing blueschist and eclogite in Sivrihisar Massif, Turkey

    NASA Astrophysics Data System (ADS)

    Cao, Yi; Jung, Haemyeong

    2016-01-01

    Investigating the seismic properties of natural lawsonite (Lws)-bearing blueschist and eclogite is particularly important for constraining the seismic interpretation of subducting oceanic crust based on seismological observations. To achieve this end, we analyzed in detail the mineral fabrics and seismic properties of foliated Lws-blueschist and Lws-eclogites from Sivrihisar Massif in Turkey. In both blueschists and eclogites, the lawsonite fabric is characterized by three different patterns: [0 0 1] axes aligning sub-normal to foliation, and [0 1 0] axes aligning sub-parallel to lineation (normal type); [0 0 1] axes aligning sub-parallel to lineation, and [1 0 0] axes aligning sub-normal to foliation with a girdle sub-normal to lineation (abnormal type); and [0 0 1] axes aligning both sub-normal to foliation and sub-parallel to lineation, [0 1 0] axes aligning sub-parallel to lineation, and [1 0 0] axes aligning sub-normal to foliation (transitional pattern). In contrast, glaucophane and omphacite mostly present consistent axial fabrics with the [0 0 1] axes aligning to lineation. These mineral fabrics produce whole-rock seismic anisotropies with similar patterns. However, the variations in seismic anisotropies are mainly controlled by the rock type, to a lesser extent are determined by the lawsonite fabric type, and to only a small extent are affected by mineral fabric strength. Despite the constructive abnormal-type lawsonite fabric on whole-rock seismic anisotropies, because of their weaker mineral fabric strength (or deformation degree), the abnormal-type Lws-blueschist still exhibit comparatively lower seismic anisotropies than those normal-type Lws-blueschist from other localities. Based on the calculated seismic anisotropies and velocities, we estimated that when oceanic crust transforms from Lws-blueschist to Lws-eclogite with increasing subduction depth, (1) P-wave and max. S-wave polarization anisotropies reduce about 70% and 40%, respectively; and (2) variations of Vp and Vs contrasts relative to mantle peridotites are about -7% to -3% and -8% to -6%, respectively. These results corroborate the important roles of Lws-bearing blueschist and eclogite in interpreting the existence and gradual weakening of low-velocity layers in subducting oceanic crust, during the subduction process.

  14. Activation of Transducin by Bistable Pigment Parapinopsin in the Pineal Organ of Lower Vertebrates.

    PubMed

    Kawano-Yamashita, Emi; Koyanagi, Mitsumasa; Wada, Seiji; Tsukamoto, Hisao; Nagata, Takashi; Terakita, Akihisa

    2015-01-01

    Pineal organs of lower vertebrates contain several kinds of photosensitive molecules, opsins that are suggested to be involved in different light-regulated physiological functions. We previously reported that parapinopsin is an ultraviolet (UV)-sensitive opsin that underlies hyperpolarization of the pineal photoreceptor cells of lower vertebrates to achieve pineal wavelength discrimination. Although, parapinopsin is phylogenetically close to vertebrate visual opsins, it exhibits a property similar to invertebrate visual opsins and melanopsin: the photoproduct of parapinopsin is stable and reverts to the original dark states, demonstrating the nature of bistable pigments. Therefore, it is of evolutionary interest to identify a phototransduction cascade driven by parapinopsin and to compare it with that in vertebrate visual cells. Here, we showed that parapinopsin is coupled to vertebrate visual G protein transducin in the pufferfish, zebrafish, and lamprey pineal organs. Biochemical analyses demonstrated that parapinopsins activated transducin in vitro in a light-dependent manner, similar to vertebrate visual opsins. Interestingly, transducin activation by parapinopsin was provoked and terminated by UV- and subsequent orange-lights irradiations, respectively, due to the bistable nature of parapinopsin, which could contribute to a wavelength-dependent control of a second messenger level in the cell as a unique optogenetic tool. Immunohistochemical examination revealed that parapinopsin was colocalized with Gt2 in the teleost, which possesses rod and cone types of transducin, Gt1, and Gt2. On the other hand, in the lamprey, which does not possess the Gt2 gene, in situ hybridization suggested that parapinopsin-expressing photoreceptor cells contained Gt1 type transducin GtS, indicating that lamprey parapinopsin may use GtS in place of Gt2. Because it is widely accepted that vertebrate visual opsins having a bleaching nature have evolved from non-bleaching opsins similar to parapinopsin, these results implied that ancestral bistable opsins might acquire coupling to the transducin-mediated cascade and achieve light-dependent hyperpolarizing response of the photoreceptor cells. PMID:26492337

  15. Activation of Transducin by Bistable Pigment Parapinopsin in the Pineal Organ of Lower Vertebrates

    PubMed Central

    Kawano-Yamashita, Emi; Koyanagi, Mitsumasa; Wada, Seiji; Tsukamoto, Hisao; Nagata, Takashi; Terakita, Akihisa

    2015-01-01

    Pineal organs of lower vertebrates contain several kinds of photosensitive molecules, opsins that are suggested to be involved in different light-regulated physiological functions. We previously reported that parapinopsin is an ultraviolet (UV)-sensitive opsin that underlies hyperpolarization of the pineal photoreceptor cells of lower vertebrates to achieve pineal wavelength discrimination. Although, parapinopsin is phylogenetically close to vertebrate visual opsins, it exhibits a property similar to invertebrate visual opsins and melanopsin: the photoproduct of parapinopsin is stable and reverts to the original dark states, demonstrating the nature of bistable pigments. Therefore, it is of evolutionary interest to identify a phototransduction cascade driven by parapinopsin and to compare it with that in vertebrate visual cells. Here, we showed that parapinopsin is coupled to vertebrate visual G protein transducin in the pufferfish, zebrafish, and lamprey pineal organs. Biochemical analyses demonstrated that parapinopsins activated transducin in vitro in a light-dependent manner, similar to vertebrate visual opsins. Interestingly, transducin activation by parapinopsin was provoked and terminated by UV- and subsequent orange-lights irradiations, respectively, due to the bistable nature of parapinopsin, which could contribute to a wavelength-dependent control of a second messenger level in the cell as a unique optogenetic tool. Immunohistochemical examination revealed that parapinopsin was colocalized with Gt2 in the teleost, which possesses rod and cone types of transducin, Gt1, and Gt2. On the other hand, in the lamprey, which does not possess the Gt2 gene, in situ hybridization suggested that parapinopsin-expressing photoreceptor cells contained Gt1 type transducin GtS, indicating that lamprey parapinopsin may use GtS in place of Gt2. Because it is widely accepted that vertebrate visual opsins having a bleaching nature have evolved from non-bleaching opsins similar to parapinopsin, these results implied that ancestral bistable opsins might acquire coupling to the transducin-mediated cascade and achieve light-dependent hyperpolarizing response of the photoreceptor cells. PMID:26492337

  16. Encephalic photoreception and phototactic response in the troglobiont Somalian blind cavefish Phreatichthys andruzzii.

    PubMed

    Tarttelin, Emma E; Frigato, Elena; Bellingham, James; Di Rosa, Viviana; Berti, Roberto; Foulkes, Nicholas S; Lucas, Robert J; Bertolucci, Cristiano

    2012-08-15

    Many physiological and behavioural responses to changes in environmental lighting conditions are mediated by extraocular photoreceptors. Here we investigate encephalic photoreception in Phreatichthys andruzzii, a typical cave-dwelling fish showing an extreme phenotype with complete anophthalmy and a reduction in size of associated brain structures. We firstly identified two P. andruzzii photopigments, orthologues of rod opsin and exo-rod opsin. In vitro, both opsins serve as light-absorbing photopigments with λ(max) around 500 nm when reconstituted with an A(1) chromophore. When corrected for the summed absorption from the skin and skull, the spectral sensitivity profiles shifted to longer wavelengths (rod opsin: 521 nm; exo-rod opsin: 520 nm). We next explored the involvement of both opsins in the negative phototaxis reported for this species. A comparison of the spectral sensitivity of the photophobic response with the putative A(2) absorbance spectra corrected for skin/skull absorbance indicates that the A(2) versions of either or both of these pigments could explain the observed behavioural spectral sensitivity. PMID:22837464

  17. An extended family of novel vertebrate photopigments is widely expressed and displays a diversity of function.

    PubMed

    Davies, Wayne I L; Tamai, T Katherine; Zheng, Lei; Fu, Josephine K; Rihel, Jason; Foster, Russell G; Whitmore, David; Hankins, Mark W

    2015-11-01

    Light affects animal physiology and behavior more than simply through classical visual, image-forming pathways. Nonvisual photoreception regulates numerous biological systems, including circadian entrainment, DNA repair, metabolism, and behavior. However, for the majority of these processes, the photoreceptive molecules involved are unknown. Given the diversity of photophysiological responses, the question arises whether a single photopigment or a greater diversity of proteins within the opsin superfamily detect photic stimuli. Here, a functional genomics approach identified the full complement of photopigments in a highly light-sensitive model vertebrate, the zebrafish (Danio rerio), and characterized their tissue distribution, expression levels, and biochemical properties. The results presented here reveal the presence of 42 distinct genes encoding 10 classical visual photopigments and 32 nonvisual opsins, including 10 novel opsin genes comprising four new pigment classes. Consistent with the presence of light-entrainable circadian oscillators in zebrafish, all adult tissues examined expressed two or more opsins, including several novel opsins. Spectral and electrophysiological analyses of the new opsins demonstrate that they form functional photopigments, each with unique chromophore-binding and wavelength specificities. This study has revealed a remarkable number and diversity of photopigments in zebrafish, the largest number so far discovered for any vertebrate. Found in amphibians, reptiles, birds, and all three mammalian clades, most of these genes are not restricted to teleosts. Therefore, nonvisual light detection is far more complex than initially appreciated, which has significant biological implications in understanding photoreception in vertebrates. PMID:26450929

  18. A butterfly eye's view of birds.

    PubMed

    Frentiu, Francesca D; Briscoe, Adriana D

    2008-11-01

    The striking color patterns of butterflies and birds have long interested biologists. But how these animals see color is less well understood. Opsins are the protein components of the visual pigments of the eye. Color vision has evolved in butterflies through opsin gene duplications, through positive selection at individual opsin loci, and by the use of filtering pigments. By contrast, birds have retained the same opsin complement present in early-jawed vertebrates, and their visual system has diversified primarily through tuning of the short-wavelength-sensitive photoreceptors, rather than by opsin duplication or the use of filtering elements. Butterflies and birds have evolved photoreceptors that might use some of the same amino acid sites for generating similar spectral phenotypes across approximately 540 million years of evolution, when rhabdomeric and ciliary-type opsins radiated during the early Cambrian period. Considering the similarities between the two taxa, it is surprising that the eyes of birds are not more diverse. Additional taxonomic sampling of birds may help clarify this mystery. PMID:18937365

  19. Immunohistochemical evidence for multiple photosystems in box jellyfish.

    PubMed

    Ekström, Peter; Garm, Anders; Pålsson, Jonas; Vihtelic, Thomas S; Nilsson, Dan-Eric

    2008-07-01

    Cubomedusae (box jellyfish) possess a remarkable visual system with 24 eyes distributed in four sensory structures termed rhopalia. Each rhopalium is equipped with six eyes: two pairs of pigment cup eyes and two unpaired lens eyes. Each eye type probably captures specific features of the visual environment. To investigate whether multiple types of photoreceptor cells are present in the rhopalium, and whether the different eye types possess different types of photoreceptors, we have used immunohistochemistry with a range of vertebrate opsin antibodies to label the photoreceptors, and electroretinograms (ERG) to determine their spectral sensitivity. All photoreceptor cells of the two lens eyes of the box jellyfish Tripedalia cystophora and Carybdea marsupialis displayed immunoreactivity for an antibody directed against the zebrafish ultraviolet (UV) opsin, but not against any of eight other rhodopsin or cone opsin antibodies tested. In neither of the two species were the pigment cup eyes immunoreactive for any of the opsin antibodies. ERG analysis of the Carybdea lower lens eyes demonstrated a single spectral sensitivity maximum at 485 nm suggesting the presence of a single opsin type. Our data demonstrate that the lens eyes of box jellyfish utilize a single opsin and are thus color-blind, and that there is probably a different photopigment in the pigment cup eyes. The results support our hypothesis that the lens eyes and the pigment cup eyes of box jellyfish are involved in different and specific visual tasks. PMID:18504619

  20. Human Cone Visual Pigment Deletions Spare Sufficient Photoreceptors to Warrant Gene Therapy

    PubMed Central

    Cideciyan, Artur V.; Hufnagel, Robert B.; Carroll, Joseph; Sumaroka, Alexander; Luo, Xunda; Schwartz, Sharon B.; Dubra, Alfredo; Land, Megan; Michaelides, Michel; Gardner, Jessica C.; Hardcastle, Alison J.; Moore, Anthony T.; Sisk, Robert A.; Ahmed, Zubair M.; Kohl, Susanne

    2013-01-01

    Abstract Human X-linked blue-cone monochromacy (BCM), a disabling congenital visual disorder of cone photoreceptors, is a candidate disease for gene augmentation therapy. BCM is caused by either mutations in the red (OPN1LW) and green (OPN1MW) cone photoreceptor opsin gene array or large deletions encompassing portions of the gene array and upstream regulatory sequences that would predict a lack of red or green opsin expression. The fate of opsin-deficient cone cells is unknown. We know that rod opsin null mutant mice show rapid postnatal death of rod photoreceptors. Using in vivo histology with high-resolution retinal imaging, we studied a cohort of 20 BCM patients (age range 5–58) with large deletions in the red/green opsin gene array. Already in the first years of life, retinal structure was not normal: there was partial loss of photoreceptors across the central retina. Remaining cone cells had detectable outer segments that were abnormally shortened. Adaptive optics imaging confirmed the existence of inner segments at a spatial density greater than that expected for the residual blue cones. The evidence indicates that human cones in patients with deletions in the red/green opsin gene array can survive in reduced numbers with limited outer segment material, suggesting potential value of gene therapy for BCM. PMID:24067079

  1. An extended family of novel vertebrate photopigments is widely expressed and displays a diversity of function

    PubMed Central

    Davies, Wayne I.L.; Tamai, T. Katherine; Zheng, Lei; Fu, Josephine K.; Rihel, Jason; Foster, Russell G.; Whitmore, David; Hankins, Mark W.

    2015-01-01

    Light affects animal physiology and behavior more than simply through classical visual, image-forming pathways. Nonvisual photoreception regulates numerous biological systems, including circadian entrainment, DNA repair, metabolism, and behavior. However, for the majority of these processes, the photoreceptive molecules involved are unknown. Given the diversity of photophysiological responses, the question arises whether a single photopigment or a greater diversity of proteins within the opsin superfamily detect photic stimuli. Here, a functional genomics approach identified the full complement of photopigments in a highly light-sensitive model vertebrate, the zebrafish (Danio rerio), and characterized their tissue distribution, expression levels, and biochemical properties. The results presented here reveal the presence of 42 distinct genes encoding 10 classical visual photopigments and 32 nonvisual opsins, including 10 novel opsin genes comprising four new pigment classes. Consistent with the presence of light-entrainable circadian oscillators in zebrafish, all adult tissues examined expressed two or more opsins, including several novel opsins. Spectral and electrophysiological analyses of the new opsins demonstrate that they form functional photopigments, each with unique chromophore-binding and wavelength specificities. This study has revealed a remarkable number and diversity of photopigments in zebrafish, the largest number so far discovered for any vertebrate. Found in amphibians, reptiles, birds, and all three mammalian clades, most of these genes are not restricted to teleosts. Therefore, nonvisual light detection is far more complex than initially appreciated, which has significant biological implications in understanding photoreception in vertebrates. PMID:26450929

  2. Numerical Analysis of Effects of Large Wood Structures on Channel Morphology and Fish Habitat Suitability in a Southern U.S. Sandy Creek

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A depth-averaged two-dimensional model was applied to simulate the effect of large wood structures (LWS) on flow, sediment transport, bed change, and fish habitat in a deeply-incised sharp bend in the Little Topashaw Creek, North Central Mississippi. The hydrodynamic simulation showed that the flow ...

  3. Numerical analysis of effects of large wood structures on channel morphology and fish habitat suitability in Little Topashaw Creek

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A depth-averaged two dimensional (2D) model has been applied to simulate the effect of large wood structures (LWS) on flow, sediment transport, bed change, and fish habitat suitability in a deeply-incised sharp bend in the Little Topashaw Creek, North Central Mississippi. The hydrodynamic simulation...

  4. The contribution of IFSI (Istituto di Fisica dello Spazio Interplanetario) to the ISO project

    NASA Astrophysics Data System (ADS)

    Orfei, R.; Baldetti, P.; Ceccarelli, C.; Cerulli, P.; Lorenzetti, D.

    1990-02-01

    IFSI efforts in the development of the long-wavelength spectrometer (LWS) for the IR Space Observatory (ISO), scheduled for launch in 1993, are briefly reviewed. The LWS operates at 45-200 microns with resolution of about 200 (using a grating alone) or about 10,000 (using a grating with one of two Fabry-Perot etalons). Other LWS parameters include FOV 1.5 arcmin, sensitivity 1 x 10 to the -16th W/sq m, and SNR = 10 for a 10-sec high-resolution observation. Particular attention is given to the LWS data-processing unit (DPU), based on an 80C86 CPU and providing telecommand reception at 80 bps, data telemetry at 32,000 bps, detector sampling once every 0.5 msec, control of gratings and etalons, and continuous self-maintenance. Since the orbit of ISO exposes it to the earth radiation belts for about 2 h/day, the DPU is doubled, and the software is completely relocatable. An overview of the software design and a description of the low-noise dc-dc converter are included.

  5. FIELD EVALUATION OF LOW-EMISSION COAL BURNER TECHNOLOGY ON UTILITY BURNERS VOLUME V. BURNER EVALUATION DATA APPENDICES

    EPA Science Inventory

    The report gives a detailed summary of data which were generated during the testing of experimental burners on EPA's Large Watertube Simulator (LWS) test facility. The test data are presented as a series of appendices. Appendix A describe the data quality assurance procedures whi...

  6. 75 FR 81218 - Laminated Woven Sacks From the People's Republic of China: Preliminary Results of the Second...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-27

    ...The Department of Commerce (``the Department'') is currently conducting an administrative review of the antidumping duty order on laminated woven sacks (``LWS'') from the People's Republic of China (``PRC'') covering the period August 1, 2009, through July 31, 2010. This review covers imports of subject merchandise from one manufacturer/exporter: Zibo Aifudi Plastic Packaging Co., Ltd.......

  7. Internal P-T-t Structure of Subduction Complexes — Insights from Lu-Hf Geochronology on Garnet and Lawsonite (Halilbağı, Central Anatolia)

    NASA Astrophysics Data System (ADS)

    Pourteau, A.; Scherer, E. E.; Schmidt, A.; Bast, R.

    2014-12-01

    The subduction complex near Halilbağı (Central Anatolia) is among the best sites to investigate deep-seated tectonic, petrologic, and geochemical processes taking place in subduction zones. The Halilbağı Unit comprises slices of lawsonite- and/or epidote-bearing blueschist and eclogite, as well as meta-chert and marble. The unit is overlain by an ophiolitic slab and underlain by a HP/LT metamorphosed carbonate platform. Previous studies of the Halilbağı Unit suggested tectonic blocks were metamorphosed under diverse peak conditions, but shared a common exhumation P-T path marked by syn-decompression cooling (Davis and Whitney, 2006; Çetinkaplan et al., 2008). To better understand the internal structure and dynamics of this subduction complex, we carried out Lu-Hf geochronology on garnet (grt) and lawsonite (lws) from a variety of HP oceanic rocks, as well as the sub-ophiolitic metamorphic sole. Our results suggest that intra-oceanic subduction started at ~110 Ma (grt-amph isochron from a grt amphibolite). Less than 23 Myr later, the subduction interface was refrigerated enough to allow clockwise P-T loops (~87 Ma peak grt-matrix isochron for a lws+grt-bearing eclogitic blueschist) and syn-decompression cooling (~79 Ma retrograde lws-matrix isochron) of subducted oceanic rocks. We will present further results for several HP metamorphic sub-facies (e.g., epidote (ep) eclogite, lws+ep blueschist, lws blueschist, lws eclogite). Such data may allow unraveling whether the co-occurrence of "warm" (i.e., ep-bearing), and "cold" (i.e., lws-bearing) HP rocks in the same locality results from (a) sampling of distinct levels of the subduction slab, (b) thermal maturation of the juvenile subduction zone, or (c) inaccurate P-T estimates. Novel natural constraints are thus expected on the dynamics of the Halilbağı Unit and of subduction complexes in general. Çetinkaplan M., Candan O., Oberhänsli R. and Bousquet R., 2008. Pressure-Temperature Evolution of Lawsonite Eclogite in Sivrihisar, Tavşanlı Zone-Turkey. Lithos, 104(1-4): 12-32. Davis P. B. and Whitney D. L., 2006. Petrogenesis of Lawsonite and Epidote Eclogite and Blueschist, Sivrihisar Massif, Turkey. Journal of Metamorphic Geology, 24(9): 823-849.

  8. Fed and fasted chicks from lines divergently selected for low or high body weight have differential hypothalamic appetite-associated factor mRNA expression profiles.

    PubMed

    Yi, Jiaqing; Gilbert, Elizabeth R; Siegel, Paul B; Cline, Mark A

    2015-06-01

    We have demonstrated that chicken lines which have undergone intense divergent selection for either low (LWS) or high (HWS) body weight (anorexic and obese containing, respectively) have differential food intake threshold responses to a range of intracerebroventricular injected neurotransmitters. The study reported herein was designed to measure endogenous appetite-associated factor mRNA profiles between these lines in an effort to further understand the molecular mechanisms involved in their differential eating patterns. Whole hypothalamus was collected from 5 day-old chicks that had been fasted for 180 min or had free access to food. Total RNA was isolated, reverse transcribed, and real-time PCR performed. Although mRNAs encoding orexigenic neuropeptides including agouti-related peptide, neuropeptide Y (NPY), prolactin-releasing peptide, and visfatin did not differ in expression between the lines, NPY receptor 5 mRNA was greater in fed LWS than HWS chicks, but fasting decreased the magnitude of difference. Anorexigenic factors including amylin, corticotropin releasing factor (CRF) and ghrelin were not differentially expressed between lines, while mRNA abundance of calcitonin, CRF receptor 1, leptin receptor, neuropeptide S, melanocortin receptor 3, and oxytocin were greater in LWS than HWS chicks. Pro-opiomelanocortin mRNA was lower in LWS than HWS chicks, while fasting decreased its expression in both lines. These results suggest that there are differences in gene expression of appetite-associated factors between LWS and HWS lines that might be associated with their differential food intake and thus contribute to differences in severity of anorexia, body weight, adiposity, and development of obesity. PMID:25677648

  9. Photoreceptor topography and spectral sensitivity in the common brushtail possum (Trichosurus vulpecula).

    PubMed

    Vlahos, Lisa M; Knott, Ben; Valter, Krisztina; Hemmi, Jan M

    2014-10-15

    Marsupials are believed to be the only non-primate mammals with both trichromatic and dichromatic color vision. The diversity of color vision systems present in marsupials remains mostly unexplored. Marsupials occupy a diverse range of habitats, which may have led to considerable variation in the presence, density, distribution, and spectral sensitivity of retinal photoreceptors. In this study we analyzed the distribution of photoreceptors in the common brushtail possum (Trichosurus vulpecula). Immunohistochemistry in wholemounts revealed three cone subpopulations recognized within two spectrally distinct cone classes. Long-wavelength sensitive (LWS) single cones were the largest cone subgroup (67-86%), and formed a weak horizontal visual streak (peak density 2,106 ± 435/mm2) across the central retina. LWS double cones were strongly concentrated ventrally (569 ± 66/mm2), and created a "negative" visual streak (134 ± 45/mm2) in the central retina. The strong regionalization between LWS cone topographies suggests differing visual functions. Short-wavelength sensitive (SWS) cones were present in much lower densities (3-10%), mostly located ventrally (179 ± 101/mm2). A minority population of cones (0-2.4%) remained unlabeled by both SWS- and LWS-specific antibodies, and may represent another cone population. Microspectrophotometry of LWS cone and rod visual pigments shows peak spectral sensitivities at 544 nm and 500 nm, respectively. Cone to ganglion cell convergences remain low and constant across the retina, thereby maintaining good visual acuity, but poor contrast sensitivity during photopic vision. Given that brushtail possums are so strongly nocturnal, we hypothesize that their acuity is set by the scotopic visual system, and have minimized the number of cones necessary to serve the ganglion cells for photopic vision. PMID:24737644

  10. A Cluster Randomized Trial to Evaluate a Health Education Programme “Living with Sun at School”

    PubMed Central

    Sancho-Garnier, Hélène; Pereira, Bruno; Césarini, Pierre

    2012-01-01

    Over-exposure to sunlight increases the risk of skin cancers, particularly when exposure occurs during childhood. School teachers can play an active role in providing an education programme that can help prevent this. “Living with the Sun,” (LWS) is a sun safety education program for school children based on a handy guide for classroom activities designed to improve children’s knowledge, but moreover to positively modify their sun safety attitudes and behaviours. The goal of our study was to determine the effectiveness of this programme by examining children’s knowledge, attitude and sun exposure behaviours prior to and after the completion of the programme. We carried out a cluster randomised trial in which the classes were randomly assigned to one of two groups; one using the LWS programme and another that didn’t, serving as the control. Data was collected before completion of the programme and an additional three times in the year after completion. The 70 participating classes (1,365 schoolchildren) were distributed throughout France. Statistical analysis confirmed that knowledge of sun risk increased significantly in the LWS classes (p < 0.001). Both groups positively changed their attitudes when considering the best sun protection, but the LWS group proved to consistently be more convinced (p = 0.04). After the summer holidays, differences between the two groups decreased throughout the year but stayed globally significant. We also observed some significant behaviour modification during the holidays. For instance, the LWS group applied sunscreen more frequently than the control group, and were more likely to wear a hat (72% versus 59%) and use a sun umbrella on the beach (75% versus 64%). PMID:22851947

  11. The evolution of complexity in the visual systems of stomatopods: insights from transcriptomics.

    PubMed

    Porter, Megan L; Speiser, Daniel I; Zaharoff, Alexander K; Caldwell, Roy L; Cronin, Thomas W; Oakley, Todd H

    2013-07-01

    Stomatopod crustaceans have complex visual systems containing up to 16 different spectral classes of photoreceptors, more than described for any other animal. A previous molecular study of this visual system focusing on the expression of opsin genes found many more transcripts than predicted on the basis of physiology, but was unable to fully document the expressed opsin genes responsible for this diversity. Furthermore, questions remain about how other components of phototransduction cascades are involved. This study continues prior investigations by examining the molecular function of stomatopods' visual systems using new whole eye 454 transcriptome datasets from two species, Hemisquilla californiensis and Pseudosquilla ciliata. These two species represent taxonomic diversity within the order Stomatopoda, as well as variations in the anatomy and physiology of the visual system. Using an evolutionary placement algorithm to annotate the transcriptome, we identified the presence of nine components of the stomatopods' G-protein-coupled receptor (GPCR) phototransduction cascade, including two visual arrestins, subunits of the heterotrimeric G-protein, phospholipase C, transient receptor potential channels, and opsin transcripts. The set of expressed transduction genes suggests that stomatopods utilize a Gq-mediated GPCR-signaling cascade. The most notable difference in expression between the phototransduction cascades of the two species was the number of opsin contigs recovered, with 18 contigs found in retinas of H. californiensis, and 49 contigs in those of P. ciliata. Based on phylogenetic placement and fragment overlap, these contigs were estimated to represent 14 and 33 expressed transcripts, respectively. These data expand the known opsin diversity in stomatopods to clades of arthropod opsins that are sensitive to short wavelengths and ultraviolet wavelengths and confirm the results of previous studies recovering more opsin transcripts than spectrally distinct types of photoreceptors. Many of the recovered transcripts were phylogenetically placed in an evolutionary clade of crustacean opsin sequences that is rapidly expanding as the visual systems from more species are investigated. We discuss these results in relation to the emerging pattern, particularly in crustacean visual systems, of the expression of multiple opsin transcripts in photoreceptors of the same spectral class, and even in single photoreceptor cells. PMID:23727979

  12. Eye-independent, light-activated chromatophore expansion (LACE) and expression of phototransduction genes in the skin of Octopus bimaculoides

    PubMed Central

    Ramirez, M. Desmond; Oakley, Todd H.

    2015-01-01

    ABSTRACT Cephalopods are renowned for changing the color and pattern of their skin for both camouflage and communication. Yet, we do not fully understand how cephalopods control the pigmented chromatophore organs in their skin and change their body pattern. Although these changes primarily rely on eyesight, we found that light causes chromatophores to expand in excised pieces of Octopus bimaculoides skin. We call this behavior light-activated chromatophore expansion (or LACE). To uncover how octopus skin senses light, we used antibodies against r-opsin phototransduction proteins to identify sensory neurons that express r-opsin in the skin. We hypothesized that octopus LACE relies on the same r-opsin phototransduction cascade found in octopus eyes. By creating an action spectrum for the latency to LACE, we found that LACE occurred most quickly in response to blue light. We fit our action spectrum data to a standard opsin curve template and estimated the λmax of LACE to be 480 nm. Consistent with our hypothesis, the maximum sensitivity of the light sensors underlying LACE closely matches the known spectral sensitivity of opsin from octopus eyes. LACE in isolated preparations suggests that octopus skin is intrinsically light sensitive and that this dispersed light sense might contribute to their unique and novel patterning abilities. Finally, our data suggest that a common molecular mechanism for light detection in eyes may have been co-opted for light sensing in octopus skin and then used for LACE. PMID:25994633

  13. Optogenetics for retinal disorders.

    PubMed

    Henriksen, Bradley S; Marc, Robert E; Bernstein, Paul S

    2014-01-01

    Optogenetics is the use of genetic methods combined with optical technology to achieve gain or loss of function within neuronal circuits. The field of optogenetics has been rapidly expanding in efforts to restore visual function to blinding diseases such as retinitis pigmentosa (RP). Most work in the field includes a group of light-sensitive retinaldehyde-binding proteins known as opsins. Opsins couple photon absorption to molecular signaling chains that control cellular ion currents. Targeting of opsin genes to surviving retinal cells is fundamental to the success of optogenetic therapy. Viral delivery, primarily adeno-associated virus, using intravitreal injection for inner retinal cells and subretinal injection for outer retinal cells, has proven successful in many models. Challenges in bioengineering remain for optogenetics including relative insensitivity of opsins to physiologic light levels of stimulation and difficulty with viral delivery in primate models. However, targeting optogenetic therapy may present an even greater challenge. Neural and glial remodeling seen in advanced stages of RP result in reorganization of remaining neural retina, and optogenetic therapy may not yield functional results. Remodeling also poses a challenge to the selection of cellular targets, with bipolar, amacrine and ganglion cells all playing distinct physiologic roles, and affected by remodeling differently. Although optogenetics has drawn closer to clinical utility, advances in opsin engineering, therapeutic targeting and ultimately in molecular inhibition of remodeling will play critical roles in the continued clinical advancement of optogenetic therapy. PMID:25667740

  14. The eyes have it: regulatory and structural changes both underlie cichlid visual pigment diversity.

    PubMed

    Hofmann, Christopher M; O'Quin, Kelly E; Marshall, N Justin; Cronin, Thomas W; Seehausen, Ole; Carleton, Karen L

    2009-12-01

    A major goal of evolutionary biology is to unravel the molecular genetic mechanisms that underlie functional diversification and adaptation. We investigated how changes in gene regulation and coding sequence contribute to sensory diversification in two replicate radiations of cichlid fishes. In the clear waters of Lake Malawi, differential opsin expression generates diverse visual systems, with sensitivities extending from the ultraviolet to the red regions of the spectrum. These sensitivities fall into three distinct clusters and are correlated with foraging habits. In the turbid waters of Lake Victoria, visual sensitivity is constrained to longer wavelengths, and opsin expression is correlated with ambient light. In addition to regulatory changes, we found that the opsins coding for the shortest- and longest-wavelength visual pigments have elevated numbers of potentially functional substitutions. Thus, we present a model of sensory evolution in which both molecular genetic mechanisms work in concert. Changes in gene expression generate large shifts in visual pigment sensitivity across the collective opsin spectral range, but changes in coding sequence appear to fine-tune visual pigment sensitivity at the short- and long-wavelength ends of this range, where differential opsin expression can no longer extend visual pigment sensitivity. PMID:20027211

  15. Eye-independent, light-activated chromatophore expansion (LACE) and expression of phototransduction genes in the skin of Octopus bimaculoides.

    PubMed

    Ramirez, M Desmond; Oakley, Todd H

    2015-05-15

    Cephalopods are renowned for changing the color and pattern of their skin for both camouflage and communication. Yet, we do not fully understand how cephalopods control the pigmented chromatophore organs in their skin and change their body pattern. Although these changes primarily rely on eyesight, we found that light causes chromatophores to expand in excised pieces of Octopus bimaculoides skin. We call this behavior light-activated chromatophore expansion (or LACE). To uncover how octopus skin senses light, we used antibodies against r-opsin phototransduction proteins to identify sensory neurons that express r-opsin in the skin. We hypothesized that octopus LACE relies on the same r-opsin phototransduction cascade found in octopus eyes. By creating an action spectrum for the latency to LACE, we found that LACE occurred most quickly in response to blue light. We fit our action spectrum data to a standard opsin curve template and estimated the λmax of LACE to be 480 nm. Consistent with our hypothesis, the maximum sensitivity of the light sensors underlying LACE closely matches the known spectral sensitivity of opsin from octopus eyes. LACE in isolated preparations suggests that octopus skin is intrinsically light sensitive and that this dispersed light sense might contribute to their unique and novel patterning abilities. Finally, our data suggest that a common molecular mechanism for light detection in eyes may have been co-opted for light sensing in octopus skin and then used for LACE. PMID:25994633

  16. Absorbance spectra and molecular structure of the blue-sensitive rod visual pigment in the conger eel (Conger conger).

    PubMed

    Archer, S; Hirano, J

    1996-06-22

    The conger eel (Conger conger) is a nocturnal fish that can be found living in shallow coastal water and deep water down to 1000 m. The conger eel has a pure rod retina with a visual pigment maximally sensitive to blue light around 487 nm. We have cloned and sequenced the opsin cDNA which is presumed to code for this visual pigment and have found it to be highly homologous to the form of opsin that is expressed in mature, deep-living European eels (Anguilla anguilla). The opsin sequence information presented here provides additional evidence that specific amino acid sites are involved in the spectral tuning of this class of blue-sensitive visual pigments. PMID:8763796

  17. Binding specificity of retinal analogs to photoactivated visual pigments suggest mechanism for fine-tuning GPCR-ligand interactions.

    PubMed

    Srinivasan, Sundaramoorthy; Ramon, Eva; Cordomí, Arnau; Garriga, Pere

    2014-03-20

    11-cis-retinal acts as an inverse agonist stabilizing the inactive conformation of visual pigments, and upon photoactivation, it isomerizes to all-trans-retinal, initiating signal transduction. We have analyzed opsin regeneration with retinal analogs for rhodopsin and red cone opsin. We find differential binding of the analogs to the receptors after photobleaching and a dependence of the binding kinetics on the oligomerization state of the protein. The results outline the sensitivity of retinal entry to the binding pocket of visual receptors to the specific conformation adopted by the receptor and by the molecular architecture defined by specific amino acids in the binding pocket and the retinal entry site, as well as the topology of the retinal analog. Overall, our findings highlight the specificity of the ligand-opsin interactions, a feature that can be shared by other G-protein-coupled receptors. PMID:24560606

  18. Cone visual pigments.

    PubMed

    Imamoto, Yasushi; Shichida, Yoshinori

    2014-05-01

    Cone visual pigments are visual opsins that are present in vertebrate cone photoreceptor cells and act as photoreceptor molecules responsible for photopic vision. Like the rod visual pigment rhodopsin, which is responsible for scotopic vision, cone visual pigments contain the chromophore 11-cis-retinal, which undergoes cis-trans isomerization resulting in the induction of conformational changes of the protein moiety to form a G protein-activating state. There are multiple types of cone visual pigments with different absorption maxima, which are the molecular basis of color discrimination in animals. Cone visual pigments form a phylogenetic sister group with non-visual opsin groups such as pinopsin, VA opsin, parapinopsin and parietopsin groups. Cone visual pigments diverged into four groups with different absorption maxima, and the rhodopsin group diverged from one of the four groups of cone visual pigments. The photochemical behavior of cone visual pigments is similar to that of pinopsin but considerably different from those of other non-visual opsins. G protein activation efficiency of cone visual pigments is also comparable to that of pinopsin but higher than that of the other non-visual opsins. Recent measurements with sufficient time-resolution demonstrated that G protein activation efficiency of cone visual pigments is lower than that of rhodopsin, which is one of the molecular bases for the lower amplification of cones compared to rods. In this review, the uniqueness of cone visual pigments is shown by comparison of their molecular properties with those of non-visual opsins and rhodopsin. This article is part of a Special Issue entitled: Retinal Proteins - You can teach an old dog new tricks. PMID:24021171

  19. Spectral and temporal sensitivity of cone-mediated responses in mouse retinal ganglion cells

    PubMed Central

    Wang, Yanbin V.; Weick, Michael; Demb, Jonathan B.

    2011-01-01

    The retina uses two photoreceptor types to encode the wide range of light intensities in the natural environment. Rods mediate vision in dim light, whereas cones mediate vision in bright light. Mouse photoreceptors include only 3% cones, and the majority of these co-express two opsins (S, M), with peak sensitivity to either ultraviolet (360 nm) or green light (508 nm). The M:S opsin ratio varies across the retina but has not been characterized functionally, preventing quantitative study of cone-mediated vision. Furthermore, physiological and behavioral measurements suggested that mouse retina supports relatively slow temporal processing (peak sensitivity, ~2–5 Hz), compared to primates; however, past studies used visible wavelengths that are inefficient at stimulating mouse S opsin. Here, we measured the M:S opsin expression ratio across the mouse retina, as reflected by ganglion cell responses, in vitro, and probed cone-mediated ganglion cell temporal properties using ultraviolet light stimulation and linear systems analysis. From recordings in mice lacking rod function (Gnat1−/−, Rho−/−), we estimate ~70% M-opsin expression in far dorsal retina, dropping to <5% M-opsin expression throughout ventral retina. In mice lacking cone function (Gnat2cpfl3), light-adapted rod-mediated responses peaked at ~5–7 Hz. In wild-type mice, cone-mediated responses peaked at ~10 Hz, with substantial responsiveness up to ~30 Hz. Therefore, despite the small percentage of cones, cone-mediated responses in mouse ganglion cells are fast and robust, similar to those in primates. These measurements enable quantitative analysis of cone-mediated responses at all levels of the visual system. PMID:21613480

  20. Molecular evidence for color discrimination in the Atlantic sand fiddler crab, Uca pugilator.

    PubMed

    Rajkumar, Premraj; Rollmann, Stephanie M; Cook, Tiffany A; Layne, John E

    2010-12-15

    Fiddler crabs are intertidal brachyuran crabs that belong to the genus Uca. Approximately 97 different species have been identified, and several of these live sympatrically. Many have species-specific body color patterns that may act as signals for intra- and interspecific communication. To understand the behavioral and ecological role of this coloration we must know whether fiddler crabs have the physiological capacity to perceive color cues. Using a molecular approach, we identified the opsin-encoding genes and determined their expression patterns across the eye of the sand fiddler crab, Uca pugilator. We identified three different opsin-encoding genes (UpRh1, UpRh2 and UpRh3). UpRh1 and UpRh2 are highly related and have similarities in their amino acid sequences to other arthropod long- and medium-wavelength-sensitive opsins, whereas UpRh3 is similar to other arthropod UV-sensitive opsins. All three opsins are expressed in each ommatidium, in an opsin-specific pattern. UpRh3 is present only in the R8 photoreceptor cell, whereas UpRh1 and UpRh2 are present in the R1-7 cells, with UpRh1 expression restricted to five cells and UpRh2 expression present in three cells. Thus, one photoreceptor in every ommatidium expresses both UpRh1 and UpRh2, providing another example of sensory receptor coexpression. These results show that U. pugilator has the basic molecular machinery for color perception, perhaps even trichromatic vision. PMID:21113005

  1. Molecular evidence for color discrimination in the Atlantic sand fiddler crab, Uca pugilator

    PubMed Central

    Rajkumar, Premraj; Rollmann, Stephanie M.; Cook, Tiffany A.; Layne, John E.

    2010-01-01

    SUMMARY Fiddler crabs are intertidal brachyuran crabs that belong to the genus Uca. Approximately 97 different species have been identified, and several of these live sympatrically. Many have species-specific body color patterns that may act as signals for intra- and interspecific communication. To understand the behavioral and ecological role of this coloration we must know whether fiddler crabs have the physiological capacity to perceive color cues. Using a molecular approach, we identified the opsin-encoding genes and determined their expression patterns across the eye of the sand fiddler crab, Uca pugilator. We identified three different opsin-encoding genes (UpRh1, UpRh2 and UpRh3). UpRh1 and UpRh2 are highly related and have similarities in their amino acid sequences to other arthropod long- and medium-wavelength-sensitive opsins, whereas UpRh3 is similar to other arthropod UV-sensitive opsins. All three opsins are expressed in each ommatidium, in an opsin-specific pattern. UpRh3 is present only in the R8 photoreceptor cell, whereas UpRh1 and UpRh2 are present in the R1-7 cells, with UpRh1 expression restricted to five cells and UpRh2 expression present in three cells. Thus, one photoreceptor in every ommatidium expresses both UpRh1 and UpRh2, providing another example of sensory receptor coexpression. These results show that U. pugilator has the basic molecular machinery for color perception, perhaps even trichromatic vision. PMID:21113005

  2. Expression of developmentally defined retinal phenotypes in the histogenesis of retinoblastoma.

    PubMed Central

    Gonzalez-Fernandez, F.; Lopes, M. B.; Garcia-Fernandez, J. M.; Foster, R. G.; De Grip, W. J.; Rosemberg, S.; Newman, S. A.; VandenBerg, S. R.

    1992-01-01

    Retinoblastoma, the most common intraocular tumor of childhood, is a malignant neoplasm that arises during retinal development. The embryonal cell target for neoplastic transformation is not yet clearly defined. To better understand the histogenetic potential of this tumor, the expression of photoreceptor and glial cell-associated proteins were examined in 22 primary retinoblastomas. Interphotoreceptor retinol-binding protein (IRBP), cone and rod opsins were selected as the photoreceptor specific proteins due to their different temporal patterns of expression during normal retinal development. Neoplastic Müller cell differentiation, and non-neoplastic reactive astrocytes were identified using cellular retinaldehyde binding-protein (CRAlBP), and glial fibrillary acidic protein (GFAP), respectively. Photoreceptor proteins were present in 16 cases and showed different cellular patterns of expression. IRBP and cone opsin were usually abundant. Although rod opsin was clearly identified in eight tumors, its expression was more restricted than either IRBP or cone opsin. This differential pattern of expression, opposite to the normal pattern of photoreceptor gene expression in the adult retina, corresponded to a marked decrease in mRNA for rod opsin. Cone opsin and IRBP colocalized in fleurettes demonstrating that neoplastic human cone cells are capable of IRBP synthesis. Müller cell differentiation was present in 12 of the 16 cases in which photoreceptor proteins were detected. In contrast, GFAP was only present in reactive, stromal astrocytes associated with blood vessels. Our data suggest that the retinoblastoma has the histogenetic potential of the immature neural retinal epithelium which can give rise to both photoreceptor and Müller cell lineages. The differential expression of cone and rod phenotypes in retinoblastoma is consistent with the "default" mechanism of cone cell differentiation. Images Figure 7 Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:1386715

  3. PyRhO: A Multiscale Optogenetics Simulation Platform

    PubMed Central

    Evans, Benjamin D.; Jarvis, Sarah; Schultz, Simon R.; Nikolic, Konstantin

    2016-01-01

    Optogenetics has become a key tool for understanding the function of neural circuits and controlling their behavior. An array of directly light driven opsins have been genetically isolated from several families of organisms, with a wide range of temporal and spectral properties. In order to characterize, understand and apply these opsins, we present an integrated suite of open-source, multi-scale computational tools called PyRhO. The purpose of developing PyRhO is three-fold: (i) to characterize new (and existing) opsins by automatically fitting a minimal set of experimental data to three-, four-, or six-state kinetic models, (ii) to simulate these models at the channel, neuron and network levels, and (iii) provide functional insights through model selection and virtual experiments in silico. The module is written in Python with an additional IPython/Jupyter notebook based GUI, allowing models to be fit, simulations to be run and results to be shared through simply interacting with a webpage. The seamless integration of model fitting algorithms with simulation environments (including NEURON and Brian2) for these virtual opsins will enable neuroscientists to gain a comprehensive understanding of their behavior and rapidly identify the most suitable variant for application in a particular biological system. This process may thereby guide not only experimental design and opsin choice but also alterations of the opsin genetic code in a neuro-engineering feed-back loop. In this way, we expect PyRhO will help to significantly advance optogenetics as a tool for transforming biological sciences. PMID:27148037

  4. Phototransduction Motifs and Variations

    PubMed Central

    Yau, King-Wai; Hardie, Roger C.

    2010-01-01

    Seeing begins in the photoreceptors, where light is absorbed and signaled to the nervous system. Throughout the animal kingdom, photoreceptors are diverse in design and purpose. Nonetheless, phototransduction—the mechanism by which absorbed photons are converted into an electrical response—is highly conserved and based almost exclusively on a single class of photoproteins, the opsins. In this Review, we survey the G protein-coupled signaling cascades downstream from opsins in photoreceptors across vertebrate and invertebrate species, noting their similarities as well as differences. PMID:19837030

  5. Comprehensive analysis of escape-cone losses from luminescent waveguides.

    PubMed

    McDowall, Stephen; Butler, Tristan; Bain, Edward; Scharnhorst, Kelsey; Patrick, David

    2013-02-20

    Luminescent waveguides (LWs) occur in a wide range of applications, from solar concentrators to doped fiber amplifiers. Here we report a comprehensive analysis of escape-cone losses in LWs, which are losses associated with internal rays making an angle less than the critical angle with a waveguide surface. For applications such as luminescent solar concentrators, escape-cone losses often dominate all others. A statistical treatment of escape-cone losses is given accounting for photoselection, photon polarization, and the Fresnel relations, and the model is used to analyze light absorption and propagation in waveguides with isotropic and orientationally aligned luminophores. The results are then compared to experimental measurements performed on a fluorescent dye-doped poly(methyl methacrylate) waveguide. PMID:23434994

  6. Effect of 24 months of recombinant growth hormone on height and body proportions in SHOX haploinsufficiency.

    PubMed

    Munns, C F J; Berry, M; Vickers, D; Rappold, G A; Hyland, V J; Glass, I A; Batch, J A

    2003-09-01

    Leri-Weill syndrome (LWS) is a skeletal dysplasia with mesomelic short stature, bilateral Madelung deformity (BMD) and SHOX (short stature homeobox-containing gene) haploinsufficiency. The effect of 24 months of recombinant human growth hormone (rhGH) therapy on the stature and BMD of two females with SHOX haploinsufficiency (demonstrated by fluorescence in situ hybridisation) and LWS was evaluated. Both patients demonstrated an increase in height standard deviation score (SDS) and height velocity SDS over the 24 months of therapy. Patient 1 demonstrated a relative increase in arm-span and upper segment measurements with rhGH while patient 2 demonstrated a relative increase in lower limb length. There was appropriate advancement of bone age, no adverse events and no significant deterioration in BMD. In this study, 24 months of rhGH was a safe and effective therapy for the disproportionate short stature of SHOX haploinsufficiency, with no clinical deterioration of BMD. PMID:14513876

  7. NASA's Living with a Star Program: The Geospace Mission Concept

    NASA Technical Reports Server (NTRS)

    Barth, Janet L.; Giles, Barbara; Zanetti, Lawrence; Spann, James; Day, John H. (Technical Monitor)

    2002-01-01

    NASA has initiated the Living with a Star Program (LWS) to develop the scientific understanding to address the aspects of the Connected Sun-Earth system that affect life and society. A goal of the program is to bridge the gap between science, engineering, and user application communities. This will enable future science, operational, and commercial objectives in space and atmospheric environments by improving engineering approaches to the accommodation and/or mitigation of the effects of solar variability on technological systems. Three program elements are the Science Missions; a Theory, Modeling, and Data Analysis program; and a Space Environment Testbeds program. Because many of the effects of solar variability on humanity are observed in Geospace regions of space, the science research for all three elements of the LWS Program have significant components in Geospace regions.

  8. The Stroop effect in English-Japanese bilinguals: the effect of phonological similarity.

    PubMed

    Sumiya, Hiromi; Healy, Alice F

    2008-01-01

    English-Japanese bilinguals performed a Stroop color-word interference task with both English and Japanese stimuli and responded in both English and Japanese. The Japanese stimuli were either the traditional color terms (TCTs) written in Hiragana or loanwords (LWs) from English written in Katakana. Both within-language and between-language interference were found for all combinations of stimuli and responses. The between-language interference was larger for Katakana LWs (phonologically similar to English) than for Hiragana TCTs, especially with Japanese responses. The magnitude of this phonological effect increased with self-rated reading fluency in Japanese. Overall responding was slower and the Stroop effect larger with English than with Japanese stimuli. These results suggest that unintentional lexical access elicits automatic phonological processing even with intermediate-level reading proficiency. PMID:18444519

  9. Solar Dynamics Observatory and Solar Sentinels Missions in Living With A Star Program

    NASA Astrophysics Data System (ADS)

    Guhathakurta, M.

    We live in the extended atmosphere of an active star. While sunlight enables and sustains life, the Sun's variability produces streams of high-energy particles and radiation that can affect life. In the year 2000 NASA established a new program called the Living With a Star (LWS). The objective of this program is to "develop the scientific understanding necessary to effectively address those aspects of the Connected SunEarth system that directly affect life and society." To address this objective NASA formulated two groups of mission scenarios in the LWS Space Weather Research Network: (a) solar dynamics elements (Solar Dynamics Observatory/Sentinels) that observe the Sun and track disturbances originating there and (b) geospace dynamics elements (Geospace Missions Network) consisting of constellations of small satellites located in key regions around the Earth to measure downstream effects. Science payload for the first mission of LWS, namely Solar Dynamics Observatory (SDO) to study the Sun as a magnetic variable star and quantify its input into the Earth's atmosphere has been selected with a launch date of 2007. Details of the scientific objectives of the mission and its payload will be discussed. Solar Sentinels, a set of notional missions to investigate the physics of interplanetary medium that link solar processes to Earth's environment are planned to be launched in the 2010-2013 time frame depending on budgetary and other factors. A small fleet of spacecraft (four in the nominal plan) will be used to track and understand the evolution of matter ejected from the Sun. Concepts which are presently under study for the future missions will be described in detail. Although originally a U.S. initiative, the problems that LWS addresses are global in nature. Thus under the auspices of IACG an International Living With a Star (ILWS) working group has been established and I will report on the joint activities between NASA and other space agencies.

  10. Evaluation of mercury in the liquid waste processing facilities

    SciTech Connect

    Jain, Vijay; Shah, Hasmukh; Occhipinti, John E.; Wilmarth, William R.; Edwards, Richard E.

    2015-08-13

    This report provides a summary of Phase I activities conducted to support an Integrated Evaluation of Mercury in Liquid Waste System (LWS) Processing Facilities. Phase I activities included a review and assessment of the liquid waste inventory and chemical processing behavior of mercury using a system by system review methodology approach. Gaps in understanding mercury behavior as well as action items from the structured reviews are being tracked. 64% of the gaps and actions have been resolved.

  11. Evaluation of mercury in liquid waste processing facilities - Phase I report

    SciTech Connect

    Jain, V.; Occhipinti, J. E.; Shah, H.; Wilmarth, W. R.; Edwards, R. E.

    2015-07-01

    This report provides a summary of Phase I activities conducted to support an Integrated Evaluation of Mercury in Liquid Waste System (LWS) Processing Facilities. Phase I activities included a review and assessment of the liquid waste inventory and chemical processing behavior of mercury using a system by system review methodology approach. Gaps in understanding mercury behavior as well as action items from the structured reviews are being tracked. 64% of the gaps and actions have been resolved.

  12. Evaluation of Mercury in Liquid Waste Processing Facilities - Phase I Report

    SciTech Connect

    Jain, V.; Occhipinti, J.; Shah, H.; Wilmarth, B.; Edwards, R.

    2015-07-01

    This report provides a summary of Phase I activities conducted to support an Integrated Evaluation of Mercury in Liquid Waste System (LWS) Processing Facilities. Phase I activities included a review and assessment of the liquid waste inventory and chemical processing behavior of mercury using a system by system review methodology approach. Gaps in understanding mercury behavior as well as action items from the structured reviews are being tracked. 64% of the gaps and actions have been resolved.

  13. Wetland treatment of oil and gas well wastewaters. Quarterly technical report, November 25, 1993--March 24, 1994

    SciTech Connect

    Kadlec, R.H.; Srinivasan, K.R.

    1994-04-15

    In the third quarterly report, adsorption of heavy metals ions such as Cu(II) and Cr(VI) onto soils drawn from the laboratory-type wetland (LW) was shown to be weak. On the other hand, it was shown that modified-clays did adsorb Cr(VI) ions strongly at pH 4.5. Further, studies on the pH dependence of the adsorption of {beta}-naphthoic acid, (NA), a well-documented contaminant in many oil and gas well waste waters (4), onto modified-clays were undertaken and it was shown that uptake of NA by modified-clays was of the high affinity type at pH 4.5 and 7.0, but weak at pH 9.0. Adsorption of heavy metal ions, Cu{sup 2+}, and CR(VI) onto algae, a proposed wetland amendment, was carried out and the results were presented and discussed in the fourth quarterly report. Studies on the dynamics of uptake of phenol and NA by laboratory-type wetlands (LWs) were initiated and preliminary results indicated that both phenol and NA were sorbed onto components of LWs. A mass balance model has been developed to quantify the fate of phenol in LWs. The model is based on the postulate that the fate of phenol in LWs can be attributed to a combination of (1) evaporation of solute and solvent, (2) adsorption of phenol onto various components of LW and (3) its biodegradation, both in solution and at solid-liquid interface. As an initial approximation, the latter two processes have been lumped together and incorporated into the model as an unit operation. Both zero order and first order kinetics for the disappearance of phenol have been considered. Evaporative losses of water and phenol have also been taken into account and this model is presented and discussed in this quarterly report.

  14. Delayed access of low body weight-selected chicks to food at hatch is associated with up-regulated pancreatic glucagon and glucose transporter gene expression.

    PubMed

    Parker, Grace A; Sumners, Lindsay H; Zhao, Xiaoling; Honaker, Christa F; Siegel, Paul B; Cline, Mark A; Gilbert, Elizabeth R

    2015-11-01

    Chickens selected for low (LWS) and high (HWS) juvenile body weight (BW) for 55 generations differ in BW by 10-fold at selection age. High (HWR) and low (LWR) body weight-relaxed lines have been random-bred since the 46th generation. Our objective was to evaluate the developmental and nutritional regulation of pancreatic mRNA abundance of pancreatic and duodenal homeobox 1 (PDX1), preproinsulin (PPI), preproglucagon (PPG), and glucose transporter 2 (GLUT2). At day of hatch (DOH) and days 1, 3, 7, and 15 (D1, 3, 7 and 15, respectively), pancreas was collected and real time PCR was performed in Experiment 1. In Experiment 2, HWS and LWS were fed or delayed access to food for 72 h post-hatch, and pancreas collected at D15. There was an interaction of line and age for GLUT2 (P=0.001), PPI (P<0.0001), PPG (P=0.034), and PDX1 (P<0.0001). Expression was greater in chicks from LWR and LWS than HWR and HWS. There was an interaction of line and nutrition on PPG (P<0.0001) and GLUT2 (P=0.001) mRNA, where expression was similar among chicks that were fed but greater in LWS than HWS when chicks were delayed access to food. Thus, the first two weeks is important for maturation of pancreatic endocrine function. Long-term selection for BW is associated with differences in pancreas development, and delaying access to food at hatch may have persisting effects on glucose regulatory function. PMID:26260898

  15. The Living With a Star CDAW on the Solar and Geospace Connections of Solar Energetic Particles

    NASA Technical Reports Server (NTRS)

    Thompson, Barbara J.; Gopalswamy, Nat; Colon, Gilberto (Technical Monitor)

    2002-01-01

    The Living With a Star Program is sponsoring its first CDAW (Coordinated Data Analysis Workshop) to be held July 23-26, 2002 at a conference support location near the NASA Goddard Space Flight Center. This CDAW's topic is Solar Energetic Particle events. The topic was chosen due to the breadth of the impact of SEP's on the space environment and terrestrial climate. General goals of the LWS CDAW are a) Stimulate LWS Science on the near term, b) Facilitate cross-disciplinary interaction between the LWS scientific and space environment communities, c) Produce science products for all potential users, and d) Assist in the development of the LWS data system. The workshop will proceed similar to a previous CDAW held in 1999 on Interplanetary Type 11 Shocks. A list of target events has been compiled, which can be found at the workshop home page. The page lists all of the SEP events from 1996 January to 2001 December with energy > 10 MeV particle intensities exceeding 10 PFU. Preparation for the workshop consists of identifying relevant data from a wide variety of sources (solar, interplanetary, magnetospheric and climatary), accumulating the data (frequently this consists of both raw data, processed data and plots to ease perusal during the workshop) and gathering the software tools. Participants in the workshop are expected to complete their contributions of data or models prior to arriving at the workshop. Most of the CDAW consists of joint analysis of this data; only a few introductory talks are given at the beginning of the workshop, with the rest of the time being devoted to producing scientific results. Additional symposia may be scheduled at a later date, which will allow a venue for scientific talks on the CDAW results and associated science. The poster will list the scientific goals of the workshop, as well as a scientific discussion of the data which has been accumulated thus far.

  16. Implementing a Virtual Workshop for Interdiscplinary collaboration on Grand Challenge Issues: Lessons Learned

    NASA Astrophysics Data System (ADS)

    Barnes, R. J.; Kozyra, J.; Weiss, M.; Fox, N. J.; Morrison, D.; Paxton, L.

    2006-12-01

    During October of 2006 a Virtual Workshop (eWorkshop), sponsored by CAWSES, NASA/LWS, eGY, IHY, NSF, and ICESTAR was held to discuss the state of the Sun-Earth system during super substorms. This virtual workshop used the Internet to allow world-wide participants to discuss and exchange data using various web based tools. Presented here are the technical aspects of the implementation and the lessons learned from this preliminary workshop.

  17. Proviral integrations and expression of endogenous Avian leucosis virus during long term selection for high and low body weight in two chicken lines

    PubMed Central

    Ka, Sojeong; Kerje, Susanne; Bornold, Lina; Liljegren, Ulrika; Siegel, Paul B; Andersson, Leif; Hallböök, Finn

    2009-01-01

    Background Long-term selection (> 45 generations) for low or high juvenile body weight from a common founder population of White Plymouth Rock chickens has generated two extremely divergent lines, the LWS and HWS lines. In addition to a > 9-fold difference between lines for the selected trait, large behavioural and metabolic differences between the two lines evolved during the course of the selection. We recently compared gene expression in brain tissue from birds representing these lines using a global cDNA array analysis and the results showed multiple but small expression differences in protein coding genes. The main differentially expressed transcripts were endogenous retroviral sequences identified as avian leucosis virus subgroup-E (ALVE). Results In this work we confirm the differential ALVE expression and analysed expression and number of proviral integrations in the two parental lines as well as in F9 individuals from an advanced intercross of the lines. Correlation analysis between expression, proviral integrations and body weight showed that high ALVE levels in the LWS line were inherited and that more ALVE integrations were detected in LWS than HWS birds. Conclusion We conclude that only a few of the integrations contribute to the high expression levels seen in the LWS line and that high ALVE expression was significantly correlated with lower body weights for the females but not males. The conserved correlation between high expression and low body weight in females after 9 generations of intercrosses, indicated that ALVE loci conferring high expression directly affects growth or are very closely linked to loci regulating growth. PMID:19604406

  18. Assessing Lvy walks as models of animal foraging

    PubMed Central

    James, Alex; Plank, Michael J.; Edwards, Andrew M.

    2011-01-01

    The hypothesis that the optimal search strategy is a Lvy walk (LW) or Lvy flight, originally suggested in 1995, has generated an explosion of interest and controversy. Long-standing empirical evidence supporting the LW hypothesis has been overturned, while new models and data are constantly being published. Statistical methods have been criticized and new methods put forward. In parallel with the empirical studies, theoretical search models have been developed. Some theories have been disproved while others remain. Here, we gather together the current state of the art on the role of LWs in optimal foraging theory. We examine the body of theory underpinning the subject. Then we present new results showing that deviations from the idealized one-dimensional search model greatly reduce or remove the advantage of LWs. The search strategy of an LW with exponent ? = 2 is therefore not as robust as is widely thought. We also review the available techniques, and their potential pitfalls, for analysing field data. It is becoming increasingly recognized that there is a wide range of mechanisms that can lead to the apparent observation of power-law patterns. The consequence of this is that the detection of such patterns in field data implies neither that the foragers in question are performing an LW, nor that they have evolved to do so. We conclude that LWs are neither a universal optimal search strategy, nor are they as widespread in nature as was once thought. PMID:21632609

  19. Leptin effects on food and water intake in lines of chickens selected for high or low body weight.

    PubMed

    Kuo, Alice Y; Cline, Mark A; Werner, Elizabeth; Siegel, Paul B; Denbow, D Michael

    2005-03-16

    There is an association between autonomic nervous system output and obesity. The sympathetic nervous system stimulates lipid metabolism and regulates food intake and, hence, body weight. Leptin, produced by adipocytes in proportion to their size, has been shown to directly stimulate the satiety center. In the experiment reported here, food and water intake were compared after intracerebroventricular administration of human recombinant leptin to lines of chickens that had undergone divergent selection for over 45 generations from a common White Rock base population for high (HWS) or low (LWS) body weight at 8 weeks-of-age. Leptin caused a linear decrease in food intake in chickens from the LWS line whereas no effect was observed in those from the HWS line. The HWS chickens tended to have reduced water intake post leptin administration. Others reported that leptin decreased food intake in both broiler and Leghorn chickens. Leptin concentration in the central nervous system may not contribute directly to the difference of body weight between HWS and LWS chickens. PMID:15763584

  20. Solar Sentinels: Report of the Science and Technology Definition Team

    NASA Technical Reports Server (NTRS)

    2006-01-01

    The goal of NASA s Living With a Star (LWS) program is to develop the scientific understanding necessary to effectively address those aspects of the connected Sun Earth system that directly affect life and society. Along with the other elements of LWS, Solar Sentinels aims to discover, understand, and model the heliospheric initiation, propagation, and solar connection of those energetic phenomena that adversely affect space exploration and life and society here on Earth. The Solar Sentinels mission will address the following questions: (1) How, where, and under what circumstances are solar energetic particles (SEPs) accelerated to high energies and how do they propagate through the heliosphere? And (2) How are solar wind structures associated with these SEPs, like CMEs, shocks, and high-speed streams, initiated, propagate, evolve, and interact in the inner heliosphere? The Sentinels STDT recommends implementing this mission in two portions, one optimized for inner heliospheric in-situ measurements and the other for solar remote observations. Sentinels will greatly enhance the overall LWS science return.

  1. Multi-Mode Electromagnetic Ultrasonic Lamb Wave Tomography Imaging for Variable-Depth Defects in Metal Plates.

    PubMed

    Huang, Songling; Zhang, Yu; Wang, Shen; Zhao, Wei

    2016-01-01

    This paper proposes a new cross-hole tomography imaging (CTI) method for variable-depth defects in metal plates based on multi-mode electromagnetic ultrasonic Lamb waves (LWs). The dispersion characteristics determine that different modes of LWs are sensitive to different thicknesses of metal plates. In this work, the sensitivities to thickness variation of A0- and S0-mode LWs are theoretically studied. The principles and procedures for the cooperation of A0- and S0-mode LW CTI are proposed. Moreover, the experimental LW imaging system on an aluminum plate with a variable-depth defect is set up, based on A0- and S0-mode EMAT (electromagnetic acoustic transducer) arrays. For comparison, the traditional single-mode LW CTI method is used in the same experimental platform. The imaging results show that the computed thickness distribution by the proposed multi-mode method more accurately reflects the actual thickness variation of the defect, while neither the S0 nor the A0 single-mode method was able to distinguish thickness variation in the defect region. Moreover, the quantification of the defect's thickness variation is more accurate with the multi-mode method. Therefore, theoretical and practical results prove that the variable-depth defect in metal plates can be successfully quantified and visualized by the proposed multi-mode electromagnetic ultrasonic LW CTI method. PMID:27144571

  2. Multi-Mode Electromagnetic Ultrasonic Lamb Wave Tomography Imaging for Variable-Depth Defects in Metal Plates

    PubMed Central

    Huang, Songling; Zhang, Yu; Wang, Shen; Zhao, Wei

    2016-01-01

    This paper proposes a new cross-hole tomography imaging (CTI) method for variable-depth defects in metal plates based on multi-mode electromagnetic ultrasonic Lamb waves (LWs). The dispersion characteristics determine that different modes of LWs are sensitive to different thicknesses of metal plates. In this work, the sensitivities to thickness variation of A0- and S0-mode LWs are theoretically studied. The principles and procedures for the cooperation of A0- and S0-mode LW CTI are proposed. Moreover, the experimental LW imaging system on an aluminum plate with a variable-depth defect is set up, based on A0- and S0-mode EMAT (electromagnetic acoustic transducer) arrays. For comparison, the traditional single-mode LW CTI method is used in the same experimental platform. The imaging results show that the computed thickness distribution by the proposed multi-mode method more accurately reflects the actual thickness variation of the defect, while neither the S0 nor the A0 single-mode method was able to distinguish thickness variation in the defect region. Moreover, the quantification of the defect’s thickness variation is more accurate with the multi-mode method. Therefore, theoretical and practical results prove that the variable-depth defect in metal plates can be successfully quantified and visualized by the proposed multi-mode electromagnetic ultrasonic LW CTI method. PMID:27144571

  3. The distribution of the Intermediate and Deep Water Masses on 60N in the Atlantic Ocean in 2006-2011.

    NASA Astrophysics Data System (ADS)

    Kolokolova, Alexandra

    2014-05-01

    Subpolar water structure in Atlantic Ocean has been analyzed using the newest hydrochemical data as well as CTD data. Water mass properties are compared to measurement data obtained in 2006, 2009 and 2011. New data analysis made it possible to reveal, that gradual Labrador Sea Water (LWS) dense lower core destruction in Irminger Basin and Icelandic Basin has been going on within the period of 2006-2011. In 2009 and 2011 dual-mode LWS structure was already not observed. New, developed Labrador Sea waters have spread in the upper layer. In addition to that arctic waters' influence increased slightly, and the tendency of intermediate southern-origin waters' intense penetration to the north continued. The detected LWS and Icelandic-Scotlandic Overflow Water (ISOW) warming, salinization and depletion in dissolved oxygen in Irminger Basin and Icelandic Basin have been going on in 2006-2011, happening simultaneously. Such a mode of water mass interaction in years 2000 is a reflection of the fact that one of the meridional circulation pattern dynamic phase is being completed. The mode is establishing, when water exchange between North Atlantic and Arctic Ocean is going to be defined by a weak current in the intermediate layer, intense entrance of deep waters from the north as well as strong subtropical Atlantic water transfer to the north.

  4. Regulation of photoreceptor gene expression by the retinal homeobox (Rx) gene product

    PubMed Central

    Pan, Yi; Martinez-De Luna, Reyna I.; Lou, Chih-Hong; Nekkalapudi, Srivamsi; Kelly, Lisa E.; Sater, Amy K.; El-Hodiri, Heithem M.

    2010-01-01

    The retinal homeobox (Rx) gene product is essential for eye development. However little is known about its molecular function. It has been demonstrated that Rx binds to photoreceptor conserved element (PCE-1), a highly conserved element found in the promoter region of photoreceptor-specific genes such as rhodopsin and red cone opsin. We verify that Rx is co-expressed with rhodopsin and red cone opsin in maturing photoreceptors and demonstrate that Rx binds to the rhodopsin and red cone opsin promoters in vivo. We also find that Rx can cooperate with the Xenopus analogs of Crx and Nrl, otx5b and XLMaf (respectively), to activate a Xenopus opsin promoter-dependent reporter. Finally, we demonstrate that reduction of Rx expression in tadpoles results in decreases in expression of several PCE-1 containing photoreceptor genes, abnormal photoreceptor morphology, and impaired vision. Our data suggests that Rx, in combination with other transcription factors, is necessary for normal photoreceptor gene expression, maintenance, and function. This establishes a direct role for Rx in regulation of genes expressed in a differentiated cell type. PMID:20060393

  5. Photoreceptor disc morphogenesis: The classical evagination model prevails.

    PubMed

    Pugh, Edward N

    2015-11-01

    Vision begins in photoreceptor outer segments with light captured by opsins in continually synthesized disc membranes. The process by which rod photoreceptor discs are formed has been controversial. In this issue, Ding et al. (2015. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201508093) show conclusively that rod discs are formed by plasma membrane evagination. PMID:26527745

  6. Visual responses in mice lacking critical components of all known retinal phototransduction cascades.

    PubMed

    Allen, Annette E; Cameron, Morven A; Brown, Timothy M; Vugler, Anthony A; Lucas, Robert J

    2010-01-01

    The mammalian visual system relies upon light detection by outer-retinal rod/cone photoreceptors and melanopsin-expressing retinal ganglion cells. Gnat1(-/-);Cnga3(-/-);Opn4(-/-) mice lack critical elements of each of these photoreceptive mechanisms via targeted disruption of genes encoding rod ? transducin (Gnat1); the cone-specific ?3 cyclic nucleotide gated channel subunit (Cnga3); and melanopsin (Opn4). Although assumed blind, we show here that these mice retain sufficiently widespread retinal photoreception to drive a reproducible flash electroretinogram (ERG). The threshold sensitivity of this ERG is similar to that of cone-based responses, however it is lost under light adapted conditions. Its spectral efficiency is consistent with that of rod opsin, but not cone opsins or melanopsin, indicating that it originates with light absorption by the rod pigment. The TKO light response survives intravitreal injection of U73122 (a phospholipase C antagonist), but is inhibited by a missense mutation of cone ? transducin (Gnat2(cpfl3)), suggesting Gnat2-dependence. Visual responses in TKO mice extend beyond the retina to encompass the lateral margins of the lateral geniculate nucleus and components of the visual cortex. Our data thus suggest that a Gnat1-independent phototransduction mechanism downstream of rod opsin can support relatively widespread responses in the mammalian visual system. This anomalous rod opsin-based vision should be considered in experiments relying upon Gnat1 knockout to silence rod phototransduction. PMID:21124780

  7. Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding.

    PubMed

    Vishnivetskiy, Sergey A; Ostermaier, Martin K; Singhal, Ankita; Panneels, Valerie; Homan, Kristoff T; Glukhova, Alisa; Sligar, Stephen G; Tesmer, John J G; Schertler, Gebhard F X; Standfuss, Joerg; Gurevich, Vsevolod V

    2013-11-01

    The effects of activating mutations associated with night blindness on the stoichiometry of rhodopsin interactions with G protein-coupled receptor kinase 1 (GRK1) and arrestin-1 have not been reported. Here we show that the monomeric form of WT rhodopsin and its constitutively active mutants M257Y, G90D, and T94I, reconstituted into HDL particles are effectively phosphorylated by GRK1, as well as two more ubiquitously expressed subtypes, GRK2 and GRK5. All versions of arrestin-1 tested (WT, pre-activated, and constitutively monomeric mutants) bind to monomeric rhodopsin and show the same selectivity for different functional forms of rhodopsin as in native disc membranes. Rhodopsin phosphorylation by GRK1 and GRK2 promotes arrestin-1 binding to a comparable extent, whereas similar phosphorylation by GRK5 is less effective, suggesting that not all phosphorylation sites on rhodopsin are equivalent in promoting arrestin-1 binding. The binding of WT arrestin-1 to phospho-opsin is comparable to the binding to its preferred target, P-Rh*, suggesting that in photoreceptors arrestin-1 only dissociates after opsin regeneration with 11-cis-retinal, which converts phospho-opsin into inactive phospho-rhodopsin that has lower affinity for arrestin-1. Reduced binding of arrestin-1 to the phospho-opsin form of G90D mutant likely contributes to night blindness caused by this mutation in humans. PMID:23872075

  8. Molecular genetics of rhodopsin and phototrans duction in the visual system of Drosophila

    SciTech Connect

    Zuker, C.; Cowman, A.; Montell, C.; Rubin, G.

    1987-05-01

    The authors have isolated the genes encoding four Drosophila visual pigments. Each of these opsins is expressed in a set of functionally and anatomically distinct photoreceptor cells of the eye. One is expressed in the six outer photoreceptor cells (R1-R6), the second in the central R8 photoreceptor cell, and the other two in the UV sensitive R7 photoreceptor cells. They have determined the structure and nucleotide sequence of each of these genes. They have used P element-mediated gene transfer to introduce the cloned structural gene for the R1-R6 opsin in the Drosophila germline and restored the ninaE mutant phenotype to wild-type. In an attempt to study the contribution of the various opsins to the specific functional properties of the different photoreceptor cell types, they have genetically engineered Drosophila lines that express R8 opsin in the R1-R6 photoreceptor cells. In collaboration with Drs. Ozaki and Pak at Purdue University, they have used oligonucleotide site-directed mutagenesis to mutate selected amino acids and regions of the rhodopsin molecule and reintroduced the mutated genes into Drosophila to analyze structure-function relationships in the rhodopsin molecule.

  9. Differential expression of photoreceptor-specific genes in the retina of a zebrafish cadherin2 mutant glass onion and zebrafish cadherin4 morphants.

    PubMed

    Liu, Q; Frey, R A; Babb-Clendenon, S G; Liu, B; Francl, J; Wilson, A L; Marrs, J A; Stenkamp, D L

    2007-01-01

    Cadherins are Ca2+ -dependent transmembrane molecules that mediate cell-cell adhesion through homophilic interactions. Cadherin2 (also called N-cadherin) and cadherin4 (also called R-cadherin), members of the classic cadherin subfamily, have been shown to be involved in development of a variety of tissues and organs including the visual system. To gain insight into cadherin2 and cadherin4 function in differentiation of zebrafish photoreceptors, we have analyzed expression patterns of several photoreceptor-specific genes (crx, gnat1, gnat2, irbp, otx5, rod opsin, rx1, and uv opsin) and/or a cone photoreceptor marker (zpr-1) in the retina of a zebrafish cadherin2 mutant, glass onion (glo) and in zebrafish embryos injected with a cadherin4 specific antisense morpholino oligonucleotide (cdh4MO). We find that expression of all these genes, and of zpr-1, is greatly reduced in the retina of both the glo and cadherin4 morphants. Moreover, in these embryos, expression of some genes (e.g. gnat1, gnat2 and irbp) is more affected than others (e.g. rod opsin and uv opsin). In embryos with both cadherins functions blocked (glo embryos injected with the cdh4MO), the eye initially formed, but became severely and progressively disintegrated and expressed little or no crx and otx5 as development proceeded. Our results suggest that cadherin2 and cadherin4 play important roles in the differentiation of zebrafish retinal photoreceptors. PMID:17070801

  10. Positive selection of a duplicated UV-sensitive visual pigment coincides with wing pigment evolution in Heliconius butterflies

    PubMed Central

    Briscoe, Adriana D.; Bybee, Seth M.; Bernard, Gary D.; Yuan, Furong; Sison-Mangus, Marilou P.; Reed, Robert D.; Warren, Andrew D.; Llorente-Bousquets, Jorge; Chiao, Chuan-Chin

    2010-01-01

    The butterfly Heliconius erato can see from the UV to the red part of the light spectrum with color vision proven from 440 to 640 nm. Its eye is known to contain three visual pigments, rhodopsins, produced by an 11-cis-3-hydroxyretinal chromophore together with long wavelength (LWRh), blue (BRh) and UV (UVRh1) opsins. We now find that H. erato has a second UV opsin mRNA (UVRh2)—a previously undescribed duplication of this gene among Lepidoptera. To investigate its evolutionary origin, we screened eye cDNAs from 14 butterfly species in the subfamily Heliconiinae and found both copies only among Heliconius. Phylogeny-based tests of selection indicate positive selection of UVRh2 following duplication, and some of the positively selected sites correspond to vertebrate visual pigment spectral tuning residues. Epi-microspectrophotometry reveals two UV-absorbing rhodopsins in the H. erato eye with λmax = 355 nm and 398 nm. Along with the additional UV opsin, Heliconius have also evolved 3-hydroxy-DL-kynurenine (3-OHK)-based yellow wing pigments not found in close relatives. Visual models of how butterflies perceive wing color variation indicate this has resulted in an expansion of the number of distinguishable yellow colors on Heliconius wings. Functional diversification of the UV-sensitive visual pigments may help explain why the yellow wing pigments of Heliconius are so colorful in the UV range compared to the yellow pigments of close relatives lacking the UV opsin duplicate. PMID:20133601

  11. Zebrafish melanopsin: isolation, tissue localisation and phylogenetic position.

    PubMed

    Bellingham, James; Whitmore, David; Philp, Alisdair R; Wells, Dominic J; Foster, Russell G

    2002-11-15

    Photoreception is best understood in retinal rods and cones, but it is not confined to these cells. In non-mammals, intrinsically photosensitive cells have been identified within several structures including the pineal, hypothalamus and skin. More recently novel light sensitive cells have been identified in the inner/basal retina of both teleosts and rodents. Melanopsin has been proposed as the photopigment mediating many of these non-rod, non-cone responses to light. However, much about the melanopsin gene family remains to be clarified including their potential role as photopigments, and taxonomic distribution. We have isolated the first orthologue of melanopsin from a teleost fish and show expression of this gene in a sub-set of retinal horizontal cells (type B). Zebrafish melanopsin, and orthologues of this gene, differ markedly from the vertebrate photopigment opsins. The putative counterion is not a glutamate but a tyrosine, the putative G-protein binding domain in the third cytoplasmic loop is not conserved, and they show low levels of amino acid identity (approximately 27%) to both the known photopigment opsins and to other members of the melanopsin family. Mouse melanopsin is only 58% identical to Xenopus, and 68% identical to zebrafish. By contrast, the photosensory opsin families show approximately 75% conservation. On the basis of their structure, genomic organisation, discrete evolutionary lineage, and their co-expression with other opsins, the melanopins are not obvious photosensory opsins. They might represent a separate branch of photopigment evolution in the vertebrates or they may have a non-direct photosensory function, perhaps as a photoisomerase, in non-rod, non-cone light detection. PMID:12487121

  12. The expression of retinal cell markers in human retinal pigment epithelial cells and their augmentation by the synthetic retinoid fenretinide

    PubMed Central

    Vugler, Anthony A.; Yu, Lu; Semo, Maayan; Coffey, Pete; Moss, Stephen E.; Greenwood, John

    2011-01-01

    Purpose In several species the retinal pigment epithelium (RPE) has the potential to transdifferentiate into retinal cells to regenerate functional retinal tissue after injury. However, this capacity for regeneration is lost in mammals. The synthetic retinoic acid derivative, fenretinide [N(4-hydroxyphenyl) retinamide], induces a neuronal-like phenotype in the human adult retinal pigment epithelial cell line (ARPE-19). These changes are characterized by the appearance of neural-like processes and the expression of neuronal markers not normally associated with RPE cells. Here we assess whether fenretinide can induce a neuroretinal cell phenotype in ARPE-19 cells, by examining retinal cell marker expression. Methods ARPE-19 cells were treated daily with culture medium containing either 3 μM fenretinide or dimethyl sulfoxide as a control for 7 days. Cells were processed for immunocytochemistry, western blotting, and for analysis by PCR to examine the expression of a panel of RPE, neural, and retinal-associated cellular markers, including classical and non-canonical opsins. Results Treatment with fenretinide for 7 days induced the formation of neuronal-like processes in ARPE-19 cells. Fenretinide induced the expression of the cone long wavelength sensitive opsin (OPN1lw) but not rhodopsin (RHO), while decreasing the expression of RPE cell markers. Many of the neuronal and retinal specific markers examined were expressed in both control and fenretinide treated cells, including those involved in photoreceptor cell development and the multipotency of neural retinal progenitor cells. Interestingly, ARPE-19 cells also expressed both photoreceptor specific and non-specific canonical opsins. Conclusions The expression of retinal-associated markers and loss of RPE cell markers in control ARPE-19 cells suggests that these cells might have dedifferentiated from an RPE cell phenotype under standard culture conditions. The expression of molecules, such as the transcription factors paired box 6 gene (PAX6), sex determining region Y-box 2 (SOX2), cone-rod homeobox (CRX), and neural retina leucine zipper (NRL), further implies that in culture these cells are predisposed toward a retinal progenitor-like state. The fenretinide-induced increase in photoreceptor cell markers, accompanied by a decrease in RPE cell markers, suggests that retinoids may play a role in the transdifferentiation of RPE cells. Importantly, our data show for the first time the expression of a vertebrate ciliary opsin (OPN1lw) and rhabdomeric-like opsin, opsin 4 (OPN4 also known as melanopsin) in a clonal cell line. Together these data suggest that ARPE-19 cells are primed for and possess the capacity to differentiate toward a retinal cell-like lineage. PMID:21738400

  13. Breeding objectives for pigs in Kenya. II: economic values incorporating risks in different smallholder production systems.

    PubMed

    Mbuthia, Jackson Mwenda; Rewe, Thomas Odiwuor; Kahi, Alexander Kigunzu

    2015-02-01

    This study estimated economic values for production traits (dressing percentage (DP), %; live weight for growers (LWg), kg; live weight for sows (LWs), kg) and functional traits (feed intake for growers (FEEDg), feed intake for sow (FEEDs), preweaning survival rate (PrSR), %; postweaning survival (PoSR), %; sow survival rate (SoSR), %, total number of piglets born (TNB) and farrowing interval (FI), days) under different smallholder pig production systems in Kenya. Economic values were estimated considering two production circumstances: fixed-herd and fixed-feed. Under the fixed-herd scenario, economic values were estimated assuming a situation where the herd cannot be increased due to other constraints apart from feed resources. The fixed-feed input scenario assumed that the herd size is restricted by limitation of feed resources available. In addition to the tradition profit model, a risk-rated bio-economic model was used to derive risk-rated economic values. This model accounted for imperfect knowledge concerning risk attitude of farmers and variance of input and output prices. Positive economic values obtained for traits DP, LWg, LWs, PoSR, PrSR, SoSR and TNB indicate that targeting them in improvement would positively impact profitability in pig breeding programmes. Under the fixed-feed basis, the risk-rated economic values for DP, LWg, LWs and SoSR were similar to those obtained under the fixed-herd situation. Accounting for risks in the EVs did not yield errors greater than ±50 % in all the production systems and basis of evaluation meaning there would be relatively little effect on the real genetic gain of a selection index. Therefore, both traditional and risk-rated models can be satisfactorily used to predict profitability in pig breeding programmes. PMID:25433647

  14. Impedance-matching analysis in IR leaky-wave antennas

    NASA Astrophysics Data System (ADS)

    Premkumar, Navaneeth; Xu, Yuancheng; Lail, Brian A.

    2015-08-01

    Planar leaky-wave antennas (LWA) that are capable of full-space scanning have long since been the pursuit for applications including, but not limited to, integration onto vehicles and into cameras for wide-angle of view beam-steering. Such a leaky-wave surface (LWS) was designed for long-wave infrared frequencies with frequency scanning capability. The LWS is based on a microstrip patch array design of a leaky-wave impedance surface and is made up of gold microstrip patches on a grounded zinc sulphide substrate. A 1D composite right/left-handed (CRLH) metamaterial made by periodically stacking a unit cell of the LWS in the longitudinal direction to form a LWA was designed. This paper deals with loading the LWA with a nickel bolometer to collect leaky-wave signals. The LWA radiates a backward leaking wave at 30 degrees at 28.3THz and scans through broadside for frequencies 20THz through 40THz. The paper deals with effectively placing the bolometer in order for the collected signal to exhibit the designed frequency regime. An effective way to maximize the power coupling into the load from the antenna is also explored. The benefit of such a metamaterial/holographic antennacoupled detector is its ability to provide appreciable capture cross-sections while delivering smart signals to subwavelength sized detectors. Due to their high-gain, low-profile, fast response time of the detector and ease of fabrication, this IR LWA-coupled bolometer harbors great potential in the areas of high resolution, uncooled, infrared imaging.

  15. WP1: transgenic opto-animals

    NASA Astrophysics Data System (ADS)

    UŻarowska, E.; Czajkowski, Rafał; Konopka, W.

    2014-11-01

    We aim to create a set of genetic tools where permanent opsin expression (ChR or NpHR) is precisely limited to the population of neurons that express immediate early gene c-fos during a specific temporal window of behavioral training. Since the c-fos gene is only expressed in neurons that form experience-dependent ensemble, this approach will result in specific labeling of a small subset of cells that create memory trace for the learned behavior. To this end we employ two alternative inducible gene expression systems: Tet Expression System and Cre/lox System. In both cases, the temporal window for opsin induction is controlled pharmacologically, by doxycycline or tamoxifen, respectively. Both systems will be used for creating lines of transgenic animals.

  16. Colour vision in marine organisms.

    PubMed

    Marshall, Justin; Carleton, Karen L; Cronin, Thomas

    2015-10-01

    Colour vision in the marine environment is on average simpler than in terrestrial environments with simple or no colour vision through monochromacy or dichromacy. Monochromacy is found in marine mammals and elasmobranchs, including whales and sharks, but not some rays. Conversely, there is also a greater diversity of colour vision in the ocean than on land, examples being the polyspectral stomatopods and the many colour vision solutions found among reef fish. Recent advances in sequencing reveal more opsin (visual pigment) types than functionally useful at any one time. This diversity arises through opsin duplication and conversion. Such mechanisms allow pick-and-mix adaptation that tunes colour vision on a variety of very short non-evolutionary timescales. At least some of the diversity in marine colour vision is best explained as unconventional colour vision or as neutral drift. PMID:25725325

  17. UV wavelengths experienced during development affect larval newt visual sensitivity and predation efficiency.

    PubMed

    Martin, Mélissa; Théry, Marc; Rodgers, Gwendolen; Goven, Delphine; Sourice, Stéphane; Mège, Pascal; Secondi, Jean

    2016-02-01

    We experimentally investigated the influence of developmental plasticity of ultraviolet (UV) visual sensitivity on predation efficiency of the larval smooth newt, Lissotriton vulgaris. We quantified expression of SWS1 opsin gene (UV-sensitive protein of photoreceptor cells) in the retinas of individuals who had developed in the presence (UV+) or absence (UV-) of UV light (developmental treatments), and tested their predation efficiency under UV+ and UV- light (testing treatments). We found that both SWS1 opsin expression and predation efficiency were significantly reduced in the UV- developmental group. Larvae in the UV- testing environment displayed consistently lower predation efficiency regardless of their developmental treatment. These results prove for the first time, we believe, functional UV vision and developmental plasticity of UV sensitivity in an amphibian at the larval stage. They also demonstrate that UV wavelengths enhance predation efficiency and suggest that the magnitude of the behavioural response depends on retinal properties induced by the developmental lighting environment. PMID:26843556

  18. Dark adaptaion processes in the amphibian rod.

    PubMed

    Donner, K O

    1977-06-29

    Rod dark adaptation in the amphibian retina appears to be due to three processes: 1. background adaptation, occurring immediately after the extinction of an adapting or bleaching light, 2. intermediate adaptation, that frequently lasts 30 min or more and 3. opsin adaptation, which in the isolated retina where regeneration of rhodopsin is insignificant, is observed a a permanent loss of sensitivity after the completion of intermediate adaptation. Intermediate adaptation is characterized by a linear relation between log threshold and the amount of "retinal" present, a similar relation is obtained between log threshold and the amount of rhodopsin bleached in opsin adaptation. These adaptation processes are discussed in terms of a model of the rod outer segment. PMID:890054

  19. Connecting the navigational clock to sun compass input in monarch butterfly brain.

    PubMed

    Sauman, Ivo; Briscoe, Adriana D; Zhu, Haisun; Shi, Dingding; Froy, Oren; Stalleicken, Julia; Yuan, Quan; Casselman, Amy; Reppert, Steven M

    2005-05-01

    Migratory monarch butterflies (Danaus plexippus) use a time-compensated sun compass to navigate to their overwintering grounds in Mexico. Although polarized light is one of the celestial cues used for orientation, the spectral content (color) of that light has not been fully explored. We cloned the cDNAs of three visual pigment-encoding opsins (ultraviolet [UV], blue, and long wavelength) and found that all three are expressed uniformly in main retina. The photoreceptors of the polarization-specialized dorsal rim area, on the other hand, are monochromatic for the UV opsin. Behavioral studies support the importance of polarized UV light for flight orientation. Next, we used clock protein expression patterns to identify the location of a circadian clock in the dorsolateral protocerebrum of butterfly brain. To provide a link between the clock and the sun compass, we identified a CRYPTOCHROME-staining neural pathway that likely connects the circadian clock to polarized light input entering brain. PMID:15882645

  20. Inhibiting the Activity of CA1 Hippocampal Neurons Prevents the Recall of Contextual Fear Memory in Inducible ArchT Transgenic Mice

    PubMed Central

    Sakaguchi, Masanori; Kim, Karam; Yu, Lily Mae Yee; Hashikawa, Yoshiko; Sekine, Yukiko; Okumura, Yuki; Kawano, Masako; Hayashi, Masanobu; Kumar, Deependra; Boyden, Edward S.; McHugh, Thomas J.; Hayashi, Yasunori

    2015-01-01

    The optogenetic manipulation of light-activated ion-channels/pumps (i.e., opsins) can reversibly activate or suppress neuronal activity with precise temporal control. Therefore, optogenetic techniques hold great potential to establish causal relationships between specific neuronal circuits and their function in freely moving animals. Due to the critical role of the hippocampal CA1 region in memory function, we explored the possibility of targeting an inhibitory opsin, ArchT, to CA1 pyramidal neurons in mice. We established a transgenic mouse line in which tetracycline trans-activator induces ArchT expression. By crossing this line with a CaMKIIα-tTA transgenic line, the delivery of light via an implanted optrode inhibits the activity of excitatory CA1 neurons. We found that light delivery to the hippocampus inhibited the recall of a contextual fear memory. Our results demonstrate that this optogenetic mouse line can be used to investigate the neuronal circuits underlying behavior. PMID:26075894

  1. Neuropsin (OPN5)-mediated photoentrainment of local circadian oscillators in mammalian retina and cornea

    PubMed Central

    Buhr, Ethan D.; Yue, Wendy W. S.; Ren, Xiaozhi; Jiang, Zheng; Liao, Hsi-Wen Rock; Mei, Xue; Vemaraju, Shruti; Nguyen, Minh-Thanh; Reed, Randall R.; Lang, Richard A.; Yau, King-Wai; Van Gelder, Russell N.

    2015-01-01

    The molecular circadian clocks in the mammalian retina are locally synchronized by environmental light cycles independent of the suprachiasmatic nuclei (SCN) in the brain. Unexpectedly, this entrainment does not require rods, cones, or melanopsin (OPN4), possibly suggesting the involvement of another retinal photopigment. Here, we show that the ex vivo mouse retinal rhythm is most sensitive to short-wavelength light but that this photoentrainment requires neither the short-wavelength–sensitive cone pigment [S-pigment or cone opsin (OPN1SW)] nor encephalopsin (OPN3). However, retinas lacking neuropsin (OPN5) fail to photoentrain, even though other visual functions appear largely normal. Initial evidence suggests that OPN5 is expressed in select retinal ganglion cells. Remarkably, the mouse corneal circadian rhythm is also photoentrainable ex vivo, and this photoentrainment likewise requires OPN5. Our findings reveal a light-sensing function for mammalian OPN5, until now an orphan opsin. PMID:26392540

  2. WASTE CERTIFICATION PROGRAM PLAN - REVISION 7

    SciTech Connect

    MORGAN, LK

    2002-01-08

    The primary changes that have been made to this revision reflect the relocation of the Waste Certification Official (WCO) organizationally from the Quality Services Division (QSD) into the Laboratory Waste Services (LWS) Organization. Additionally, the responsibilities for program oversight have been differentiated between the QSD and LWS. The intent of this effort is to ensure that those oversight functions, which properly belonged to the WCO, moved with that function; but retain an independent oversight function outside of the LWS Organization ensuring the potential for introduction of organizational bias, regarding programmatic and technical issues, is minimized. The Waste Certification Program (WCP) itself has been modified to allow the waste certification function to be performed by any of the personnel within the LWS Waste Acceptance/Certification functional area. However, a single individual may not perform both the technical waste acceptance review and the final certification review on the same 2109 data package. Those reviews must be performed by separate individuals in a peer review process. There will continue to be a designated WCO who will have lead programmatic responsibility for the WCP and will exercise overall program operational oversite as well as determine the overall requirements of the certification program. The quality assurance organization will perform independent, outside oversight to ensure that any organizational bias does not degrade the integrity of the waste certification process. The core elements of the previous WCP have been retained, however, the terms and process structure have been modified.. There are now two ''control points,'' (1) the data package enters the waste certification process with the signature of the Generator Interface/Generator Interface Equivalent (GI/GIE), (2) the package is ''certified'', thus exiting the process. The WCP contains three steps, (1) the technical review for waste acceptance, (2) a review of the packaging and labeling (the old Control Point 3), and (3) programmatic review and certification. The signature of the individual performing the certification review resulting in certification of the package constitutes Control Point 2 in the revised process.

  3. Space Weather Monitoring with DSCOVR and Beyond

    NASA Astrophysics Data System (ADS)

    Szabo, A.

    2013-05-01

    With the NOAA/NASA DSCOVR spacecraft launched around 2015, continued near-Earth solar wind monitoring will be assured. However, to provide more than the current 30-45 minutes warning time of incoming solar transients, newer platforms are required. Solar sail missions would orbit an artificial 1st Lagrange point nearly doubling the current warning time. Four or five LWS Solar Sentinels orbiting the Sun inside 0.5 AU would provide warning times close to 1 day. The required scientific understanding is already at hand to take full advantage of such new missions. DSCOVR and these new missions will be discussed in detail.

  4. Using Digital Globes to Explore the Deep Sea and Advance Public Literacy in Earth System Science

    NASA Astrophysics Data System (ADS)

    Beaulieu, S. E.; Brickley, A.; Emery, M.; Spargo, A.; Patterson, K.; Joyce, K.; Silva, T.; Madin, K.

    2014-12-01

    Digital globes are new technologies increasingly used in both informal and formal education to display global datasets. By creating a narrative using multiple datasets, linkages between Earth systems - lithosphere, hydrosphere, atmosphere, and biosphere - can be conveyed. But how effective are digital globes in advancing public literacy in Earth system science? We addressed this question in developing new content for digital globes that interweaves imagery obtained by deep-diving vehicles with global datasets, including a new dataset locating the world's known hydrothermal vents. Our two narratives, "Life Without Sunlight" (LWS) and "Smoke and Fire Underwater" (SFU), each focus on STEM (science, technology, engineering, and mathematics) principles related to geology, biology, and exploration. We are preparing a summative evaluation for our content delivered on NOAA's Science on a Sphere as interactive presentations and as movies. We tested knowledge gained with respect to the STEM principles and the level of excitement generated by the virtual deep-sea exploration. We conducted a Post-test Only Design with quantitative data based on self-reporting on a Likert scale. A total of 75 adults and 48 youths responded to our questionnaire, distributed into test groups that saw either one of the two narratives delivered either as a movie or as an interactive presentation. Here, we report preliminary results for the youths, the majority (81%) of which live in towns with lower income and lower levels of educational attainment as compared to other towns in Massachusetts. For both narratives, there was knowledge gained for all 6 STEM principles and "Quite a Bit" of excitement. The mode in responses for knowledge gained was "Quite a Bit" for both the movie and the interactive presentation for 4 of the STEM principles (LWS geology, LWS biology, SFU geology, and SFU exploration) and "Some" for SFU biology. Only for LWS exploration was there a difference in mode between the interactive presentation ("A Little") and the movie ("Quite a Bit"). We conclude that our content for digital globes is effective in teaching the STEM principles and exciting viewers about the deep ocean frontier. We attribute this success to the tight collaboration between scientists, educators, and graphic artists in developing the content for public audiences.

  5. VizieR Online Data Catalog: VY CMa molecular line spectra (Matsuura+, 2014)

    NASA Astrophysics Data System (ADS)

    Matsuura, M.; Yates, J. A.; Barlow, M. J.; Swinyard, B. M.; Royer, P.; Cernicharo, J.; Decin, L.; Wesson, R.; Polehampton, E. T.; Blommaert, J. A. D. L.; Groenewegen, M. A. T.; van de Steene, G. C.; van Hoof, P. A. M.

    2015-03-01

    The Herschel Space Observatory (hereafter Herschel) was launched in 2009 May with three instruments on board: SPIRE, PACS and HIFI . We report here observations made with the SPIRE Fourier Transform Spectrometer (FTS) and with the PACS grating spectrometer, which together cover the wavelength range from 55 to 650um. The SPIRE FTS covers the 190-650um wavelength range, simultaneously, while the PACS spectrometer covers the 55-210um spectral range, similar to that covered by the Long Wavelength Spectrometer (LWS) on board the Infrared Space Observatory (ISO), although with higher spectral resolution and greater sensitivity. (2 data files).

  6. Thermal Evolution of Juvenile Subduction Zones ' New Constraints from Lu-Hf Geochronology on HP oceanic rocks (Halilbaǧi, Central Anatolia)

    NASA Astrophysics Data System (ADS)

    Pourteau, Amaury; Scherer, Erik; Schmidt, Alexander; Bast, Rebecca

    2015-04-01

    The thermal structure of subduction zones plays a key role on mechanical and chemical processes taking place along the slab-mantle interface. Until now, changes through time of this thermal structure have been explored mostly by the means of numerical simulations. However, both "warm" (i.e., epidote-bearing), and "cold" (i.e., lawsonite-bearing) HP oceanic rocks have been reported in some fossil subduction complexes exposed at the Earth's surface (e.g., Franciscan Complex, California; Rio San Juan Complex, Hispañola; Halilbağı Unit, Central Anatolia). These a-priori "incompatible" rocks witness different thermal stages of ancient subduction zones and their study might provide complementary constraints to numerical models. To decipher the meaning of these contrasting metamorphic rocks in the Halilbağı Unit, we are carrying out Lu-Hf geochronology on garnet (grt) and lws from a variety of HP oceanic rocks, as well as the metamorphic sole of the overlying ophiolite. We selected five samples that are representative of the variety of metamorphic evolutions (i.e. peak conditions and P-T paths) encountered in this area. Preliminary analyses yielded 110 Ma (grt-hbl isochron) for a sub-ophiolitic grt amphibolite; 92 Ma (grt-omp) for an eclogite with prograde and retrograde ep; 90 Ma (grt-omp) for an eclogitic metabasite with prograde ep and retrograde ep+lws; 87 Ma (grt-gln) for a lws eclogite with prograde ep; and 86 Ma (grt-gln) for a blueschist with prograde and retrograde lws. These ages are mainly two-point isochrons. Further-refined data will be presented at the EGU General Assembly 2015, in Vienna. The consistent younging trend from "warm" to "cold" metamorphic rocks revealed by these first-order results points to metamorphic-sole formation during the initiation of intra-oceanic subduction at ~110 Ma, and subsequent cooling of the slab-mantle interface between 92 and 86 Ma. Therefore, the contrasting metamorphic evolutions encountered in the Halilbağı Unit record the progressive thermal maturation of the juvenile Neotethyan subduction zone. This period of ~23 myr between subduction initiation and thermal "steady state" is significantly shorter than that obtained for the Rio San Juan Complex (~60 myr; Krebs et al. 2008, Lithos, 103, 106-137), but compares well with that for the Franciscan Complex (~22 myr; Anczkiewicz et al. 2004, EPSL, 225, 147-161) and falls in the range predicted in numerical simulations (e.g., Gerya et al. 2002, Tectonics, 21/6, 1056).

  7. The Living With a Star Space Environment Testbed Program

    NASA Technical Reports Server (NTRS)

    Barth, Janet; LaBel, Kenneth; Day, John H. (Technical Monitor)

    2001-01-01

    NASA has initiated the Living with a Star (LWS) Program to develop the scientific understanding to address the aspects of the Connected Sun-Earth system that affects life and society. The Program Architecture includes science missions, theory and modeling and Space Environment Testbeds (SET). This current paper discusses the Space Environment Testbeds. The goal of the SET program is to improve the engineering approach to accomodate and/or mitigate the effects of solar variability on spacecraft design and operations. The SET Program will infuse new technologies into the space programs through collection of data in space and subsequent design and validation of technologies. Examples of these technologies are cited and discussed.

  8. The Living With a Star Space Environment Testbed Experiments

    NASA Technical Reports Server (NTRS)

    Xapsos, Michael A.

    2014-01-01

    The focus of the Living With a Star (LWS) Space Environment Testbed (SET) program is to improve the performance of hardware in the space radiation environment. The program has developed a payload for the Air Force Research Laboratory (AFRL) Demonstration and Science Experiments (DSX) spacecraft that is scheduled for launch in August 2015 on the SpaceX Falcon Heavy rocket. The primary structure of DSX is an Evolved Expendable Launch Vehicle (EELV) Secondary Payload Adapter (ESPA) ring. DSX will be in a Medium Earth Orbit (MEO). This oral presentation will describe the SET payload.

  9. Designing 2D arrays for SHM of planar structures: a review

    NASA Astrophysics Data System (ADS)

    Stepinski, Tadeusz; Ambrozinski, Lukasz; Uhl, Tadeusz

    2013-04-01

    Monitoring structural integrity of large planar structures that aims at detecting and localizing impact or damage at any point of the structure requires normally a relatively dense network of uniformly distributed ultrasonic sensors. 2-D ultrasonic phased arrays, due to their beam-steering capability and all azimuth angle coverage are a very promising tool for structural health monitoring (SHM) of plate-like structures using Lamb waves (LW). Linear phased arrays that have been proposed for that purpose, produce mirrored image characterized by azimuth dependent resolution, which prevents unequivocal damage localization. 2D arrays do not have this drawback and they are even capable of mode selectivity when generating and receiving LWs. Performance of 2D arrays depends on their topology as well as the number of elements (transducers) used and their spacing in terms of wavelength. In this paper we propose a consistent methodology for three-step: theoretical, numerical and experimental investigation of a diversity of 2D array topologies in SHM applications. In the first step, the theoretical evaluation is performed using frequency-dependent structure transfer function (STF). STF that defines linear propagation of different LWs modes through the dispersive medium enables theoretical investigation of the particular array performance for a predefined tone-burst excitation signal. A dedicated software tool has been developed for the numerical evaluation of 2D array directional characteristics (beampattern) in a specific structure. The simulations are performed using local interaction simulation approach (LISA), implemented using NVIDIA CUDA graphical computation unit (GPU), which enables time-efficient 3D simulations of LWs propagation. Beampatterns of a 2D array can be to some extend evaluated analytically and using numerical simulations; in most cases, however, they require experimental verification. Using scanning laser vibrometer is proposed for that purpose, in a setup where LWs, excited by PZT transmitters of the investigated array are sensed in multiple points corresponding to the locations of the 2D array receiving elements. A virtual receiving sub-array is created in this way and the performance of various array architectures in the reception mode can be evaluated experimentally without the need of physical prototype; a change of topology requires only straightforward modification of the measurement points distribution at the tested structure. For illustration, beampatterns of three symmetrical 2D topologies, i.e., circular, star-shaped and spiralshaped, will be examined in the paper and compared in terms of their beam-width and side-lobes level. The effect of apodization applied to the array elements will be also investigated.

  10. Common Transcriptional Mechanisms for Visual Photoreceptor Cell Differentiation among Pancrustaceans

    PubMed Central

    Mahato, Simpla; Morita, Shinichi; Tucker, Abraham E.; Liang, Xulong; Jackowska, Magdalena; Friedrich, Markus; Shiga, Yasuhiro; Zelhof, Andrew C.

    2014-01-01

    A hallmark of visual rhabdomeric photoreceptors is the expression of a rhabdomeric opsin and uniquely associated phototransduction molecules, which are incorporated into a specialized expanded apical membrane, the rhabdomere. Given the extensive utilization of rhabdomeric photoreceptors in the eyes of protostomes, here we address whether a common transcriptional mechanism exists for the differentiation of rhabdomeric photoreceptors. In Drosophila, the transcription factors Pph13 and Orthodenticle (Otd) direct both aspects of differentiation: rhabdomeric opsin transcription and rhabdomere morphogenesis. We demonstrate that the orthologs of both proteins are expressed in the visual systems of the distantly related arthropod species Tribolium castaneum and Daphnia magna and that their functional roles are similar in these species. In particular, we establish that the Pph13 homologs have the ability to bind a subset of Rhodopsin core sequence I sites and that these sites are present in key phototransduction genes of both Tribolium and Daphnia. Furthermore, Pph13 and Otd orthologs are capable of executing deeply conserved functions of photoreceptor differentiation as evidenced by the ability to rescue their respective Drosophila mutant phenotypes. Pph13 homologs are equivalent in their ability to direct both rhabdomere morphogenesis and opsin expression within Drosophila, whereas Otd paralogs demonstrate differential abilities to regulate photoreceptor differentiation. Finally, loss-of-function analyses in Tribolium confirm the conserved requirement of Pph13 and Otd in regulating both rhabdomeric opsin transcription and rhabdomere morphogenesis. Taken together, our data identify components of a regulatory framework for rhabdomeric photoreceptor differentiation in Pancrustaceans, providing a foundation for defining ancestral regulatory modules of rhabdomeric photoreceptor differentiation. PMID:24991928

  11. Molecular ecology and adaptation of visual photopigments in craniates.

    PubMed

    Davies, Wayne I L; Collin, Shaun P; Hunt, David M

    2012-07-01

    In craniates, opsin-based photopigments expressed in the eye encode molecular 'light sensors' that constitute the initial protein in photoreception and the activation of the phototransduction cascade. Since the cloning and sequencing of the first vertebrate opsin gene (bovine rod opsin) nearly 30 years ago (Ovchinnikov Yu 1982, FEBS Letters, 148, 179-191; Hargrave et al. 1983, Biophysics of Structure & Mechanism, 9, 235-244; Nathans & Hogness 1983, Cell, 34, 807-814), it is now well established that variation in the subtypes and spectral properties of the visual pigments that mediate colour and dim-light vision is a prevalent mechanism for the molecular adaptation to diverse light environments. In this review, we discuss the origins and spectral tuning of photopigments that first arose in the agnathans to sample light within the ancient aquatic landscape of the Early Cambrian, detailing the molecular changes that subsequently occurred in each of the opsin classes independently within the main branches of extant jawed gnathostomes. Specifically, we discuss the adaptive changes that have occurred in the photoreceptors of craniates as they met the ecological challenges to survive in quite differing photic niches, including brightly lit aquatic surroundings; the deep sea; the transition to and from land; diurnal, crepuscular and nocturnal environments; and light-restricted fossorial settings. The review ends with a discussion of the limitations inherent to the 'nocturnal-bottleneck' hypothesis relevant to the evolution of the mammalian visual system and a proposition that transition through a 'mesopic-bottleneck' may be a more appropriate model. PMID:22650357

  12. Deep tissue targeted near-infrared optogenetic stimulation using fully implantable upconverting light bulbs.

    PubMed

    Chamanzar, Maysamreza; Garfield, David J; Iafrati, Jillian; Sohal, Vikaas; Chan, Emory; Cohen, Bruce; Schuck, P James; Maharbiz, Michel M

    2015-08-01

    We demonstrate for the first time, the possibility of targeted optogenetic stimulation of neurons deep into brain tissue (>2 mm) in a minimally-invasive way by sending near-infrared light through tissue to excite passive lanthanide-doped blue-emitting upconverting nanocrystals (UCNPs) encapsulated in Parylene C microstructure light bulbs that emit visible (blue) light and locally excite opsins with high spatial resolution. PMID:26736388

  13. A High-Throughput Drug Screening Strategy for Detecting Rhodopsin P23H Mutant Rescue and Degradation

    PubMed Central

    Chen, Yuanyuan; Tang, Hong; Seibel, William; Papoian, Ruben; Li, Xiaoyu; Lambert, Nevin A.; Palczewski, Krzysztof

    2015-01-01

    Purpose. Inherent instability of the P23H mutant opsin accounts for approximately 10% of autosomal dominant retinitis pigmentosa cases. Our purpose was to develop an overall set of reliable screening strategies to assess if either stabilization or enhanced degradation of mutant rhodopsin could rescue rod photoreceptors expressing this mutant protein. These strategies promise to reveal active compounds and clarify molecular mechanisms of biologically important processes, such as inhibition of target degradation or enhanced target folding. Methods. Cell-based bioluminescence reporter assays were developed and validated for high-throughput screening (HTS) of compounds that promote either stabilization or degradation of P23H mutant opsin. Such assays were further complemented by immunoblotting and image-based analyses. Results. Two stabilization assays of P23H mutant opsin were developed and validated, one based on β-galactosidase complementarity and a second assay involving bioluminescence resonance energy transfer (BRET) technology. Moreover, two additional assays evaluating mutant protein degradation also were employed, one based on the disappearance of luminescence and another employing the ALPHA immunoassay. Imaging of cells revealed the cellular localization of mutant rhodopsin, whereas immunoblots identified changes in the aggregation and glycosylation of P23H mutant opsin. Conclusions. Our findings indicate that these initial HTS and following assays can identify active therapeutic compounds, even for difficult targets such as mutant rhodopsin. The assays are readily scalable and their function has been proven with model compounds. High-throughput screening, supported by automated imaging and classic immunoassays, can further characterize multiple steps and pathways in the biosynthesis and degradation of this essential visual system protein. PMID:25783607

  14. Colour tuning mechanisms of visual pigments.

    PubMed

    Lin, S W; Sakmar, T P

    1999-01-01

    Spectral tuning by visual pigments involves modulation of physical properties of the 11-cis-retinylidene protonated Schiff base (PSB) chromophore by amino acid side chains in and around the chromophore-binding pocket. Specific molecular contacts between the chromophore and the amino acid side chains of the opsin chromophore-binding pocket have been determined recently using an interdisciplinary approach consisting of site-directed mutagenesis, optical and vibrational spectroscopy, and molecular graphics modelling. These studies provide insight into the mechanism of spectral tuning among visual pigments. In blue pigments a majority of the opsin shift is caused by polar amino acid side chains arrayed about the PSB to increase the energy gap between the ground (S0) and excited states (S1). In addition, a specific tyrosine near the chromophore ring causes a decrease in solvent polarizability. Other amino acid residues alter the binding pocket structure to strengthen electrostatic interaction between the PSB and its counterion and/or solvent dipoles. In the green and red pigments, the work of Kochendoerfer et al (1997; Biochemistry 26:6577-6587) demonstrates that local structural perturbations at the PSB or elsewhere are not responsible for spectral tuning. Instead, the green-to-red opsin shift is best explained by dipolar side chains near the chromophore ring that lower the transition energy that occurs upon electronic excitation by affecting the change in electric dipole moment. In summary, the absorption maximum of a visual pigment is primarily regulated by the interaction of the chromophore charge distribution with dipolar residues in its opsin chromophore-binding pocket. The work presented in this paper is reported in greater detail in Lin et al. PMID:10614049

  15. The Giant Mottled Eel, Anguilla marmorata, Uses Blue-Shifted Rod Photoreceptors during Upstream Migration

    PubMed Central

    Wang, Feng-Yu; Fu, Wen-Chun; Wang, I-Li

    2014-01-01

    Catadromous fishes migrate between ocean and freshwater during particular phases of their life cycle. The dramatic environmental changes shape their physiological features, e.g. visual sensitivity, olfactory ability, and salinity tolerance. Anguilla marmorata, a catadromous eel, migrates upstream on dark nights, following the lunar cycle. Such behavior may be correlated with ontogenetic changes in sensory systems. Therefore, this study was designed to identify changes in spectral sensitivity and opsin gene expression of A. marmorata during upstream migration. Microspectrophotometry analysis revealed that the tropical eel possesses a duplex retina with rod and cone photoreceptors. The λmax of rod cells are 493, 489, and 489 nm in glass, yellow, and wild eels, while those of cone cells are 508, and 517 nm in yellow, and wild eels, respectively. Unlike European and American eels, Asian eels exhibited a blue-shifted pattern of rod photoreceptors during upstream migration. Quantitative gene expression analyses of four cloned opsin genes (Rh1f, Rh1d, Rh2, and SWS2) revealed that Rh1f expression is dominant at all three stages, while Rh1d is expressed only in older yellow eel. Furthermore, sequence comparison and protein modeling studies implied that a blue shift in Rh1d opsin may be induced by two known (N83, S292) and four putative (S124, V189, V286, I290) tuning sites adjacent to the retinal binding sites. Finally, expression of blue-shifted Rh1d opsin resulted in a spectral shift in rod photoreceptors. Our observations indicate that the giant mottled eel is color-blind, and its blue-shifted scotopic vision may influence its upstream migration behavior and habitat choice. PMID:25101636

  16. [Inherited colour vision deficiencies--from Dalton to molecular genetics].

    PubMed

    Cvetkovi?, Dragana; Cvetkovi?, Dobrosav

    2005-01-01

    In recent years, great advances have been made in our understanding of the molecular basis of colour vision defects, as well as of the patterns of genetic variation in individuals with normal colour vision. Molecular genetic analyses have explained the diversity of types and degrees of severity in colour vision anomalies, their frequencies, pronounced individual variations in test results, etc. New techniques have even enabled the determination of John Dalton's real colour vision defect, 150 years after his death. Inherited colour vision deficiencies most often result from the mutations of genes that encode cone opsins. Cone opsin genes are linked to chromosomes 7 (the S or "blue" gene) and X (the L or "red" gene and the M or "green" gene). The L and M genes are located on the q arm of the X chromosome in a head-to-tail array, composed of 2 to 6 (typically 3) genes--a single L is followed by one or more M genes. Only the first two genes of the array are expressed and contribute to the colour vision phenotype. The high degree of homology (96%) between the L and M genes predisposes them to unequal recombination, leading to gene deletion or the formation of hybrid genes (comprising portions of both the L and M genes), explaining the majority of the common red-green colour vision deficiencies. The severity of any deficiency is influenced by the difference in spectral sensitivity between the opsins encoded by the first two genes of the array. A rare defect, S monochromacy, is caused either by the deletion of the regulatory region of the array or by mutations that inactivate the L and M genes. Most recent research concerns the molecular basis of complete achromatopsia, a rare disorder that involves the complete loss of all cone function. This is not caused by mutations in opsin genes, but in other genes that encode cone-specific proteins, e.g. channel proteins and transducin. PMID:16758855

  17. Origin of the vertebrate visual cycle: III. Distinct distribution of RPE65 and beta-carotene 15,15'-monooxygenase homologues in Ciona intestinalis.

    PubMed

    Takimoto, Noriko; Kusakabe, Takehiro; Horie, Takeo; Miyamoto, Yuki; Tsuda, Motoyuki

    2006-01-01

    We previously identified three genes that encode putative visual cycle proteins that are homologues of retinal G-protein coupled receptor (Ci-opsin3), cellular retinaldehyde-binding protein (Ci-CRALBP) and beta-carotene 15,15'-monooxygenase (Ci-BCO) in the ascidian Ciona intestinalis. Ci-opsin3 and Ci-CRALBP are localized in both ocellus photoreceptor cells and surrounding non-photoreceptor cells in the brain vesicle of the larva. In the present study, we investigated the possible role and evolutionary origin of the BCO/RPE65 family in the visual cycle by analyzing Ci-BCO localization by immunohistochemistry and by identifying a novel gene that encodes a homologue of retinal pigment epithelium-specific 65 kDa protein (Ci-RPE65) in C. intestinalis. In situ hybridization and expressed sequence tag (EST) profiles consistently suggest that Ci-RPE65 is not significantly expressed in the ocellus and brain vesicle of the larva. Ci-RPE65 is expressed in the neural complex, a photoreceptor organ of the adult ascidian, at a level comparable to that of Ci-opsin3 and Ci-CRALBP. Ci-RPE65 is also expressed in various adult tissues, including the gill, body wall and intestine, suggesting that Ci-RPE65 plays a role in addition to that in the visual cycle. In contrast, Ci-BCO is predominantly localized in ocellus photoreceptor cells of the larva. The larval visual cycle seems to use Ci-opsin3 as a photo-isomerase. Our results also suggest that the RPE65-dependent visual cycle is used in the adult photoreceptors of a primitive chordate. PMID:16544957

  18. Spectral Sensitivity of the ctenid spider Cupiennius salei Keys

    PubMed Central

    Zopf, Lydia M.; Schmid, Axel; Fredman, David; Eriksson, Bo Joakim

    2014-01-01

    Summary The spectral sensitivity of adult male Cupiennius salei Keys, a nocturnal hunting spider, was studied in a behavioural test. As known from earlier behavioural tests, C. salei walks towards a black target presented in front of a white background. In this study a black target (size 42 × 70 cm) was presented in a white arena illuminated by monochromatic light in the range of 365 to 695 nm using 19 monochromatic filters (HW in the range of 6 – 10 nm). In the first trial, the transmission of the optical filters was between 40 % and 80%. In a second trial the transmission was reduced to 5%, using a neutral density filter. At the high intensity the spiders showed a spectral sensivity in the range from 380 to 670 nm. In the second trial the animals only showed directed walks if the illumination was in the range of 449 to 599 nm, indicating a lower sensitivity at the margins of the spectral sensitivity. In previous intracellular recordings, the measured spectral sensitivity was between 320 and 620 nm. Interestingly, these results do not completely match the behaviourally tested spectral sensitivity of the photoreceptors, where the sensitivity range is shifted to longer wavelengths. In order to investigate the molecular background of spectral sensitivity, we searched for opsin genes in C. salei. We found three visual opsins that correspond to UV and middle to long wavelength sensitive opsins as described for jumping spiders. PMID:23948480

  19. Non-invasive activation of optogenetic actuators

    NASA Astrophysics Data System (ADS)

    Birkner, Elisabeth; Berglund, Ken; Klein, Marguerita E.; Augustine, George J.; Hochgeschwender, Ute

    2014-03-01

    The manipulation of genetically targeted neurons with light (optogenetics) continues to provide unprecedented avenues into studying the function of the mammalian brain. However, potential translation into the clinical arena faces a number of significant hurdles, foremost among them the need for insertion of optical fibers into the brain to deliver light to opsins expressed on neuronal membranes. In order to overcome these hardware-related problems, we have developed an alternative strategy for delivering light to opsins which does not involve fiber implants. Rather, the light is produced by a protein, luciferase, which oxidizes intravenously applied substrate, thereby emitting bioluminescence. In proof-ofprinciple studies employing a fusion protein of a light-generating luciferase to a light-sensing opsin (luminopsin), we showed that light emitted by Gaussia luciferase is indeed able to activate channelrhodopsin, allowing modulation of neuronal activity when expressed in cultured neurons. Here we assessed applicability of the concept in vivo in mice expressing luminopsins from viral vectors and from genetically engineered transgenes. The experiments demonstrate that intravenously applied substrate reaches neurons in the brain, causing the luciferase to produce bioluminescence which can be imaged in vivo, and that activation of channelrhodopsin by bioluminescence is sufficient to affect behavior. Further developments of such technology based on combining optogenetics with bioluminescence - i.e. combining lightsensing molecules with biologically produced light through luciferases - should bring optogenetics closer to clinical applications.

  20. Non-rod, non-cone photoreception in rodents and teleost fish.

    PubMed

    Foster, Russell G; Hankins, Mark; Lucas, Robert J; Jenkins, Aaron; Muñoz, Marta; Thompson, Stewart; Appleford, Joanne M; Bellingham, James

    2003-01-01

    Until recently, all ocular photoreception was attributed to the rods and cones of the retina. However, studies on mice lacking rod and cone photoreceptors (rd/rd cl), has shown that these mice can still use their eyes to detect light to regulate their circadian rhythms, suppress pineal melatonin, modify locomotor activity and modulate pupil size. In addition, action spectra for some of these responses have characterized a novel opsin/vitamin A-based photopigment with a lambda(max) approximately 480 nm. Electrophysiological studies have shown that a subset of retinal ganglion cells are intrinsically photosensitive, and melanopsin has been proposed as the photopigment mediating these responses to light. In contrast to mammals, an inner retinal photopigment gene has been identified in teleost fish. Vertebrate ancient (VA) opsin forms a photopigment with a lambda(max) between 460-500 nm, and is expressed in a sub-set of retinal horizontal cells, and cells in the amacrine and ganglion cell layers. Electrophysiological analysis suggests that VA opsin horizontal cells are intrinsically photosensitive and encode irradiance information. In contrast to mammals, however, the function of these novel ocular photoreceptors remains unknown. We compare non-rod, non-cone ocular photoreceptors in mammals and fish, and examine the criteria used to place candidate photopigment molecules into a functional context. PMID:14712912

  1. The influence of chromatic background on the photosensitivity of tilapia erythrophores

    PubMed Central

    Chen, Shyh-Chi; Hornsby, Mark A. W.; Robertson, R. Meldrum; Hawryshyn, Craig W.

    2014-01-01

    Summary Non-mammalian vertebrates and invertebrates use extraretinal photoreceptors to detect light and perform diverse non-image-forming functions. Compared to well-studied visual systems, the effect of ambient light conditions on photosensory systems of extraretinal photoreceptors is poorly understood. Chromatophores are photosensitive dermal pigment cells that play an important role in the formation of body color patterns to fit the surrounding environment. Here, we used tilapia erythrophores to investigate the relationship between environmental light and chromatophore photoresponses. All erythrophores from three spectral conditions aggregated their pigment granules in UV/short wavelengths and dispersed in middle/long wavelengths. Unlike retinal visual systems, environmental light did not change the usage of the primary opsins responsible for aggregation and dispersion. In addition, short wavelength-rich and red-shifted background conditions led to an inhibitory effect on erythrophore photoresponses. We suggest that, as extraretinal photoreceptors for non-image-forming functions, chromatophores directly adjust their photoresponse sensitivity via changes in opsin expression levels rather than opsin types when environmental light changes. PMID:24414206

  2. Virally mediated optogenetic excitation and inhibition of pain in freely moving non-transgenic mice

    PubMed Central

    Iyer, Shrivats Mohan; Montgomery, Kate L.; Towne, Chris; Lee, Soo Yeun; Ramakrishnan, Charu; Deisseroth, Karl; Delp, Scott L.

    2014-01-01

    Primary nociceptors are the first neurons involved in the complex processing system that regulates normal and pathological pain1. Our ability to excite and inhibit these neurons has been limited by pharmacological and electrical stimulation constraints; non-invasive excitation and inhibition of these neurons in freely moving non-transgenic animals has not been possible. Here we use an optogenetic2 strategy to bidirectionally control nociceptors of non-transgenic mice. Intra-sciatic nerve injection of adeno-associated viruses encoding an excitatory opsin enabled light-inducible stimulation of acute pain, place aversion, and optogenetically mediated reductions in withdrawal thresholds to mechanical and thermal stimuli. In contrast, viral delivery of an inhibitory opsin enabled light-inducible inhibition of acute pain perception, and reversed mechanical allodynia and thermal hyperalgesia in a model of neuropathic pain. Light was delivered transdermally enabling these behaviors to be induced in freely moving animals. This approach may have utility in basic and translational pain research, and enable rapid drug screening and testing of newly engineered opsins. PMID:24531797

  3. Spectral sensitivity of the ctenid spider Cupiennius salei.

    PubMed

    Zopf, Lydia M; Schmid, Axel; Fredman, David; Eriksson, Bo Joakim

    2013-11-01

    The spectral sensitivity of adult male Cupiennius salei Keys, a nocturnal hunting spider, was studied in a behavioural test. As known from earlier behavioural tests, C. salei will walk towards a black target presented in front of a white background. In this study, a black target (size 42×70 cm) was presented in a white arena illuminated by monochromatic light in the range 365-695 nm using 19 monochromatic filters (half-width in the range 6-10 nm). In the first trial, the transmission of the optical filters was between 40% and 80%. In the second trial, the transmission was reduced to 5% using a neutral density filter. At the high intensity, the spiders showed a spectral sensitivity in the range 380-670 nm. In the second trial, the animals only showed directed walks if the illumination was in the range 449-599 nm, indicating a lower sensitivity at the margins of the spectral sensitivity. In previous intracellular recordings, the measured spectral sensitivity was between 320 and 620 nm. Interestingly, these results do not completely match the behaviourally tested spectral sensitivity of the photoreceptors, where the sensitivity range is shifted to longer wavelengths. In order to investigate the molecular background of spectral sensitivity, we searched for opsin genes in C. salei. We found three visual opsins that correspond to UV and middle to long wavelength sensitive opsins as described for jumping spiders. PMID:23948480

  4. Signatures of Selection and Gene Conversion Associated with Human Color Vision Variation

    PubMed Central

    Verrelli, Brian C.; Tishkoff, Sarah A.

    2004-01-01

    Trichromatic color vision in humans results from the combination of red, green, and blue photopigment opsins. Although color vision genes have been the targets of active molecular and psychophysical research on color vision abnormalities, little is known about patterns of normal genetic variation in these genes among global human populations. The current study presents nucleotide sequence analyses and tests of neutrality for a 5.5-kb region of the X-linked long-wave “red” opsin gene (OPN1LW) in 236 individuals from ethnically diverse human populations. Our analysis of the recombination landscape across OPN1LW reveals an unusual haplotype structure associated with amino acid replacement variation in exon 3 that is consistent with gene conversion. Compared with the absence of OPN1LW amino acid replacement fixation since divergence from chimpanzee, the human population exhibits a significant excess of high-frequency OPN1LW replacements. Our results suggest that subtle changes in L-cone opsin wavelength absorption may have been adaptive during human evolution. PMID:15252758

  5. The origins of colour vision in vertebrates.

    PubMed

    Collin, Shaun P; Trezise, Ann E O

    2004-07-01

    The capacity for colour vision is mediated by the comparison of the signal intensities from photoreceptors of two or more types that differ in spectral sensitivity. Morphological, physiological and molecular analyses of the retina in an agnathan (jawless) fish, the lamprey Geotria australis, may hold important clues to the origins of colour vision in vertebrates. Lampreys are extant representatives of an ancient group of vertebrates, the origins of which are thought to date back to at least the early Cambrian, approximately 540 million years ago. G. australis possesses five photoreceptor types, each with cone-like ultrastructural features and different spectral sensitivities. Recent molecular genetic studies have also revealed that five visual pigment (opsin) genes are expressed in the retina, each of which is orthologous to the major classes of vertebrate opsin genes. These findings reveal that multiple opsin genes originated very early in vertebrate evolution, prior to the separation of the jawed and jawless vertebrate lineages, thereby providing the genetic basis for colour vision in all vertebrates. PMID:15312025

  6. Optogenetics in Mice Performing a Visual Discrimination Task: Measurement and Suppression of Retinal Activation and the Resulting Behavioral Artifact.

    PubMed

    Danskin, Bethanny; Denman, Daniel; Valley, Matthew; Ollerenshaw, Douglas; Williams, Derric; Groblewski, Peter; Reid, Clay; Olsen, Shawn; Waters, Jack

    2015-01-01

    Optogenetic techniques are used widely to perturb and interrogate neural circuits in behaving animals, but illumination can have additional effects, such as the activation of endogenous opsins in the retina. We found that illumination, delivered deep into the brain via an optical fiber, evoked a behavioral artifact in mice performing a visually guided discrimination task. Compared with blue (473 nm) and yellow (589 nm) illumination, red (640 nm) illumination evoked a greater behavioral artifact and more activity in the retina, the latter measured with electrical recordings. In the mouse, the sensitivity of retinal opsins declines steeply with wavelength across the visible spectrum, but propagation of light through brain tissue increases with wavelength. Our results suggest that poor retinal sensitivity to red light was overcome by relatively robust propagation of red light through brain tissue and stronger illumination of the retina by red than by blue or yellow light. Light adaptation of the retina, via an external source of illumination, suppressed retinal activation and the behavioral artifact without otherwise impacting behavioral performance. In summary, long wavelength optogenetic stimuli are particularly prone to evoke behavioral artifacts via activation of retinal opsins in the mouse, but light adaptation of the retina can provide a simple and effective mitigation of the artifact. PMID:26657323

  7. Transcriptome profiling of developing photoreceptor subtypes reveals candidate genes involved in avian photoreceptor diversification.

    PubMed

    Enright, Jennifer M; Lawrence, Karen A; Hadzic, Tarik; Corbo, Joseph C

    2015-03-01

    Avian photoreceptors are a diverse class of neurons, comprised of four single cones, the two members of the double cone, and rods. The signaling events and transcriptional regulators driving the differentiation of these diverse photoreceptors are largely unknown. In addition, many distinctive features of photoreceptor subtypes, including spectral tuning, oil droplet size and pigmentation, synaptic targets, and spatial patterning, have been well characterized, but the molecular mechanisms underlying these attributes have not been explored. To identify genes specifically expressed in distinct chicken (Gallus gallus) photoreceptor subtypes, we developed fluorescent reporters that label photoreceptor subpopulations, isolated these subpopulations by using fluorescence-activated cell sorting, and subjected them to next-generation sequencing. By comparing the expression profiles of photoreceptors labeled with rhodopsin, red opsin, green opsin, and violet opsin reporters, we have identified hundreds of differentially expressed genes that may underlie the distinctive features of these photoreceptor subtypes. These genes are involved in a variety of processes, including phototransduction, transcriptional regulation, cell adhesion, maintenance of intra- and extracellular structure, and metabolism. Of particular note are a variety of differentially expressed transcription factors, which may drive and maintain photoreceptor diversity, and cell adhesion molecules, which may mediate spatial patterning of photoreceptors and act to establish retinal circuitry. These analyses provide a framework for future studies that will dissect the role of these various factors in the differentiation of avian photoreceptor subtypes. PMID:25349106

  8. Transcriptome profiling of developing photoreceptor subtypes reveals candidate genes involved in avian photoreceptor diversification

    PubMed Central

    Enright, Jennifer M.; Lawrence, Karen A.; Hadzic, Tarik; Corbo, Joseph C.

    2015-01-01

    Avian photoreceptors are a diverse class of neurons, comprised of four single cones, the two members of the double cone, and rods. The signaling events and transcriptional regulators driving the differentiation of these diverse photoreceptors are currently unknown. In addition, many distinctive features of photoreceptor subtypes, including spectral tuning, oil droplet size and pigmentation, synaptic targets and spatial patterning, have been well characterized, but the molecular mechanisms underlying these attributes have not been explored. To identify genes specifically expressed in distinct chicken (Gallus gallus) photoreceptor subtypes, we developed fluorescent reporters that label photoreceptor subpopulations, isolated these subpopulations using fluorescence-activated cell sorting and subjected them to next-generation sequencing. By comparing the expression profiles of photoreceptors labeled with rhodopsin, red opsin, green opsin, and violet opsin reporters, we have identified hundreds of differentially expressed genes that may underlie the distinctive features of these photoreceptor subtypes. These genes are involved in a variety of processes, including phototransduction, transcriptional regulation, cell adhesion, maintenance of intra- and extra-cellular structure, and metabolism. Of particular note are a variety of differentially expressed transcription factors, which may drive and maintain photoreceptor diversity, and cell adhesion molecules that may mediate spatial patterning of photoreceptors and act to establish retinal circuitry. These analyses provide a framework for future studies that will dissect the role of these various factors in the differentiation of avian photoreceptor subtypes. PMID:25349106

  9. Making Sense of Optogenetics.

    PubMed

    Guru, Akash; Post, Ryan J; Ho, Yi-Yun; Warden, Melissa R

    2015-10-01

    This review, one of a series of articles, tries to make sense of optogenetics, a recently developed technology that can be used to control the activity of genetically-defined neurons with light. Cells are first genetically engineered to express a light-sensitive opsin, which is typically an ion channel, pump, or G protein-coupled receptor. When engineered cells are then illuminated with light of the correct frequency, opsin-bound retinal undergoes a conformational change that leads to channel opening or pump activation, cell depolarization or hyperpolarization, and neural activation or silencing. Since the advent of optogenetics, many different opsin variants have been discovered or engineered, and it is now possible to stimulate or inhibit neuronal activity or intracellular signaling pathways on fast or slow timescales with a variety of different wavelengths of light. Optogenetics has been successfully employed to enhance our understanding of the neural circuit dysfunction underlying mood disorders, addiction, and Parkinson's disease, and has enabled us to achieve a better understanding of the neural circuits mediating normal behavior. It has revolutionized the field of neuroscience, and has enabled a new generation of experiments that probe the causal roles of specific neural circuit components. PMID:26209858

  10. Ablation of Chop Transiently Enhances Photoreceptor Survival but Does Not Prevent Retinal Degeneration in Transgenic Mice Expressing Human P23H Rhodopsin.

    PubMed

    Chiang, Wei-Chieh; Joseph, Victory; Yasumura, Douglas; Matthes, Michael T; Lewin, Alfred S; Gorbatyuk, Marina S; Ahern, Kelly; LaVail, Matthew M; Lin, Jonathan H

    2016-01-01

    RHO (Rod opsin) encodes a G-protein coupled receptor that is expressed exclusively by rod photoreceptors of the retina and forms the essential photopigment, rhodopsin, when coupled with 11-cis-retinal. Many rod opsin disease -mutations cause rod opsin protein misfolding and trigger endoplasmic reticulum (ER) stress, leading to activation of the Unfolded Protein Response (UPR) signal transduction network. Chop is a transcriptional activator that is induced by ER stress and promotes cell death in response to chronic ER stress. Here, we examined the role of Chop in transgenic mice expressing human P23H rhodopsin (hP23H Rho Tg) that undergo retinal degeneration. With the exception of one time point, we found no significant induction of Chop in these animals and no significant change in retinal degeneration by histology and electrophysiology when hP23H Rho Tg animals were bred into a Chop (-/-) background. Our results indicate that Chop does not play a significant causal role during retinal degeneration in these animals. We suggest that other modules of the ER stress-induced UPR signaling network may be involved photoreceptor disease induced by P23H rhodopsin. PMID:26427410

  11. Optogenetics in Mice Performing a Visual Discrimination Task: Measurement and Suppression of Retinal Activation and the Resulting Behavioral Artifact

    PubMed Central

    Danskin, Bethanny; Denman, Daniel; Valley, Matthew; Ollerenshaw, Douglas; Williams, Derric; Groblewski, Peter; Reid, Clay; Olsen, Shawn; Waters, Jack

    2015-01-01

    Optogenetic techniques are used widely to perturb and interrogate neural circuits in behaving animals, but illumination can have additional effects, such as the activation of endogenous opsins in the retina. We found that illumination, delivered deep into the brain via an optical fiber, evoked a behavioral artifact in mice performing a visually guided discrimination task. Compared with blue (473 nm) and yellow (589 nm) illumination, red (640 nm) illumination evoked a greater behavioral artifact and more activity in the retina, the latter measured with electrical recordings. In the mouse, the sensitivity of retinal opsins declines steeply with wavelength across the visible spectrum, but propagation of light through brain tissue increases with wavelength. Our results suggest that poor retinal sensitivity to red light was overcome by relatively robust propagation of red light through brain tissue and stronger illumination of the retina by red than by blue or yellow light. Light adaptation of the retina, via an external source of illumination, suppressed retinal activation and the behavioral artifact without otherwise impacting behavioral performance. In summary, long wavelength optogenetic stimuli are particularly prone to evoke behavioral artifacts via activation of retinal opsins in the mouse, but light adaptation of the retina can provide a simple and effective mitigation of the artifact. PMID:26657323

  12. Ablation of Chop Transiently Enhances Photoreceptor Survival but Does Not Prevent Retinal Degeneration in Transgenic Mice Expressing Human P23H Rhodopsin

    PubMed Central

    Chiang, Wei-Chieh; Joseph, Victory; Matthes, Michael T.; Lewin, Alfred S.; Gorbatyuk, Marina S.; Ahern, Kelly; LaVail, Matthew M.

    2016-01-01

    RHO (Rod opsin) encodes a G-protein coupled receptor that is expressed exclusively by rod photoreceptors of the retina and forms the essential photopigment, rhodopsin, when coupled with 11-cis-retinal. Many rod opsin disease mutations cause rod opsin protein misfolding and trigger endoplasmic reticulum (ER) stress, leading to activation of the Unfolded Protein Response (UPR) signal transduction network. Chop is a transcriptional activator that is induced by ER stress and promotes cell death in response to chronic ER stress. Here, we examined the role of Chop in transgenic mice expressing human P23H rhodopsin (hP23H Rho Tg) that undergo retinal degeneration. With the exception of one time point, we found no significant induction of Chop in these animals and no significant change in retinal degeneration by histology and electrophysiology when hP23H Rho Tg animals were bred into a Chop?/? background. Our results indicate that Chop does not play a significant causal role during retinal degeneration in these animals. We suggest that other modules of the ER stress-induced UPR signaling network may be involved photoreceptor disease induced by P23H rhodopsin. PMID:26427410

  13. Differential light-induced responses in sectorial inherited retinal degeneration.

    PubMed

    Ramon, Eva; Cordomí, Arnau; Aguilà, Mònica; Srinivasan, Sundaramoorthy; Dong, Xiaoyun; Moore, Anthony T; Webster, Andrew R; Cheetham, Michael E; Garriga, Pere

    2014-12-26

    Retinitis pigmentosa (RP) is a group of genetically and clinically heterogeneous inherited degenerative retinopathies caused by abnormalities of photoreceptors or retinal pigment epithelium in the retina leading to progressive sight loss. Rhodopsin is the prototypical G-protein-coupled receptor located in the vertebrate retina and is responsible for dim light vision. Here, novel M39R and N55K variants were identified as causing an intriguing sector phenotype of RP in affected patients, with selective degeneration in the inferior retina. To gain insights into the molecular aspects associated with this sector RP phenotype, whose molecular mechanism remains elusive, the mutations were constructed by site-directed mutagenesis, expressed in heterologous systems, and studied by biochemical, spectroscopic, and functional assays. M39R and N55K opsins had variable degrees of chromophore regeneration when compared with WT opsin but showed no gross structural misfolding or altered trafficking. M39R showed a faster rate for transducin activation than WT rhodopsin with a faster metarhodopsinII decay, whereas N55K presented a reduced activation rate and an altered photobleaching pattern. N55K also showed an altered retinal release from the opsin binding pocket upon light exposure, affecting its optimal functional response. Our data suggest that these sector RP mutations cause different protein phenotypes that may be related to their different clinical progression. Overall, these findings illuminate the molecular mechanisms of sector RP associated with rhodopsin mutations. PMID:25359768

  14. Volvoxrhodopsin, a light-regulated sensory photoreceptor of the spheroidal green alga Volvox carteri.

    PubMed Central

    Ebnet, E; Fischer, M; Deininger, W; Hegemann, P

    1999-01-01

    Somatic cells of the multicellular alga Volvox carteri contain a visual rhodopsin that controls the organism's phototactic behavior via two independent photoreceptor currents. Here, we report the identification of an opsinlike gene, designated as volvoxopsin (vop). The encoded protein exhibits homologies to the opsin of the unicellular alga Chlamydomonas reinhardtii (chlamyopsin) and to the entire animal opsin family, thus providing new perspectives on opsin evolution. Volvoxopsin accumulates within the eyes of somatic cells. However, the vop transcript is detectable only in the reproductive eyeless gonidia and embryos. vop mRNA levels increase 400-fold during embryogenesis, when embryos develop in darkness, whereas the vop transcript does not accumulate when embryos develop in the light. An antisense transformant, T3, was generated. This transformant produces 10 times less volvoxopsin than does the wild type. In T3, the vop transcript is virtually absent, whereas the antisense transcript is predominant and light regulated. It follows that vop expression is under light-dependent transcriptional control but that volvoxopsin itself is not the regulatory photoreceptor. Transformant T3 is phototactic, but its phototactic sensitivity is reduced 10-fold relative to the parental wild-type strain HK10. Thus, we offer definitive genetic evidence that a rhodopsin serves as the photoreceptor for phototaxis in a green alga. PMID:10449581

  15. Differential Light-induced Responses in Sectorial Inherited Retinal Degeneration*

    PubMed Central

    Ramon, Eva; Cordomí, Arnau; Aguilà, Mònica; Srinivasan, Sundaramoorthy; Dong, Xiaoyun; Moore, Anthony T.; Webster, Andrew R.; Cheetham, Michael E.; Garriga, Pere

    2014-01-01

    Retinitis pigmentosa (RP) is a group of genetically and clinically heterogeneous inherited degenerative retinopathies caused by abnormalities of photoreceptors or retinal pigment epithelium in the retina leading to progressive sight loss. Rhodopsin is the prototypical G-protein-coupled receptor located in the vertebrate retina and is responsible for dim light vision. Here, novel M39R and N55K variants were identified as causing an intriguing sector phenotype of RP in affected patients, with selective degeneration in the inferior retina. To gain insights into the molecular aspects associated with this sector RP phenotype, whose molecular mechanism remains elusive, the mutations were constructed by site-directed mutagenesis, expressed in heterologous systems, and studied by biochemical, spectroscopic, and functional assays. M39R and N55K opsins had variable degrees of chromophore regeneration when compared with WT opsin but showed no gross structural misfolding or altered trafficking. M39R showed a faster rate for transducin activation than WT rhodopsin with a faster metarhodopsinII decay, whereas N55K presented a reduced activation rate and an altered photobleaching pattern. N55K also showed an altered retinal release from the opsin binding pocket upon light exposure, affecting its optimal functional response. Our data suggest that these sector RP mutations cause different protein phenotypes that may be related to their different clinical progression. Overall, these findings illuminate the molecular mechanisms of sector RP associated with rhodopsin mutations. PMID:25359768

  16. Melanopsin mediates light-dependent relaxation in blood vessels

    PubMed Central

    Sikka, Gautam; Hussmann, G. Patrick; Pandey, Deepesh; Cao, Suyi; Hori, Daijiro; Park, Jong Taek; Steppan, Jochen; Kim, Jae Hyung; Barodka, Viachaslau; Myers, Allen C.; Santhanam, Lakshmi; Nyhan, Daniel; Halushka, Marc K.; Koehler, Raymond C.; Snyder, Solomon H.; Shimoda, Larissa A.; Berkowitz, Dan E.

    2014-01-01

    Melanopsin (opsin4; Opn4), a non-image-forming opsin, has been linked to a number of behavioral responses to light, including circadian photo-entrainment, light suppression of activity in nocturnal animals, and alertness in diurnal animals. We report a physiological role for Opn4 in regulating blood vessel function, particularly in the context of photorelaxation. Using PCR, we demonstrate that Opn4 (a classic G protein-coupled receptor) is expressed in blood vessels. Force-tension myography demonstrates that vessels from Opn4−/− mice fail to display photorelaxation, which is also inhibited by an Opn4-specific small-molecule inhibitor. The vasorelaxation is wavelength-specific, with a maximal response at ∼430–460 nm. Photorelaxation does not involve endothelial-, nitric oxide-, carbon monoxide-, or cytochrome p450-derived vasoactive prostanoid signaling but is associated with vascular hyperpolarization, as shown by intracellular membrane potential measurements. Signaling is both soluble guanylyl cyclase- and phosphodiesterase 6-dependent but protein kinase G-independent. β-Adrenergic receptor kinase 1 (βARK 1 or GRK2) mediates desensitization of photorelaxation, which is greatly reduced by GRK2 inhibitors. Blue light (455 nM) regulates tail artery vasoreactivity ex vivo and tail blood blood flow in vivo, supporting a potential physiological role for this signaling system. This endogenous opsin-mediated, light-activated molecular switch for vasorelaxation might be harnessed for therapy in diseases in which altered vasoreactivity is a significant pathophysiologic contributor. PMID:25404319

  17. Identification of DES1 as a Vitamin A Isomerase in Müller Glial Cells of the Retina

    PubMed Central

    Kaylor, Joanna J.; Yuan, Quan; Cook, Jeremy; Sarfare, Shanta; Makshanoff, Jacob; Miu, Anh; Kim, Anita; Kim, Paul; Habib, Samer; Roybal, C. Nathaniel; Xu, Tongzhou; Nusinowitz, Steven; Travis, Gabriel H.

    2012-01-01

    Absorption of a light particle by an opsin-pigment causes photoisomerization of its retinaldehyde chromophore. Restoration of light sensitivity to the resulting apo-opsin requires chemical re-isomerization of the photobleached chromophore. This is carried out by a multistep enzyme pathway called the visual cycle. Accumulating evidence suggests the existence of an alternate visual cycle for regenerating opsins in daylight. Here, we identified dihydroceramide desaturase-1 (DES1) as a retinol isomerase and an excellent candidate for isomerase-2 in this alternate pathway. DES1 is expressed in retinal Müller cells where it co-immunoprecipitates with cellular retinaldehyde binding protein (CRALBP). Adenoviral gene therapy with DES1 partially rescued the biochemical and physiological phenotypes in rpe65 −/− mice lacking isomerohydrolase (isomerase-1). Knockdown of DES1 expression by RNA-interference concordantly reduced isomerase-2 activity in cultured Müller cells. Purified DES1 possessed very high isomerase-2 activity in the presence of appropriate cofactors, suggesting that DES1 by itself is sufficient for isomerase activity. PMID:23143414

  18. Beyond spectral tuning: human cone visual pigments adopt different transient conformations for chromophore regeneration.

    PubMed

    Srinivasan, Sundaramoorthy; Cordom, Arnau; Ramon, Eva; Garriga, Pere

    2016-03-01

    Human red and green visual pigments are seven transmembrane receptors of cone photoreceptor cells of the retina that mediate color vision. These pigments share a very high degree of homology and have been assumed to feature analogous structural and functional properties. We report on a different regeneration mechanism among red and green cone opsins with retinal analogs using UV-Vis/fluorescence spectroscopic analyses, molecular modeling and site-directed mutagenesis. We find that photoactivated green cone opsin adopts a transient conformation which regenerates via an unprotonated Schiff base linkage with its natural chromophore, whereas red cone opsin forms a typical protonated Schiff base. The chromophore regeneration kinetics is consistent with a secondary retinal uptake by the cone pigments. Overall, our findings reveal, for the first time, structural differences in the photoactivated conformation between red and green cone pigments that may be linked to their molecular evolution, and support the proposal of secondary retinal binding to visual pigments, in addition to binding to the canonical primary site, which may serve as a regulatory mechanism of dark adaptation in the phototransduction process. PMID:26387074

  19. Retinal Pathology of a Patient with Goldmann-Favre Syndrome

    PubMed Central

    Bonilha, Vera L.; Fishman, Gerald A.; Rayborn, Mary E.; Hollyfield, Joe G.

    2013-01-01

    Purpose To define the retinal pathology in an 88 year-old male affected with Goldmann-Favre syndrome with a 2bp 5 A>C splice site mutation in the NR2E3 gene. Methods Retinal tissue from the macula and periphery was processed for immunohistochemistry. Perimacular retina was processed for transmission electron microscopy. Cryosections were studied by indirect immunofluorescence, using well-characterized antibodies to rhodopsin, cone cytoplasm, and cone opsins. The affected donor eye was compared to a postmortem matched normal eye. Results The retina was highly disorganized without laminar organization. The RPE was discontinuous in some perimacular regions. Large (>1mm) spherical electrondense melanosomes were observed in the RPE and choroid by TEM. Rods were virtually absent in the affected retina. Cones were present in the macula, but were mostly absent from the retinal periphery. In addition, cone rosettes were observed in the perimacular area. Both red/green and blue cone opsins were distributed along the entire cellular expanse of the cone photoreceptors in the affected eye, but were restricted to the cone outer segments in the control retina. Conclusions The histological data obtained from the retina of an elderly male patient with Goldmann-Favre syndrome showed an absence of rods and abnormal distribution of red/green and blue cone opsins. PMID:19852574

  20. Identification and characterization of visual pigments in caecilians (Amphibia: Gymnophiona), an order of limbless vertebrates with rudimentary eyes.

    PubMed

    Mohun, S M; Davies, W L; Bowmaker, J K; Pisani, D; Himstedt, W; Gower, D J; Hunt, D M; Wilkinson, M

    2010-10-15

    In comparison with the other amphibian orders, the Anura (frogs) and Urodela (salamanders), knowledge of the visual system of the snake-like Gymnophiona (caecilians) is relatively sparse. Most caecilians are fossorial with, as far as is known any surface activity occurring mainly at night. They have relatively small, poorly developed eyes and might be expected to possess detectable changes in the spectral sensitivity of their visual pigments. Microspectrophotometry was used to determine the spectral sensitivities of the photoreceptors in three species of caecilian, Rhinatrema bivittatum, Geotrypetes seraphini and Typhlonectes natans. Only rod opsin visual pigment, which may be associated with scotopic (dim light) vision when accompanied by other 'rod-specific' components of the phototransduction cascade, was found to be present. Opsin sequences were obtained from the eyes of two species of caecilian, Ichthyophis cf. kohtaoensis and T. natans. These rod opsins were regenerated in vitro with 11-cis retinal to give pigments with spectral sensitivity peaks close to 500 nm. No evidence for cone photoreception, associated with diurnal and colour vision, was detected using molecular and physiological methods. Additionally, visual pigments are short-wavelength shifted in terms of the maximum absorption of light when compared with other amphibian lineages. PMID:20889838

  1. Cone-Specific Promoters for Gene Therapy of Achromatopsia and Other Retinal Diseases.

    PubMed

    Ye, Guo-Jie; Budzynski, Ewa; Sonnentag, Peter; Nork, T Michael; Sheibani, Nader; Gurel, Zafer; Boye, Sanford L; Peterson, James J; Boye, Shannon E; Hauswirth, William W; Chulay, Jeffrey D

    2016-01-01

    Adeno-associated viral (AAV) vectors containing cone-specific promoters have rescued cone photoreceptor function in mouse and dog models of achromatopsia, but cone-specific promoters have not been optimized for use in primates. Using AAV vectors administered by subretinal injection, we evaluated a series of promoters based on the human L-opsin promoter, or a chimeric human cone transducin promoter, for their ability to drive gene expression of green fluorescent protein (GFP) in mice and nonhuman primates. Each of these promoters directed high-level GFP expression in mouse photoreceptors. In primates, subretinal injection of an AAV-GFP vector containing a 1.7-kb L-opsin promoter (PR1.7) achieved strong and specific GFP expression in all cone photoreceptors and was more efficient than a vector containing the 2.1-kb L-opsin promoter that was used in AAV vectors that rescued cone function in mouse and dog models of achromatopsia. A chimeric cone transducin promoter that directed strong GFP expression in mouse and dog cone photoreceptors was unable to drive GFP expression in primate cones. An AAV vector expressing a human CNGB3 gene driven by the PR1.7 promoter rescued cone function in the mouse model of achromatopsia. These results have informed the design of an AAV vector for treatment of patients with achromatopsia. PMID:26603570

  2. Beta-ionone activates and bleaches visual pigment in salamander photoreceptors.

    PubMed

    Isayama, Tomoki; McCabe England, S L; Crouch, R K; Zimmerman, A L; Makino, C L

    2009-01-01

    Vision begins with photoisomerization of 11-cis retinal to the all-trans conformation within the chromophore-binding pocket of opsin, leading to activation of a biochemical cascade. Release of all-trans retinal from the binding pocket curtails but does not fully quench the ability of opsin to activate transducin. All-trans retinal and some other analogs, such as beta-ionone, enhance opsin's activity, presumably on binding the empty chromophore-binding pocket. By recording from isolated salamander photoreceptors and from patches of rod outer segment membrane, we now show that high concentrations of beta-ionone suppressed circulating current in dark-adapted green-sensitive rods by inhibiting the cyclic nucleotide-gated channels. There were also decreases in circulating current and flash sensitivity, and accelerated flash response kinetics in dark-adapted blue-sensitive (BS) rods and cones, and in ultraviolet-sensitive cones, at concentrations too low to inhibit the channels. These effects persisted in BS rods even after incubation with 9-cis retinal to ensure complete regeneration of their visual pigment. After long exposures to high concentrations of beta-ionone, recovery was incomplete unless 9-cis retinal was given, indicating that visual pigment had been bleached. Therefore, we propose that beta-ionone activates and bleaches some types of visual pigments, mimicking the effects of light. PMID:19500430

  3. Tracking the primary photoconversion events in rhodopsins by ultrafast optical spectroscopy.

    PubMed

    Polli, D; Rivalta, I; Nenov, A; Weingart, O; Garavelli, M; Cerullo, G

    2015-02-01

    Opsins are a broad class of photoactive proteins, found in all classes of living beings from bacteria to higher animals, which work either as light-driven ion pumps or as visual pigments. The photoactive function in opsins is triggered by the ultrafast isomerization of the retinal chromophore around a specific carbon double bond, leading to a highly distorted, spectrally red-shifted photoproduct. Understanding, by either experimental or computational methods, the time course of this photoisomerization process is of utmost importance, both for its biological significance and because opsin proteins are the blueprint for molecular photoswitches. This paper focuses on the ultrafast 11-cis to all-trans isomerization in visual rhodopsins, and has a twofold goal: (i) to review the most recent experimental and computational efforts aimed at exposing the very early phases of photoconversion; and (ii) discuss future advanced experiments and calculations that will allow an even deeper understanding of the process. We present high time resolution pump-probe data, enabling us to follow the wavepacket motion through the conical intersection connecting excited and ground states, as well as femtosecond stimulated Raman scattering data allowing us to track the subsequent structural evolution until the first stable all-trans photoproduct is reached. We conclude by introducing computational results for two-dimensional electronic spectroscopy, which has the potential to provide even greater detail on the evolution of the electronic structure of retinal during the photoisomerization process. PMID:25429920

  4. Rhodopsin Photoactivation Dynamics Revealed by Quasi-Elastic Neutron Scattering

    NASA Astrophysics Data System (ADS)

    Bhowmik, Debsindhu; Shrestha, Utsab; Perera, Suchhithranga M. C. D.; Chawla, Udeep; Mamontov, Eugene; Brown, Michael; Chu, Xiang-Qiang

    2015-03-01

    Rhodopsin is a G-protein-coupled receptor (GPCR) responsible for vision. During photoactivation, the chromophore retinal dissociates from protein yielding the opsin apoprotein. What are the changes in protein dynamics that occur during the photoactivation process? Here, we studied the microscopic dynamics of dark-state rhodopsin and the ligand-free opsin using quasielastic neutron scattering (QENS). The QENS technique tracks individual hydrogen atom motion because of the much higher neutron scattering cross-section of hydrogen than other atoms. We used protein with CHAPS detergent hydrated with heavy water. The activation of proteins is confirmed at low temperatures up to 300 K by mean-square displacement (MSD) analysis. The QENS experiments at temperatures ranging from 220 K to 300 K clearly indicate an increase in protein dynamic behavior with temperature. The relaxation time for the ligand-bound protein rhodopsin is faster compared to opsin, which can be correlated with the photoactivation. Moreover, the protein dynamics are orders of magnitude slower than the accompanying CHAPS detergent, which unlike protein, manifests localized motions.

  5. Evolution of Vision

    NASA Astrophysics Data System (ADS)

    Ostrovsky, Mikhail

    The evolution of photoreception, giving rise to eye, offers a kaleidoscopic view on selection acting at both the organ and molecular levels. The molecular level is mainly considered in the lecture. The greatest progress to date has been made in relation to the opsin visual pigments. Opsins appeared before eyes did. Two- and three-dimensional organization for rhodopsin in the rod outer segment disk membrane, as well as molecular mechanisms of visual pigments spectral tuning, photoisomerization and also opsin as a G-protein coupled receptor are considered. Molecular mechanisms of visual pigments spectral tuning, namely switching of chromophore (physiological time scale) and amino acid changes in the chromophore site of opsin (evolutionary time scale) is considered in the lecture. Photoisomerization of rhodopsin chromophore, 11-cis retinal is the only photochemical reaction in vision. The reaction is extemely fast (less that 200 fs) and high efficient (. is 0.65). The rhodopsin photolysis and kinetics of the earlier products appearance, photo- and bathorhodopsin, is considered. It is known that light is not only a carrier of information, but also a risk factor of damage to the eye. This photobiological paradox of vision is mainly due to the nature of rhodopsin chromophore. Photooxidation is the base of the paradox. All factors present in the phototrceptor cells to initiate free-radical photooxidation: photosensitizers, oxygen and substrates of oxidation: lipids and proteins (opsin). That is why photoprotective system of the eye structures appeared in the course of evolution. Three lines of protective system to prevent light damage to the retina and retina pigment epithelium is known: permanent renewal of rod and cone outer segment, powerful antioxidant system and optical media as cut-off filters where the lens is a key component. The molecular mechanisms of light damage to the eye and photoprotective system of the eye is considered in the lecture. The molecular mechanisms of phototransduction in vertebrates eye is also briefly considered in the lecture. Evolution of vision is an enormous subject for thought and investigation. In the postgenomic era evolutionary molecular physiology as a whole and evolutionary molecular physiology of vision can be considered as a key approach for understanding how genome is working.

  6. Restoration of AMPTE/CHEM Data

    NASA Astrophysics Data System (ADS)

    Vandegriff, J.

    2006-12-01

    The highest time resolution digital data for the Charge-Energy-Mass (CHEM) Spectrometer instrument on the Active Magnetospheric Particle Explorers (AMPTE) mission has been restored and made available online at http://sd-www.jhuapl.edu/AMPTE. During the period 1984 to the beginning of 1989, CHEM measured the mass and charge-state compositions as well as the energy spectra and pitch-angle distributions of all major ions from H through Fe with energies from 0.3 to 300 keV/charge. The data cadence is less than one minute. With the spacecraft in a low inclination orbit passing through the magnetopause and well into the magnetosphere, many samplings of the ring-current were made. The data itself consist of so-called PHA (pulse height analysis) events, in which the energy per charge, time of flight, and total energy are measured for each particle. The original Fortran software for calculating fluxes from the PHA events has been ported from VMS to modern Unix Fortran compilers. The CHEM data will be very useful in conjunction with the upcoming Radiation Belt Storm Probe (RBSP) mission, which is part of NASA's Living With a Star (LWS) initiative, and this restoration effort was funded through the LWS Targeted Research and Technology (TR&T) program.

  7. Global-scale Observations of the Limb and Disk (GOLD): Science Objectives

    NASA Astrophysics Data System (ADS)

    Eastes, R.; McClintock, W.; Aksnes, A.; Anderson, D.; Andersson, L.; Burns, A.; Codrescu, M.; Daniell, R.; Eparvier, F.; Harvey, J.; Krywonos, A.; Lumpe, J.; Prölss, G.; Richmond, A.; Rusch, D.; Solomon, S.; Strickland, D.; Woods, T.

    2006-12-01

    Developing the scientific understanding necessary to address aspects of the connected Sun-Earth system that directly affect life and society is a challenge for the scientific community in general and the Living With a Star (LWS) program in particular. Far Ultraviolet (FUV) observations from satellites have made vital contributions to this understanding. The Global-scale Observations of the Earth and Disk (GOLD) experiment, a FUV imager being considered for flight on a geostationary (GEO) satellite as part of the upcoming Radiation Belt Storm Probes (RBSP) mission, will, due to its capabilities and the orbit, provide data needed to answer outstanding science questions. GOLD will provide the first global-scale measurements of temperatures in the lower thermosphere. In addition it will provide more familiar measurements, such as the atomic oxygen to molecular nitrogen (O/N2) ratio. From GEO, GOLD will almost continuously observe the same hemisphere and provide data at all local times each day. These data will allow us to: (1) understand the global-scale response of the thermosphere and ionosphere to geomagnetic and solar forcing; (2) understand the global-scale tidal amplitude and phase variations; and (3) better understand the causes of small-scale ionospheric density irregularities. Such observations are necessary for achieving the LWS program objectives.

  8. O-bearing Molecules in Carbon-rich Proto-Planetary Objects: Study of CRL 618

    NASA Astrophysics Data System (ADS)

    Herpin, F.; Cernicharo, J.

    2001-08-01

    We present ISO LWS observations of the proto-planetary nebula CRL 618, AFGL 618 a star evolving very fast to the planetary nebula stage. In addition to the lines of COd, COt, HCN and HNC, we report on the detection of water and OH emission together with the fine structure lines of [OI] at 63 and 145 microns. We suggest that O-bearing species other than CO are produced in the innermost region of the circumstellar envelope. The UV photons from the central star photodissociate most of the molecular species produced in the AGB phase and allow a chemistry dominated by standard ion-neutral reactions. Not only allow these reactions the formation of O-bearing species, but they also modify the abundances of C-rich molecules like HCN and HNC for which we found an abundance ratio of ? 1, much lower than in AGB stars. The molecular abundances in the different regions of the circumstellar envelope have been derived from radiative transfer models and our knowledge of its physical structure. Furthermore, we compare ISO LWS observations of three C-rich objects typical of each step of the fast transition of an AGB star to the Planetay Nebula stage: CRL 2688, AFGL 2688 a very young Proto-Planetary Nebula, CRL 618 a Proto-Planetary Nebula, and NGC 7027 a young Planetary Nebula.

  9. [Current views on vision of mammals].

    PubMed

    Khokhlova, T V

    2012-01-01

    In the review, research data are presented on mammals' vision including visual pigments, color and contrast vision, and visual behaviour in different species. It is shown that in course of evolution mammals were gradually losing the elements of daylight cone vision system that are typical of other vertebrates. In monotremes, visual pigments SWS2 (cone blue-sensitive 2) and MWS/LWS (green/red-sensitive) are still present, as well as rod RH1. Theria, except some primates, also have two cone visual pigments: SWS1 (ultraviolet/violet or blue-sensitive 1) and MWS/LWS along with rod RH1. Humans and some other higher primates evolved the new visual pigment, MWS, and acquired trichromatic vision. Marine mammals (cetaceans and pinnipeds) and some species of other orders have lost also the visual pigment SWS1, probably due to specificity of processing the information received by these cones. Current view on mammals' vision with two cone pigments and rods is presented. Data on maximum spectral sensitivity of visual pigments in different species and orders are given along with data on spatial contrast sensation. High visual acuity has been acquired by ungulates, artiodactyls, and primates, while the highest one--by humans with their specialized fovea. PMID:23330397

  10. Dust processing in the Carina nebula region

    NASA Astrophysics Data System (ADS)

    Onaka, Takashi; Mori, Tamami I.; Okada, Yoko

    2015-10-01

    Dust processing in the Carina nebula is investigated based on mid- to far-infrared spectroscopy with Infrared Space Observatory (ISO). Mapping observations over a central 40‧ ×20‧ area of the nebula with PHT-S, SWS, and LWS onboard ISO not only reveal spectroscopically that the mid-infrared unidentified infrared (UIR) bands at 6.2, 7.7, 8.6, and 11.3 μm are absent in the ionized region, but also indicate that the 11.3 μm may behave differently from the other three UIR bands near the edge of the ionized region, suggesting a variation either in the size distribution or in the ionization fraction of the band carriers. The correlation of [NII]122 μm and [SiII]35 μm line emissions observed with SWS and LWS is reinvestigated based on the recent atomic data as well as the latest cosmic abundance, suggesting that a large fraction (> 70%), if not all, of silicon returns to the gas phase in the Carina nebula, suggesting that silicates cannot survive under harsh conditions, such as massive star-forming regions. The present observations clearly show dust processing taking place in active regions in the Galaxy.

  11. The Objectives of NASA's Living with a Star Space Environment Testbed

    NASA Technical Reports Server (NTRS)

    Barth, Janet L.; LaBel, Kenneth A.; Brewer, Dana; Kauffman, Billy; Howard, Regan; Griffin, Geoff; Day, John H. (Technical Monitor)

    2001-01-01

    NASA is planning to fly a series of Space Environment Testbeds (SET) as part of the Living With A Star (LWS) Program. The goal of the testbeds is to improve and develop capabilities to mitigate and/or accommodate the affects of solar variability in spacecraft and avionics design and operation. This will be accomplished by performing technology validation in space to enable routine operations, characterize technology performance in space, and improve and develop models, guidelines and databases. The anticipated result of the LWS/SET program is improved spacecraft performance, design, and operation for survival of the radiation, spacecraft charging, meteoroid, orbital debris and thermosphere/ionosphere environments. The program calls for a series of NASA Research Announcements (NRAs) to be issued to solicit flight validation experiments, improvement in environment effects models and guidelines, and collateral environment measurements. The selected flight experiments may fly on the SET experiment carriers and flights of opportunity on other commercial and technology missions. This paper presents the status of the project so far, including a description of the types of experiments that are intended to fly on SET-1 and a description of the SET-1 carrier parameters.

  12. Evolution of the stimulated Raman scattering instability in two-dimensional particle-in-cell simulations

    SciTech Connect

    Masson-Laborde, P. E.; Casanova, M.; Loiseau, P.; Rozmus, W.; Peng, Z.; Pesme, D.; Hueller, S.; Chapman, T.; Bychenkov, V. Yu.

    2010-09-15

    In the following work, we analyze one-dimensional (1D) and two-dimensional (2D) full particle-in-cell simulations of stimulated Raman scattering (SRS) and study the evolution of Langmuir waves (LWs) in the kinetic regime. It is found that SRS reflectivity becomes random due to a nonlinear frequency shift and that the transverse modulations of LWs are induced by (i) the Weibel instability due to the current of trapped particles and (ii) the trapped particle modulational instability (TPMI) [H. Rose, Phys. Plasmas 12, 12318 (2005)]. Comparisons between 1D and 2D cases indicate that the nonlinear frequency shift is responsible for the first saturation of SRS. After this transient interval of first saturation, 2D effects become important: a strong side-scattering of the light, caused by these transverse modulations of the LW and the presence of a nonlinear frequency shift, is observed together with a strong transverse diffusion. This leads to an increase of the Landau damping rate of the LW, contributing to the limiting of Raman backscattering. A model is developed that reproduces the transverse evolution of the magnetic field due to trapped particles. Based on a simple 1D hydrodynamic model, the growth rate for the Weibel instability of the transverse electrostatic mode and magnetic field is estimated and found to be close to the TPMI growth rate [H. Rose et al., Phys. Plasmas 15, 042311 (2008)].

  13. Draft Science Topics for ROSES 2017 NASA Living with a Star Targeted Research and Technology Program

    NASA Astrophysics Data System (ADS)

    Linton, Mark; Zesta, Eftyhia

    2016-05-01

    The NASA Living with a Star Targeted Research and Technology (LWS TR&T) steering committee would like to present a draft of the TR&T science topics being developed for ROSES 2017 to the science community for comment at this conference. These topics will be drafted before this conference at the May 2016 steering committee meeting, based on community input and LWS TR&T goals. The committee is seeking community comment on these draft topics before the topics are finalized at the committee's summer meeting and sent to NASA in the committee's 2016 report. The full text of these draft topics will be presented at this poster, and we aim to hold a town hall for community discussion of these topics during this conference. Please see http://lwstrt.gsfc.nasa.gov for more information on the TR&T program, the steering committee and the draft topics.This work was supported by the NASA Living with a Star program.

  14. The linewidth-size scaling law of molecular gas in the Galaxy

    NASA Astrophysics Data System (ADS)

    Falgarone, Edith G.; McKee, Christopher

    2015-08-01

    The origin of the linewidth-size (LWS) scaling law, first noticed by Larson three decades ago and ascribed to turbulence, is still a highly debated issue. Not unexpectedly, its properties depend on the environment and on the line tracer used.When the optically thick 12CO (J=1-0) line is used, a specific medium is sampled: the translucent molecular gas of moderate density that builds up the bulk of the molecular interstellar medium in galaxies like ours. The sensitivity of the 12CO line to this gas is such that the LWS is found to hold over almost five orders of magnitude in lengthscale, although with a considerable scatter (1 dex). It reveals an invariant of the cascade, the specific kinetic energy tranfer rate. It also appears to split into two regimes, depending on the gas mass surface density: below a given threshold that is proposed to be linked to the galactic dynamics, it bears the signature of a turbulent cascade, while above that threshold, the scaling law is ascribed to virial balance between turbulent energy and gravity. Large deviations from the scaling law are observed at small scales where signatures of turbulent intermittency may be present.

  15. Lamb Wave-Based Acoustic Radiation Force-Driven Particle Ring Formation Inside a Sessile Droplet.

    PubMed

    Destgeer, Ghulam; Ha, Byunghang; Park, Jinsoo; Sung, Hyung Jin

    2016-04-01

    We demonstrate an acoustofluidic device using Lamb waves (LWs) to manipulate polystyrene (PS) microparticles suspended in a sessile droplet of water. The LW-based acoustofluidic platform used in this study is advantageous in that the device is actuated over a range of frequencies without changing the device structure or electrode pattern. In addition, the device is simple to operate and cheap to fabricate. The LWs, produced on a piezoelectric substrate, attenuate inside the fluid and create acoustic streaming flow (ASF) in the form of a poloidal flow with toroidal vortices. The PS particles experience direct acoustic radiation force (ARF) in addition to being influenced by the ASF, which drive the concentration of particles to form a ring. This phenomenon was previously attributed to the ASF alone, but the present experimental results confirm that the ARF plays an important role in forming the particle ring, which would not be possible in the presence of only the ASF. We used a range of actuation frequencies (45-280 MHz), PS particle diameters (1-10 μm), and droplet volumes (5, 7.5, and 10 μL) to experimentally demonstrate this phenomenon. PMID:26937678

  16. Fermentation of liquid coproducts and liquid compound diets: Part 2. Effects on pH, acid-binding capacity, organic acids and ethanol during a 6-day storage period.

    PubMed

    Scholten, R H; Rijnen, M M; Schrama, J W; Boer, H; van der Peet-Schwering, C M; Den Hartog, L A; Vesseur, P C; Verstegen, M W

    2001-06-01

    The effects of a 6-day storage period on changes in pH, acid-binding capacity, level of organic acids and ethanol of three liquid coproducts [liquid wheat starch (LWS), mashed potato steam peel (PSP) and cheese whey (CW)] and two liquid compound diets [liquid grower diet (LGD) and liquid finisher diet (LFD)] were studied. All products, except LWS, showed a significant decrease in pH and acid-binding capacity during storage. At the end of the storage period, all products reached a pH of between 3.5 and 3.9. In general, it can be concluded that the lactic acid content, and to a lesser extent the acetic acid content, increased dramatically during storage. In contrast, the ethanol content increased significantly in the liquid compound diets only. The pattern of changes in pH and organic acids during the 6-day storage period was different between the liquid coproducts and the liquid compound diets. At the start of storage, liquid coproducts are already in the 'middle' of the fermentation process, while liquid compound diets need approximately 24-36 h before fermentation begins. Consequently, in practice a different approach to obtain fermented diets is needed for liquid coproducts and liquid compound diets. PMID:11686780

  17. Current Status on NASA's Living With a Star Program

    NASA Astrophysics Data System (ADS)

    Guhathakurta, M.; Stcyr, O. C.

    Living With a Star is a space weather-focused and application-driven research program. Its goal is to develop the scientific understanding necessary to effectively address those aspects of the connected Sun-Earth system that directly affect life and society. This program is part of the Sun-Earth Connection (SEC) theme within the office of Space Science. Living With a Star is a cross-cutting program whose goals and objectives have the following links to each of the four NASA Strategic Enterprises: Space Science: LWS quantifies the physics, dynamics, and behavior of the only stellar/planetary system we can see at a close distance. Earth Science: LWS improves understanding of the effects of solar variability and disturbances on terrestrial climate change. Human Space Flight: LWS provides data and scientific understanding required for advanced warning of energetic particle events that affect the safety of humans in space. Aeronautics and Space Transportation: LWS provides detailed characterization of radiation environments useful in the design of more reliable electronic components for air and space transportation system. LWS Program is implemented by a sequence of inter-related science missions, space environment test bed and targeted research and technology. 1) A space weather research network of spacecraft will provide continuous observations of the Sun-Earth system for interlocking, dual use, scientific and applications research. Flight of a Solar Dynamics Observatory (launch 2008) to: - Probe solar interior, especially region where the dynamo is located, vs time to unravel physics of ``engine'' driving solar variability. -Track, for first time, solar active regions/solar storm regions simultaneously above and below solar surface to understand development and triggering of explosive events (Flares, Coronal Mass Ejections). Flight of Solar Sentinels, launch before and around next solar max to provide global view of the heliosphere, track solar active regions over entire sun, including for the first time the backside of Sun and describe the evolution of eruptions and flares from the Sun to Earth to Mars and beyond. Flight of Magnetospheric and Ionospheric Storm Probes, launch around solar max, geospace ``buoys'' in critical regions of Earth's space environment to determine its global and local response to solar variability. 2) A special data analysis and modeling program (Targeted Research & Technology) targeted at (1) obtaining a Sun-Earth systems model, (2) improving knowledge of space environmental conditions and variations over the solar cycle, (3) developing techniques and models for predicting solar/geospace disturbances which affect human technology, and (4) assimilating data from networks of spacecraft. 3) Orbital Technology Testbeds (Space Environment Testbeds) for low cost validation of rad-hard and rad-tolerant systems in high radiation orbits. - Implemented as missions of opportunity via cost-sharing partnerships with various international and national partners 4) Establishing and expanding partnerships for interdisciplinary science and applications with other NASA programs (Earth Science, Human Space Flight, Life Sciences), with other federal agencies (via interagency for National Space Weather Program, NOAA, NSF, DOE), international partners, academic community, and industry to obtain synergism and address common needs.

  18. Hot-electron generation by ``cavitating'' Langmuir turbulence in the nonlinear stage of the two-plasmon-decay instability

    NASA Astrophysics Data System (ADS)

    Vu, H. X.; DuBois, D. F.; Russell, D. A.; Myatt, J. F.

    2012-10-01

    The kinetic reduced-description particle-in-cell simulation technique has been applied to study the nonlinear stage of two-plasmon-decay (TPD) instability in an inhomogeneous plasma driven by crossed laser beams. The TPD instability is found to be a prolific generator of "cavitating" Langmuir turbulence. Langmuir "cavitons"—localized longitudinal electric fields, oscillating near the local electron plasma frequency, trapped in ponderomotive density depressions—collapse to dimensions of a few electron Debye lengths, where the electric field energy is collisionlessly transferred to electron kinetic energy. The resulting hot electrons can attain instantaneous temperatures up to 100 keV with net suprathermal heat flux out of the system of up to a few percent of the input laser energy. Scaling laws for this hot-electron generation by TPD, in regimes motivated by recent experiments on the Omega laser, were presented recently by Vu et al. (H. X. Vu, D. F. DuBois, D. A. Russell, and J. F. Myatt, Phys. Plasmas 19, 102703 (2012)). This paper concentrates on the microscopic mechanisms for hot-electron generation. The spatial distribution of the maxima of the electric field envelope modulus is found to be very spiky, with the distribution of electric field envelope maxima obeying Gaussian statistics. The cavitons are produced in density-depletion trenches produced by the combined ponderomotive interference of the crossed laser beams and the ponderomotive beats of the primary backward-going TPD Langmuir waves (LWs) resulting from the crossed beams. The Langmuir turbulence is strongest in the electron-density region near 0.241× the laser's critical density, where the forward LWs from the crossed-beam TPD are degenerate. Nucleation of cavitons is assisted by the modulation of the electron density in the trenches, which in turn is caused by the beating of the common forward-going LW and the pair of backward-going LWs. The autocorrelation function of the LW envelope field provides a near-universal shape for intense cavitons—in the neighborhood of the local field maxima. The hot-electron temperature is found to be approximately a linear function of the "caviton temperature" determined from the Gaussian distribution of caviton maxima. These diagnostics provide strong evidence for the importance of Langmuir caviton collapse in the generation of hot electrons by TPD. Extended Zakharov model predictions for TPD exhibit the same qualitative phenomena.

  19. Considering the Influence of Nonadaptive Evolution on Primate Color Vision.

    PubMed

    Jacobs, Rachel L; Bradley, Brenda J

    2016-01-01

    Color vision in primates is variable across species, and it represents a rare trait in which the genetic mechanisms underlying phenotypic variation are fairly well-understood. Research on primate color vision has largely focused on adaptive explanations for observed variation, but it remains unclear why some species have trichromatic or polymorphic color vision while others are red-green color blind. Lemurs, in particular, are highly variable. While some species are polymorphic, many closely-related species are strictly dichromatic. We provide the first characterization of color vision in a wild population of red-bellied lemurs (Eulemur rubriventer, Ranomafana National Park, Madagascar) with a sample size (87 individuals; NX chromosomes = 134) large enough to detect even rare variants (0.95 probability of detection at ≥ 3% frequency). By sequencing exon 5 of the X-linked opsin gene we identified opsin spectral sensitivity based on known diagnostic sites and found this population to be dichromatic and monomorphic for a long wavelength allele. Apparent fixation of this long allele is in contrast to previously published accounts of Eulemur species, which exhibit either polymorphic color vision or only the medium wavelength opsin. This unexpected result may represent loss of color vision variation, which could occur through selective processes and/or genetic drift (e.g., genetic bottleneck). To indirectly assess the latter scenario, we genotyped 55 adult red-bellied lemurs at seven variable microsatellite loci and used heterozygosity excess and M-ratio tests to assess if this population may have experienced a recent genetic bottleneck. Results of heterozygosity excess but not M-ratio tests suggest a bottleneck might have occurred in this red-bellied lemur population. Therefore, while selection may also play a role, the unique color vision observed in this population might have been influenced by a recent genetic bottleneck. These results emphasize the need to consider adaptive and nonadaptive mechanisms of color vision evolution in primates. PMID:26959829

  20. Functional and Anatomic Consequences of Subretinal Dosing in the Cynomolgus Macaque

    PubMed Central

    Nork, T. Michael; Murphy, Christopher J.; Kim, Charlene B. Y.; Hoeve, James N. Ver; Rasmussen, Carol A.; Miller, Paul E.; Wabers, Hugh D.; Neider, Michael W.; Dubielzig, Richard R.; McCulloh, Ryan J.; Christian, Brian J.

    2011-01-01

    Objectives To characterize functional and anatomic sequelae of a bleb induced by subretinal injection. Methods Subretinal injections (100 μl) of balance salt solution (BSS) were placed in the superotemporal macula of one eye in 3 cynomolgus macaques. Fellow eyes received intravitreal injections (100 μl) of BSS. Fundus photography, ocular coherence tomography (OCT) and multifocal electroretinography (mfERG) were obtained before and immediately after injection and again at intervals up to 3 months post injection. Histopathologic analyses included transmission electron microscopy (TEM) and immunohistochemistry (IHC) for glial fibrillary acidic protein (GFAP), rhodopsin, M/L-cone opsin and S-cone opsin. Results Retinas were re-attached by 2 days post-injection (by OCT). mfERG was suppressed post-subretinal injection within the subretinal injection bleb and surprisingly, also in regions far peripheral to this region. mfERG amplitudes were nearly completely recovered by 90 days. The spectral domain (SD)-OCT inner segment/outer segment (IS/OS) line had decreased reflectivity at 92 days. GFAP and S-cone staining were unaffected. Rhodopsin and M/L-cone opsins were partially displaced into the inner segments. TEM revealed disorganization of the outer segment rod (but not cone) disks. At all post-injection intervals, eyes with intravitreal injection were similar to baseline. Conclusions Subretinal injection is a promising route for drug delivery to the eye. Three months post subretinal injection, retinal function was nearly recovered, although reorganization of the outer segment rod disk remained disrupted. Understanding the functional and anatomic effects of subretinal injection per se is important for interpretation of the effects of compounds delivered to the subretinal space. Clinical relevance Subretinal injection is a new potential route for drug delivery to the eye. Separating drug effects from the procedural effects per se is critical. PMID:21911651

  1. Immuno-Histochemical Analysis of Rod and Cone Reaction to RPE65 Deficiency in the Inferior and Superior Canine Retina

    PubMed Central

    Klein, Daniela; Mendes-Madeira, Alexandra; Schlegel, Patrice; Rolling, Fabienne; Lorenz, Birgit; Haverkamp, Silke; Stieger, Knut

    2014-01-01

    Mutations in the RPE65 gene are associated with autosomal recessive early onset severe retinal dystrophy. Morphological and functional studies indicate early and dramatic loss of rod photoreceptors and early loss of S-cone function, while L and M cones remain initially functional. The Swedish Briard dog is a naturally occurring animal model for this disease. Detailed information about rod and cone reaction to RPE65 deficiency in this model with regard to their location within the retina remains limited. The aim of this study was to analyze morphological parameters of cone and rod viability in young adult RPE65 deficient dogs in different parts of the retina in order to shed light on local disparities in this disease. In retinae of affected dogs, sprouting of rod bipolar cell dendrites and horizontal cell processes was dramatically increased in the inferior peripheral part of affected retinae, while central inferior and both superior parts did not display significantly increased sprouting. This observation was correlated with photoreceptor cell layer thickness. Interestingly, while L/M cone opsin expression was uniformly reduced both in the superior and inferior part of the retina, S-cone opsin expression loss was less severe in the inferior part of the retina. In summary, in retinae of young adult RPE65 deficient dogs, the degree of rod bipolar and horizontal cell sprouting as well as of S-cone opsin expression depends on the location. As the human retinal pigment epithelium (RPE) is pigmented similar to the RPE in the inferior part of the canine retina, and the kinetics of photoreceptor degeneration in humans seems to be similar to what has been observed in the inferior peripheral retina in dogs, this area should be studied in future gene therapy experiments in this model. PMID:24466015

  2. Power analysis of tests for loss of selective constraint in cave crayfish and nonphotosynthetic plant lineages.

    PubMed

    Leebens-Mack, Jim; DePamphilis, Claude

    2002-08-01

    Loss of selective constraint on a gene may be expected following changes in the environment or life history that render its function unnecessary. The long-term persistence of protein-coding genes after the loss of known functional necessity can occur by chance or because of selective maintenance of an unknown gene function. The selective maintenance of an alternative gene function is not demonstrated by the failure of statistical tests to reject the hypothesis that there has been no change in the degree of constraint on the evolution of coding genes. Maintenance may be inferred, however, when power analyses of such tests demonstrate that there has been a sufficient number of nucleotide substitutions to detect the loss of selective constraint. Here, we describe a power analysis for tests of loss of constraint on protein-coding genes. The power analysis was applied to loss-of-constraint tests for opsin gene evolution in cave-dwelling crayfish and rbcL evolution in nonphotosynthetic parasitic plants. The power of previously applied tests for loss of constraint on cave crayfish opsin genes was insufficient to distinguish between chance retention and selective maintenance of opsin genes. However, the power of codon-based likelihood ratio tests for change in d(N)/d(S) (=omega) (nonsynonymous to synonymous change) did have sufficient power to detect a loss of constraint on rbcL associated with a loss of photosynthesis in most examples but failed to detect such a change in three independent lineages. We conclude that rbcL has been selectively maintained in these holoparasitic plant lineages. This conclusion suggests that either these taxa are photosynthetic for at least a part of their life or rbcL may have an unknown function in these plants unrelated to photosynthesis. PMID:12140241

  3. Inner retinal photoreceptors (IRPs) in mammals and teleost fish.

    PubMed

    Foster, Russell G; Bellingham, James

    2004-06-01

    Research over the past decade has provided overwhelming evidence that photoreception in the vertebrate eye is not confined to the rods and cones. The discovery of non-rod, non-cone ocular photoreceptors in mammals and fish arose from quite different lines of investigation. In transgenic mice entirely lacking functional rod and cone photoreceptors a range of responses to light, including the regulation of the circadian system and a pupillary light reflex, are preserved. Electrophysiological and imaging approaches were then able to characterise a coupled plexus of directly light sensitive ganglion cells. Most recently action spectroscopy has shown that a novel 'blue-light' sensitive photopigment based upon opsin/vitamin A (OP480) mediates these responses to light. Several candidate genes have emerged for OP480, with melanopsin being by far the strongest. A definitive link, however, between this gene and OP480 has still to be established. In contrast to the mammals, the discovery of inner retinal photoreceptors (IRPs) in fish started with the discovery of a new gene family (VA opsin). The teleost VA opsins form functional photopigments and are expressed in several different types of inner retinal neuron, including retinal horizontal cells. Recent studies have investigated the electrical properties of these photosensitive neurones, but their light-sensing role remains a matter of speculation. Thus the study of IRP is developing along quite separate lines. In the mammals the research is directed towards a molecular identification of the photopigment (OP480) and its cascade, whilst in fish the major effort is directed towards identifying a role for these novel photoreceptors using physiological approaches. The discovery of IRPs in the vertebrates tells us that despite 150 years of research, we still have much to learn about how the eye processes light. PMID:15170494

  4. Homologs of vertebrate Opn3 potentially serve as a light sensor in nonphotoreceptive tissue

    PubMed Central

    Koyanagi, Mitsumasa; Takada, Eiichiro; Nagata, Takashi; Tsukamoto, Hisao; Terakita, Akihisa

    2013-01-01

    Most opsins selectively bind 11-cis retinal as a chromophore to form a photosensitive pigment, which underlies various physiological functions, such as vision and circadian photoentrainment. Recently, opsin 3 (Opn3), originally called encephalopsin or panopsin, and its homologs were identified in various tissues including brain, eye, and liver in both vertebrates and invertebrates, including human. Because Opn3s are mainly expressed in tissues that are not considered to contain sufficient amounts of 11-cis retinal to form pigments, the photopigment formation ability of Opn3 has been of interest. Here, we report the successful expression of Opn3 homologs, pufferfish teleost multiple tissue opsin (PufTMT) and mosquito Opn3 (MosOpn3) and show that these proteins formed functional photopigments with 11-cis and 9-cis retinals. The PufTMT- and MosOpn3-based pigments have absorption maxima in the blue-to-green region and exhibit a bistable nature. These Opn3 homolog-based pigments activate Gi-type and Go-type G proteins light dependently, indicating that they potentially serve as light-sensitive Gi/Go-coupled receptors. We also demonstrated that mammalian cultured cells transfected with the MosOpn3 or PufTMT became light sensitive without the addition of 11-cis retinal and the photosensitivity retained after the continuous light exposure, showing a reusable pigment formation with retinal endogenously contained in culture medium. Interestingly, we found that the MosOpn3 also acts as a light sensor when constituted with 13-cis retinal, a ubiquitously present retinal isomer. Our findings suggest that homologs of vertebrate Opn3 might function as photoreceptors in various tissues; furthermore, these Opn3s, particularly the mosquito homolog, could provide a promising optogenetic tool for regulating cAMP-related G protein-coupled receptor signalings. PMID:23479626

  5. On the origins of arrestin and rhodopsin

    PubMed Central

    2008-01-01

    Background G protein coupled receptors (GPCRs) are the most numerous proteins in mammalian genomes, and the most common targets of clinical drugs. However, their evolution remains enigmatic. GPCRs are intimately associated with trimeric G proteins, G protein receptor kinases, and arrestins. We conducted phylogenetic studies to reconstruct the history of arrestins. Those findings, in turn, led us to investigate the origin of the photosensory GPCR rhodopsin. Results We found that the arrestin clan is comprised of the Spo0M protein family in archaea and bacteria, and the arrestin and Vps26 families in eukaryotes. The previously known animal arrestins are members of the visual/beta subfamily, which branched from the founding "alpha" arrestins relatively recently. Curiously, we identified both the oldest visual/beta arrestin and opsin genes in Cnidaria (but not in sponges). The arrestin clan has 14 human members: 6 alphas, 4 visual/betas, and 4 Vps26 genes. Others recently showed that the 3D structure of mammalian Vps26 and the biochemical function of the yeast alpha arrestin PalF are similar to those of beta arrestins. We note that only alpha arrestins have PY motifs (known to bind WW domains) in their C-terminal tails, and only visual/betas have helix I in the Arrestin N domain. Conclusion We identified ciliary opsins in Cnidaria and propose this subfamily is ancestral to all previously known animal opsins. That finding is consistent with Darwin's theory that eyes evolved once, and lends some support to Parker's hypothesis that vision triggered the Cambrian explosion of life forms. Our arrestin findings have implications on the evolution of GPCR signaling, and on the biological roles of human alpha arrestins. PMID:18664266

  6. Stable transgenesis in the marine annelid Platynereis dumerilii sheds new light on photoreceptor evolution

    PubMed Central

    Backfisch, Benjamin; Veedin Rajan, Vinoth Babu; Fischer, Ruth M.; Lohs, Claudia; Arboleda, Enrique; Tessmar-Raible, Kristin; Raible, Florian

    2013-01-01

    Research in eye evolution has mostly focused on eyes residing in the head. In contrast, noncephalic light sensors are far less understood and rather regarded as evolutionary innovations. We established stable transgenesis in the annelid Platynereis, a reference species for evolutionary and developmental comparisons. EGFP controlled by cis-regulatory elements of r-opsin, a characteristic marker for rhabdomeric photoreceptors, faithfully recapitulates known r-opsin expression in the adult eyes, and marks a pair of pigment-associated frontolateral eyelets in the brain. Unexpectedly, transgenic animals revealed an additional series of photoreceptors in the ventral nerve cord as well as photoreceptors that are located in each pair of the segmental dorsal appendages (notopodia) and project into the ventral nerve cord. Consistent with a photosensory function of these noncephalic cells, decapitated animals display a clear photoavoidance response. Molecular analysis of the receptors suggests that they differentiate independent of pax6, a gene involved in early eye development of many metazoans, and that the ventral cells may share origins with the Hesse organs in the amphioxus neural tube. Finally, expression analysis of opn4×-2 and opn4m-2, two zebrafish orthologs of Platynereis r-opsin, reveals that these genes share expression in the neuromasts, known mechanoreceptors of the lateral line peripheral nervous system. Together, this establishes that noncephalic photoreceptors are more widespread than assumed, and may even reflect more ancient aspects of sensory systems. Our study marks significant advance for the understanding of photoreceptor cell (PRC) evolution and development and for Platynereis as a functional lophotrochozoan model system. PMID:23284166

  7. From Blue Light to Clock Genes in Zebrafish ZEM-2S Cells

    PubMed Central

    Ramos, Bruno C. R.; Moraes, Maria Nathália C. M.; Poletini, Maristela O.; Lima, Leonardo H. R. G.; Castrucci, Ana Maria L.

    2014-01-01

    Melanopsin has been implicated in the mammalian photoentrainment by blue light. This photopigment, which maximally absorbs light at wavelengths between 470 and 480 nm depending on the species, is found in the retina of all classes of vertebrates so far studied. In mammals, melanopsin activation triggers a signaling pathway which resets the circadian clock in the suprachiasmatic nucleus (SCN). Unlike mammals, Drosophila melanogaster and Danio rerio do not rely only on their eyes to perceive light, in fact their whole body may be capable of detecting light and entraining their circadian clock. Melanopsin, teleost multiple tissue (tmt) opsin and others such as neuropsin and va-opsin, are found in the peripheral tissues of Danio rerio, however, there are limited data concerning the photopigment/s or the signaling pathway/s directly involved in light detection. Here, we demonstrate that melanopsin is a strong candidate to mediate synchronization of zebrafish cells. The deduced amino acid sequence of melanopsin, although being a vertebrate opsin, is more similar to invertebrate than vertebrate photopigments, and melanopsin photostimulation triggers the phosphoinositide pathway through activation of a Gq/11-type G protein. We stimulated cultured ZEM-2S cells with blue light at wavelengths consistent with melanopsin maximal absorption, and evaluated the time course expression of per1b, cry1b, per2 and cry1a. Using quantitative PCR, we showed that blue light is capable of slightly modulating per1b and cry1b genes, and drastically increasing per2 and cry1a expression. Pharmacological assays indicated that per2 and cry1a responses to blue light are evoked through the activation of the phosphoinositide pathway, which crosstalks with nitric oxide (NO) and mitogen activated protein MAP kinase (MAPK) to activate the clock genes. Our results suggest that melanopsin may be important in mediating the photoresponse in Danio rerio ZEM-2S cells, and provide new insights about the modulation of clock genes in peripheral clocks. PMID:25184495

  8. Quantum mechanical/molecular mechanical structure, enantioselectivity, and spectroscopy of hydroxyretinals and insights into the evolution of color vision in small white butterflies.

    PubMed

    Sekharan, Sivakumar; Yokoyama, Shozo; Morokuma, Keiji

    2011-12-29

    Since Vogt's discovery of A(3)-retinal or 3-hydroxyretinal in insects in 1983 and Matsui's discovery of A(4)-retinal or 4-hydroxyretinal in firefly squid in 1988, hydroxyretinal-protein interactions mediating vision have remained largely unexplored. In the present study, A(3)- and A(4)-retinals are theoretically incorporated into squid and bovine visual pigments by use of the hybrid quantum mechanics/molecular mechanics [SORCI+Q//B3LYP/6-31G(d):Amber96] method, and insights into structure, enantioselectivity, and spectroscopy are gathered and presented for the first time. Contrary to general perception, our findings rule out the formation of a hydrogen bond between the hydroxyl-bearing β-ionone ring portion of retinal and opsin. Compared to A(1)-pigments, A(3)- and A(4)-pigments exhibit slightly blue-shifted absorption maxima due to increase in bond-length alternation of the hydroxyretinal. We suggest that (i) the binding site of firefly squid (Watasenia scintillans) opsin is very similar to that of the Japanese common squid (Todarodes pacificus) opsin; (ii) the molecular mechanism of spectral tuning in small white butterflies involve sites S116 and T185 and breaking of a hydrogen bond between sites E180 and T185; and finally (iii) A(3)-retinal may have occurred during the conversion of A(1)- to A(2)-retinal and insects may have acquired them, in order to absorb light in the blue-green wavelength region and to speed up the G-protein signaling cascade. PMID:22087641

  9. Gene Therapy Rescues Cone Structure and Function in the 3-Month-Old rd12 Mouse: A Model for Midcourse RPE65 Leber Congenital Amaurosis

    PubMed Central

    Li, Xia; Li, Wensheng; Dai, Xufeng; Kong, Fansheng; Zheng, Qinxiang; Zhou, Xiangtian; Lü, Fan; Chang, Bo; Rohrer, Bärbel; Hauswirth, William. W.; Qu, Jia; Pang, Ji-jing

    2011-01-01

    Purpose. RPE65 function is necessary in the retinal pigment epithelium (RPE) to generate chromophore for all opsins. Its absence results in vision loss and rapid cone degeneration. Recent Leber congenital amaurosis type 2 (LCA with RPE65 mutations) phase I clinical trials demonstrated restoration of vision on RPE65 gene transfer into RPE cells overlying cones. In the rd12 mouse, a naturally occurring model of RPE65-LCA early cone degeneration was observed; however, some peripheral M-cones remained. A prior study showed that AAV-mediated RPE65 expression can prevent early cone degeneration. The present study was conducted to test whether the remaining cones in older rd12 mice can be rescued. Methods. Subretinal treatment with the scAAV5-smCBA-hRPE65 vector was initiated at postnatal day (P)14 and P90. After 2 months, electroretinograms were recorded, and cone morphology was analyzed by using cone-specific peanut agglutinin and cone opsin–specific antibodies. Results. Cone degeneration started centrally and spread ventrally, with cells losing cone-opsin staining before that for the PNA-lectin–positive cone sheath. Gene therapy starting at P14 resulted in almost wild-type M- and S-cone function and morphology. Delaying gene-replacement rescued the remaining M-cones, and most important, more M-cone opsin–positive cells were identified than were present at the onset of gene therapy, suggesting that opsin expression could be reinitiated in cells with cone sheaths. Conclusions. The results support and extend those of the previous study that gene therapy can stop early cone degeneration, and, more important, they provide proof that delayed treatment can restore the function and morphology of the remaining cones. These results have important implications for the ongoing LCA2 clinical trials. PMID:21169527

  10. Considering the Influence of Nonadaptive Evolution on Primate Color Vision

    PubMed Central

    Jacobs, Rachel L.; Bradley, Brenda J.

    2016-01-01

    Color vision in primates is variable across species, and it represents a rare trait in which the genetic mechanisms underlying phenotypic variation are fairly well-understood. Research on primate color vision has largely focused on adaptive explanations for observed variation, but it remains unclear why some species have trichromatic or polymorphic color vision while others are red-green color blind. Lemurs, in particular, are highly variable. While some species are polymorphic, many closely-related species are strictly dichromatic. We provide the first characterization of color vision in a wild population of red-bellied lemurs (Eulemur rubriventer, Ranomafana National Park, Madagascar) with a sample size (87 individuals; NX chromosomes = 134) large enough to detect even rare variants (0.95 probability of detection at ≥ 3% frequency). By sequencing exon 5 of the X-linked opsin gene we identified opsin spectral sensitivity based on known diagnostic sites and found this population to be dichromatic and monomorphic for a long wavelength allele. Apparent fixation of this long allele is in contrast to previously published accounts of Eulemur species, which exhibit either polymorphic color vision or only the medium wavelength opsin. This unexpected result may represent loss of color vision variation, which could occur through selective processes and/or genetic drift (e.g., genetic bottleneck). To indirectly assess the latter scenario, we genotyped 55 adult red-bellied lemurs at seven variable microsatellite loci and used heterozygosity excess and M-ratio tests to assess if this population may have experienced a recent genetic bottleneck. Results of heterozygosity excess but not M-ratio tests suggest a bottleneck might have occurred in this red-bellied lemur population. Therefore, while selection may also play a role, the unique color vision observed in this population might have been influenced by a recent genetic bottleneck. These results emphasize the need to consider adaptive and nonadaptive mechanisms of color vision evolution in primates. PMID:26959829

  11. The ecoresponsive genome of Daphnia pulex

    SciTech Connect

    Colbourne, John K.; Pfrender, Michael E.; Gilbert, Donald; Thomas, W. Kelley; Tucker, Abraham; Oakley, Todd H.; Tokishita, Shinichi; Aerts, Andrea; Arnold, Georg J.; Basu, Malay Kumar; Bauer, Darren J.; Caceres, Carla E.; Carmel, Liran; Casola, Claudio; Choi, Jeong-Hyeon; Detter, John C.; Dong, Qunfeng; Dusheyko, Serge; Eads, Brian D.; Frohlich, Thomas; Geiler-Samerotte, Kerry A.; Gerlach, Daniel; Hatcher, Phil; Jogdeo, Sanjuro; Krijgsveld, Jeroen; Kriventseva, Evgenia V; Kültz, Dietmar; Laforsch, Christian; Lindquist, Erika; Lopez, Jacqueline; Manak, Robert; Muller, Jean; Pangilinan, Jasmyn; Patwardhan, Rupali P.; Pitluck, Samuel; Pritham, Ellen J.; Rechtsteiner, Andreas; Rho, Mina; Rogozin, Igor B.; Sakarya, Onur; Salamov, Asaf; Schaack, Sarah; Shapiro, Harris; Shiga, Yasuhiro; Skalitzky, Courtney; Smith, Zachary; Souvorov, Alexander; Sung, Way; Tang, Zuojian; Tsuchiya, Dai; Tu, Hank; Vos, Harmjan; Wang, Mei; Wolf, Yuri I.; Yamagata, Hideo; Yamada, Takuji; Ye, Yuzhen; Shaw, Joseph R.; Andrews, Justen; Crease, Teresa J.; Tang, Haixu; Lucas, Susan M.; Robertson, Hugh M.; Bork, Peer; Koonin, Eugene V.; Zdobnov, Evgeny M.; Grigoriev, Igor V.; Lynch, Michael; Boore, Jeffrey L.

    2011-02-04

    This document provides supporting material related to the sequencing of the ecoresponsive genome of Daphnia pulex. This material includes information on materials and methods and supporting text, as well as supplemental figures, tables, and references. The coverage of materials and methods addresses genome sequence, assembly, and mapping to chromosomes, gene inventory, attributes of a compact genome, the origin and preservation of Daphnia pulex genes, implications of Daphnia's genome structure, evolutionary diversification of duplicated genes, functional significance of expanded gene families, and ecoresponsive genes. Supporting text covers chromosome studies, gene homology among Daphnia genomes, micro-RNA and transposable elements and the 46 Daphnia pulex opsins. 36 figures, 50 tables, 183 references.

  12. Seeing the light: a photonic visual prosthesis for the blind

    NASA Astrophysics Data System (ADS)

    Degenaar, Patrick; Grossman, Nir; McGovern, Brian; Neil, Mark; Drakakis, Emmanuel; Nikolic, Konstantin

    2009-02-01

    This paper highlights how the genetic incorporation of artificial opsins into the retina can lead to a new class of retinal prosthesis. We demonstrate the efficacy of incorporating channelrhodopsin into neuron cells in-vitro and show how that can be scaled to in-vivo. We show that we need typically 100mW/cm2 of instantaneous light intensity on the neuron in order to stimulate action potentials which results in 10W/cm2 required from the light source. We thus use GaN LED arrays to provide spatially controlled stimulation which is of sufficient brightness to stimulate the cells.

  13. Optobionic vision—a new genetically enhanced light on retinal prosthesis

    NASA Astrophysics Data System (ADS)

    Degenaar, Patrick; Grossman, Nir; Memon, Muhammad Ali; Burrone, Juan; Dawson, Martin; Drakakis, Emmanuel; Neil, Mark; Nikolic, Konstantin

    2009-06-01

    The recent discovery that neurons can be photostimulated via genetic incorporation of artificial opsins is creating a revolution in the field of neural stimulation. In this paper we show its potential in the field of retinal prosthesis. We show that we need typically 100 mW cm-2 in instantaneous light intensity on the neuron in order to stimulate action potentials. We also show how this can be reduced down to safe levels in order to negate thermal and photochromic damage to the eye. We also describe a gallium nitride LED light source which is also able to generate patterns of the required intensity in order to transfer reliable images.

  14. Circuit-breakers: optical technologies for probing neural signals and systems.

    PubMed

    Zhang, Feng; Aravanis, Alexander M; Adamantidis, Antoine; de Lecea, Luis; Deisseroth, Karl

    2007-08-01

    Neuropsychiatric disorders, which arise from a combination of genetic, epigenetic and environmental influences, epitomize the challenges faced in understanding the mammalian brain. Elucidation and treatment of these diseases will benefit from understanding how specific brain cell types are interconnected and signal in neural circuits. Newly developed neuroengineering tools based on two microbial opsins, channelrhodopsin-2 (ChR2) and halorhodopsin (NpHR), enable the investigation of neural circuit function with cell-type-specific, temporally accurate and reversible neuromodulation. These tools could lead to the development of precise neuromodulation technologies for animal models of disease and clinical neuropsychiatry. PMID:17643087

  15. Optobionic vision--a new genetically enhanced light on retinal prosthesis.

    PubMed

    Degenaar, Patrick; Grossman, Nir; Memon, Muhammad Ali; Burrone, Juan; Dawson, Martin; Drakakis, Emmanuel; Neil, Mark; Nikolic, Konstantin

    2009-06-01

    The recent discovery that neurons can be photostimulated via genetic incorporation of artificial opsins is creating a revolution in the field of neural stimulation. In this paper we show its potential in the field of retinal prosthesis. We show that we need typically 100 mW cm(-2) in instantaneous light intensity on the neuron in order to stimulate action potentials. We also show how this can be reduced down to safe levels in order to negate thermal and photochromic damage to the eye. We also describe a gallium nitride LED light source which is also able to generate patterns of the required intensity in order to transfer reliable images. PMID:19458396

  16. Cone photopigment variations in Cebus apella monkeys evidenced by electroretinogram measurements and genetic analysis

    PubMed Central

    Soares, Juliana G.M.; Fiorani, Mario; Araujo, Eduardo A.; Zana, Yossi; Bonci, Daniela M.O.; Neitz, Maureen; Ventura, Dora F.; Gattass, Ricardo

    2011-01-01

    We investigated the color vision pattern in male and female Cebus apella monkeys by means of electroretinogram measurements and genetic analysis. Our objective was to establish a simple, fast and efficient protocol in order to determine the chromatic vision pattern in Cebus monkeys. We found five among ten possible different phenotypes, two trichromats and three dichromats. We also found that Cebus present a new allele with spectral peak near 552 nm, with the amino acid combination SFT at positions 180, 277 and 285 of the opsin gene, in addition to the previously described SYT, AFT and AFA alleles. PMID:19883678

  17. Solar EUV Variability from FISM and SDO/EVE During Solar Minimum, Active, and Flaring Time Periods

    NASA Technical Reports Server (NTRS)

    Chamberlin, Phillip C.

    2011-01-01

    The Living With a Star (LWS) Focus Science Team has identified three periods of different solar activity levels for which they will be determining the Earth's Ionosphere and Thermosphere response. Not only will the team be comparing individual models (e.g. FLIP, T1MEGCM, GLOW) outcome driven by the various levels of solar activity, but the models themselves will also be compared. These models all rely on the input solar EUV (0.1 -190 nm) irradiance to drive the variability. The Flare Irradiance Spectral Model (FISM) and the EUV Variability Experiment (EVE) onboard provide the Solar Dynamics Observatory (SDO) provide the most accurate quantification of these irradiances. Presented and discussed are how much the solar EUV irradiance changes during these three scenarios, both as a function of activity and wavelength.

  18. Cryogenic mechanisms for scanning and interchange of the Fabry-Perot interferometers in the ISO long wavelength spectrometer

    NASA Technical Reports Server (NTRS)

    Davis, G. R.; Furniss, I.; Patrick, T. J.; Sidey, R. C.; Towlson, W. A.

    1991-01-01

    The Infrared Space Observatory (ISO) is an ESA cornerstone mission for infrared astronomy. Schedules for launch in 1993, its four scientific instruments will provide unprecedented sensitivity and spectral resolution at wavelengths which are inaccessible using ground-based techniques. One of these, the Long Wavelength Spectrometer (LWS), will operate in the 45 to 180 micron region (Emery et. al., 1985) and features two Fabry-Perot interferometers mounted on an interchange mechanism. The entire payload module of the spacecraft, comprising the 60 cm telescope and the four focal plane instruments, is maintained at 2 to 4 K by an onboard supply of liquid helium. The mechanical design and testing of the cryogenic interferometer and interchange mechanisms are described.

  19. A Physical Model of the Radiation Belt

    NASA Astrophysics Data System (ADS)

    Fok, M.

    2006-12-01

    A data-driven physical model of the radiation belt has been activated, based on the kinetic model previously developed by Fok and coworkers. This model calculates radiation belt electron fluxes from 10 keV to 4 MeV in the inner magnetosphere. The model takes into account realistic, time-varying magnetic field and considers effects of wave-particle interactions. The only inputs to the model are upstream solar wind and concurrent Dst data. With the real-time solar wind data supplied by NOAA and Dst data from the Kyoto University, the model is running in real time to provide now-casting of the radiation belt environment. In this paper, the model logic and formulation are discussed. The model results for several storm simulations are presented. The paper concludes with discussion of further improvements of the model to better achieve additional LWS goals.

  20. Resolving The Azimuthal Ambiguity In Vector Magnetograms Away From Disk Centre With The Solenoidal Condition

    NASA Astrophysics Data System (ADS)

    Crouch, Ashley D.; Barnes, G.

    2007-05-01

    We employ the divergence-free property of magnetic fields to resolve the azimuthal ambiguity in solar vector magnetograms. We show that the ambiguity can be resolved away from disk centre if one knows the line-of-sight derivative of the magnetic field components in the directions parallel and transverse to the line-of-sight. However, knowing only the line-of-sight derivative of the line-of-sight component of the magnetic field is not sufficient except at disk centre. Thus, multi-height vector magnetogram data can be used to resolve the ambiguity provided that all the line-of-sight derivatives can be determined reliably. We use a simple theoretical example, consisting of two submerged magnetic point sources, to demonstrate our findings. This work was supported by funding from NASA/LWS under contract NNH05CC75C.

  1. Asteroids as Calibration Standards in the Thermal Infrared -- Applications and Results from ISO

    NASA Astrophysics Data System (ADS)

    Müller, T. G.; Lagerros, J. S. V.

    Asteroids have been used extensively as calibration sources for ISO. We summarise the asteroid observational parameters in the thermal infrared and explain the important modelling aspects. Ten selected asteroids were extensively used for the absolute photometric calibration of ISOPHOT in the far-IR. Additionally, the point-like and bright asteroids turned out to be of great interest for many technical tests and calibration aspects. They have been used for testing the calibration for SWS and LWS, the validation of relative spectral response functions of different bands, for colour correction and filter leak tests. Currently, there is a strong emphasis on ISO cross-calibration, where the asteroids contribute in many fields. Well known asteroids have also been seen serendipitously in the CAM Parallel Mode and the PHT Serendipity Mode, allowing for validation and improvement of the photometric calibration of these special observing modes.

  2. 20th Century Solar Spectral Irradiance Modeling Based on Solar Cycle 22 and 23 Measurements

    NASA Astrophysics Data System (ADS)

    Morrill, J.; McMullin, D.; Floyd, L.; Newmark, J.

    2008-12-01

    Our long term solar spectral irradiance model under development will include EUV, UV, Visible, and IR spectral irradiance estimates. This model is based, in part, on Ca II K images collected from various observatories. For example, images from the Mt. Wilson Observatory supplemented with Greenwhich Sunspot data are used in the model to produce irradiance estimates for the time period before space-based spectral irradiance observations. The Mt. Wilson Ca II K film archive extending back to 1915 has been recently digitized. We expect to generate estimated spectra back to the start of this data set. We discuss the development details of the various spectral component derived from various spectra and proxies measured during Solar Cycle 22 and 23. Measured and estimated spectra will be compared in time series of various spectral bands as well as in spectra for days with specific solar surface configurations. This work is supported by the NASA LWS program.

  3. Progress Towards the Solar Dynamics Observatory

    NASA Astrophysics Data System (ADS)

    Thompson, B.; Schwer, K.

    The Solar Dynamics Observatory, or SDO, is scheduled to be the first mission to launch in 2007 under the new Living With a Star (LWS) program. It builds on the success of SOHO and other recent solar missions, but will observe the Sun at greater resolution and faster time cadence with a set of remote sensing instruments generating data in excess of 100 Mbps. The Science Definition Team produced a report consisting of a series of science objectives and a baseline instrument complement. Instrument proposals were due in April 2002, with selection to occur in the late summer of 2002. The spacecraft is being built at NASA Goddard Spacecraft Center by a team of engineers which are currently undergoing the formulation process. The presentation will discuss the current status of the science investigation selection and the spacecraft formulation.

  4. ISO Detections of C3 and NH in SGR B2

    NASA Astrophysics Data System (ADS)

    Goicoechea, J. R.; Cernicharo, J.; Caux, E.

    2000-11-01

    We present ISO/LWS Fabry-Perot data of the prominent Sagittarius B2 molecular cloud. Several ro-vibrational lines of triatomic carbon, C3, have been detected towards the central position of the cloud. Preliminary calculations indicate χ(C3)≃10-8. It is the first time that pure rotational lines of NH have been observed in the dense interstellar medium towards the galactic center. The abundance of NH is a few 10-9. We also report a tentative first detection of SH rotational lines. Polyatomic molecules will have a weak contribution from their pure rotational transitions to the far-infrared spectrum. However, we suggest that they could be, through their low-lying vibrational bending modes, the dominant carriers of emission/absorption in the spectrum of bright far-infrared sources.

  5. Development of a long wavelength spectrometer for the 24-channel multispectral scanner: Instructions for installation, start-up, and adjustment

    NASA Technical Reports Server (NTRS)

    1974-01-01

    The basic information is presented, which is required for start-up and operation of two long-wavelength focal-plane and cooler assemblies, including the amplifiers and temperature control systems. The focal plane systems, referred to as the long wavelength spectrometer (LWS) were developed for direct replacement of Arrays 3 and 4 into the multispectral scanner presently being operated by the NASA Manned Spacecraft Center Facility, and Laboratory Support Branch. The equipment is comprised of two major sub-assemblies: Array 3 with three indium antimonide detector channels and Array 4 with seven mercury doped Germanium detector channels. Each array is mounted on a cryogenic cooler and includes the vacuum housings, mounting hardware (x, y, z translation and rotation stages) and detector signal conditioning, temperature control and monitoring electronics. The two arrays were designed to operate independently and do not share common equipment (viz power supplies, housings, mounts, etc.).

  6. The Solar Dynamics Observatory and Its Contributions to Space Weather

    NASA Technical Reports Server (NTRS)

    Chamberlin, Phillip C.

    2011-01-01

    The Solar Dynamics Observatory (SDO) was launched on 11 February 2010 and has worked flawlessly in its first year and a half of operation. SDO was the first mission launched for NASA's Living With a Star Program (LWS), so its focus is not only studying the causes and drivers of the variable Sun, but also how these variations force similar changes in the Earth and other objects within the Heliosphere. Due to SDO's many Space Weather goals, this presentation will not only show some of the recent, ground-breaking new results provided by SDO, but also focus on the real-time Space Weather advances provided by this spacecraft. A main theme throughout this talk will be methods and tools that researchers around the world can utilize to access and manipulate the SDO data real-time for both fundamental science and Space Weather monitoring purposes.

  7. The NIRSPEC Data Reduction Pipeline for the Keck Observatory Archive

    NASA Astrophysics Data System (ADS)

    Tran, Hien D.; Cohen, R.; Mader, J. A.; Colson, A.; Berriman, G. Bruce; Gelino, Christopher R.; KOA Team

    2016-01-01

    The Keck Observatory Archive (KOA), a collaboration between the NASA Exoplanet Science Institute and the W. M. Keck Observatory, serves science and calibration data for all current and retired instruments from the twin Keck Telescopes. In addition to the raw data, we publicly serve quick-look, reduced data products for four instruments (HIRES, LWS, NIRC2 and OSIRIS), so that KOA users can easily assess the quality and scientific content of the data. In this paper we present the design and implementation of the data reduction pipeline (DRP) for the NIRSPEC instrument for use with KOA. We discuss the publicly available reduction packages for NIRSPEC, the challenges encountered when designing this fully automated DRP and the algorithm used to determine wavelength calibration from sky lines. The reduced data products from the NIRSPEC DRP are expected to be available in KOA by mid-2016.

  8. Long wavelength infrared camera (LWIRC): a 10 micron camera for the Keck Telescope

    SciTech Connect

    Wishnow, E.H.; Danchi, W.C.; Tuthill, P.; Wurtz, R.; Jernigan, J.G.; Arens, J.F.

    1998-05-01

    The Long Wavelength Infrared Camera (LWIRC) is a facility instrument for the Keck Observatory designed to operate at the f/25 forward Cassegrain focus of the Keck I telescope. The camera operates over the wavelength band 7-13 {micro}m using ZnSe transmissive optics. A set of filters, a circular variable filter (CVF), and a mid-infrared polarizer are available, as are three plate scales: 0.05``, 0.10``, 0.21`` per pixel. The camera focal plane array and optics are cooled using liquid helium. The system has been refurbished with a 128 x 128 pixel Si:As detector array. The electronics readout system used to clock the array is compatible with both the hardware and software of the other Keck infrared instruments NIRC and LWS. A new pre-amplifier/A-D converter has been designed and constructed which decreases greatly the system susceptibility to noise.

  9. Characterizing the Properties of Coronal Magnetic Null Points

    NASA Astrophysics Data System (ADS)

    Barnes, Graham; DeRosa, Marc; Wagner, Eric

    2015-08-01

    The topology of the coronal magnetic field plays a role in a wide range of phenomena, from Coronal Mass Ejections (CMEs) through heating of the corona. One fundamental topological feature is the null point, where the magnetic field vanishes. These points are natural sites of magnetic reconnection, and hence the release of energy stored in the magnetic field. We present preliminary results of a study using data from the Helioseismic and Magnetic Imager aboard NASA's Solar Dynamics Observatory to characterize the properties and evolution of null points in a Potential Field Source Surface model of the coronal field. The main properties considered are the lifetime of the null points, their distribution with height, and how they form and subsequently vanish.This work is supported by NASA/LWS Grant NNX14AD45G, and by NSF/SHINE grant 1357018.

  10. MHD modeling of the solar corona: Progress and challenges

    NASA Astrophysics Data System (ADS)

    Linker, Jon; Mikic, Zoran; Lionello, Roberto; Riley, Pete; Titov, Viacheslav; Torok, Tibor

    2012-07-01

    The Sun and its activity is the ultimate driver of space weather at Earth. This influence occurs not only via eruptive phenomena such as coronal mass ejections, but also through the structure of the corona itself, which forms the genesis of fast solar wind streams that trigger recurrent geomagnetic activity. Coronal structure also determines the connection of the ambient interplanetary magnetic field to CME-related shocks and impulsive solar flares, and thus controls where solar energetic particles propagate. In this talk we describe both the present state of the art and new directions in coronal modeling for both dynamic and slowly varying phenomena. We discuss the challenges to incorporating these capabilities into future space weather forecasting and specification models. Supported by NASA through the HTP, LWS, and SR&T programs, by NSF through the FESD and CISM programs, and by the AFOSR Space Science program.

  11. Expression of carnitine palmitoyl-CoA transferase-1B is influenced by a cis-acting eQTL in two chicken lines selected for high and low body weight.

    PubMed

    Ka, Sojeong; Markljung, Ellen; Ring, Henrik; Albert, Frank W; Harun-Or-Rashid, Mohammad; Wahlberg, Per; Garcia-Roves, Pablo M; Zierath, Juleen R; Denbow, D Michael; Pbo, Svante; Siegel, Paul B; Andersson, Leif; Hallbk, Finn

    2013-05-01

    Carnitine palmitoyl-CoA transferase-1B is a mitochondrial enzyme in the fatty acid oxidation pathway. In a previous study, CPT1B was identified as differentially expressed in the hypothalamus of two lines of chickens established by long-term selection for high (HWS) or low (LWS) body weight. Mammals have three paralogs (CPT1a, b and c) while nonmammalian vertebrates only have two (CPT1A, B). CPT1A is expressed in liver and CPT1B in muscle. CPT1c is expressed in hypothalamus, where it regulates feeding and energy expenditure. We identified an intronic length polymorphism, fixed for different alleles in the two populations, and mapped the hitherto missing CPT1B locus in the chicken genome assembly, to the distal tip of chromosome 1p. Based on molecular phylogeny and gene synteny we suggest that chicken CPT1B is pro-orthologous of the mammalian CPT1c. Chicken CPT1B was differentially expressed in both muscle and hypothalamus but in opposite directions: higher levels in hypothalamus but lower levels in muscle in the HWS than in the LWS line. Using an advanced intercross population of the lines, we found CPT1B expression to be influenced by a cis-acting expression quantitative trait locus in muscle. The increased expression in hypothalamus and reduced expression in muscle is consistent with an increased food intake in the HWS line and at the same time reduced fatty acid oxidation in muscle yielding a net accumulation of energy intake and storage. The altered expression of CPT1B in hypothalamus and peripheral tissue is likely to be a mechanism contributing to the remarkable difference between lines. PMID:23512741

  12. Hot-electron production and suprathermal heat flux scaling with laser intensity from the two-plasmon-decay instability

    NASA Astrophysics Data System (ADS)

    Vu, H. X.; DuBois, D. F.; Myatt, J. F.; Russell, D. A.

    2012-10-01

    The fully kinetic reduced-description particle-in-cell (RPIC) method has been applied to simulations of two-plasmon-decay (TPD) instability, driven by crossed laser beams, in an inhomogeneous plasma for parameters consistent with recent direct-drive experiments related to laser-driven inertial fusion. The nonlinear saturated state is characterized by very spiky electric fields, with Langmuir cavitation occurring preferentially inside density channels produced by the ponderomotive beating of the crossed laser beams and the primary TPD Langmuir waves (LWs). The heated electron distribution function is, in all cases, bi-Maxwellian, with instantaneous hot-electron temperatures in the range 60-100 keV. The net hot-electron energy flux out of the system is a small fraction (˜1% to 2%) of the input laser intensity in these simulations. Scalings of the hot-electron temperature and suprathermal heat flux as functions of the laser intensity are obtained numerically from RPIC simulations. These simulations lead to the preliminary conclusion that Langmuir cavitation and collapse provide dissipation by producing suprathermal electrons, which stabilize the system in saturation and drive the LW spectrum to the small dissipation scales at the Landau cutoff. The Langmuir turbulence originates at an electron density 0.241× the laser's critical density, where the crossed laser beams excite a "triad" mode—a common forward LW plus a pair of backward LWs. Remnants of this "triad" evolve in k-space and dominate the time-averaged energy spectrum. At times exceeding 10 ps, the excited Langmuir turbulence spreads toward lower densities. Comparisons of RPIC simulations with the extended Zakharov model are presented in appropriate regimes, and the necessary requirements for the validity of a quasi-linear Zakharov model (where the spatially averaged electron-velocity distribution is evolved) are verified by RPIC simulation results.

  13. The Far Infrared Lines of OH as Molecular Cloud Diagnostics

    NASA Technical Reports Server (NTRS)

    Smith, Howard A.

    2004-01-01

    Future IR missions should give some priority to high resolution spectroscopic observations of the set of far-IR transitions of OH. There are 15 far-IR lines arising between the lowest eight rotational levels of OH, and ISO detected nine of them. Furthermore, ISO found the OH lines, sometimes in emission and sometimes in absorption, in a wide variety of galactic and extragalactic objects ranging from AGB stars to molecular clouds to active galactic nuclei and ultra-luminous IR galaxies. The ISO/LWS Fabry-Perot resolved the 119 m doublet line in a few of the strong sources. This set of OH lines provides a uniquely important diagnostic for many reasons: the lines span a wide wavelength range (28.9 m to 163.2 m); the transitions have fast radiative rates; the abundance of the species is relatively high; the IR continuum plays an important role as a pump; the contribution from shocks is relatively minor; and, not least, the powerful centimeter-wave radiation from OH allows comparison with radio and VLBI datasets. The problem is that the large number of sensitive free parameters, and the large optical depths of the strongest lines, make modeling the full set a difficult job. The SWAS montecarlo radiative transfer code has been used to analyze the ISO/LWS spectra of a number of objects with good success, including in both the lines and the FIR continuum; the DUSTY radiative transfer code was used to insure a self-consistent continuum. Other far IR lines including those from H2O, CO, and [OI] are also in the code. The OH lines all show features which future FIR spectrometers should be able to resolve, and which will enable further refinements in the details of each cloud's structure. Some examples are given, including the case of S140, for which independent SWAS data found evidence for bulk flows.

  14. Hot-electron production and suprathermal heat flux scaling with laser intensity from the two-plasmon-decay instability

    SciTech Connect

    Vu, H. X.; DuBois, D. F.; Myatt, J. F.; Russell, D. A.

    2012-10-15

    The fully kinetic reduced-description particle-in-cell (RPIC) method has been applied to simulations of two-plasmon-decay (TPD) instability, driven by crossed laser beams, in an inhomogeneous plasma for parameters consistent with recent direct-drive experiments related to laser-driven inertial fusion. The nonlinear saturated state is characterized by very spiky electric fields, with Langmuir cavitation occurring preferentially inside density channels produced by the ponderomotive beating of the crossed laser beams and the primary TPD Langmuir waves (LWs). The heated electron distribution function is, in all cases, bi-Maxwellian, with instantaneous hot-electron temperatures in the range 60-100 keV. The net hot-electron energy flux out of the system is a small fraction ({approx}1% to 2%) of the input laser intensity in these simulations. Scalings of the hot-electron temperature and suprathermal heat flux as functions of the laser intensity are obtained numerically from RPIC simulations. These simulations lead to the preliminary conclusion that Langmuir cavitation and collapse provide dissipation by producing suprathermal electrons, which stabilize the system in saturation and drive the LW spectrum to the small dissipation scales at the Landau cutoff. The Langmuir turbulence originates at an electron density 0.241 Multiplication-Sign the laser's critical density, where the crossed laser beams excite a 'triad' mode-a common forward LW plus a pair of backward LWs. Remnants of this 'triad' evolve in k-space and dominate the time-averaged energy spectrum. At times exceeding 10 ps, the excited Langmuir turbulence spreads toward lower densities. Comparisons of RPIC simulations with the extended Zakharov model are presented in appropriate regimes, and the necessary requirements for the validity of a quasi-linear Zakharov model (where the spatially averaged electron-velocity distribution is evolved) are verified by RPIC simulation results.

  15. 90 GHz AND 150 GHz OBSERVATIONS OF THE ORION M42 REGION. A SUBMILLIMETER TO RADIO ANALYSIS

    SciTech Connect

    Dicker, S. R.; Korngut, P. M.; Devlin, M. J.; Mason, B. S.; Cotton, W. D.; Compiegne, M.; Martin, P. G.; Ade, P. A. R; Tucker, C.; Benford, D. J.; Staguhn, J. G.; Irwin, K. D.; Maddalena, R. J.; McMullin, J. P.; Shepherd, D.S.; Sievers, A.

    2009-11-01

    We have used the new 90 GHz MUSTANG camera on the Robert C. Byrd Green Bank Telescope (GBT) to map the bright Huygens region of the star-forming region M42 with a resolution of 9'' and a sensitivity of 2.8 mJy beam{sup -1}. Ninety GHz is an interesting transition frequency, as MUSTANG detects both the free-free emission characteristic of the H II region created by the Trapezium stars, normally seen at lower frequencies, and thermal dust emission from the background OMC1 molecular cloud, normally mapped at higher frequencies. We also present similar data from the 150 GHz GISMO camera taken on the IRAM 30 m telescope. This map has 15'' resolution. By combining the MUSTANG data with 1.4, 8, and 21 GHz radio data from the VLA and GBT, we derive a new estimate of the emission measure averaged electron temperature of T{sub e} = 11376 +- 1050 K by an original method relating free-free emission intensities at optically thin and optically thick frequencies. Combining Infrared Space Observatory-long wavelength spectrometer (ISO-LWS) data with our data, we derive a new estimate of the dust temperature and spectral emissivity index within the 80'' ISO-LWS beam toward Orion KL/BN, T{sub d} = 42 +- 3 K and beta {sub d} = 1.3 +- 0.1. We show that both T{sub d} and beta {sub d} decrease when going from the H II region and excited OMC1 interface to the denser UV shielded part of OMC1 (Orion KL/BN, Orion S). With a model consisting of only free-free and thermal dust emission, we are able to fit data taken at frequencies from 1.5 GHz to 854 GHz (350 mum).

  16. High blood pressure and visual sensitivity

    NASA Astrophysics Data System (ADS)

    Eisner, Alvin; Samples, John R.

    2003-09-01

    The study had two main purposes: (1) to determine whether the foveal visual sensitivities of people treated for high blood pressure (vascular hypertension) differ from the sensitivities of people who have not been diagnosed with high blood pressure and (2) to understand how visual adaptation is related to standard measures of systemic cardiovascular function. Two groups of middle-aged subjects-hypertensive and normotensive-were examined with a series of test/background stimulus combinations. All subjects met rigorous inclusion criteria for excellent ocular health. Although the visual sensitivities of the two subject groups overlapped extensively, the age-related rate of sensitivity loss was, for some measures, greater for the hypertensive subjects, possibly because of adaptation differences between the two groups. Overall, the degree of steady-state sensitivity loss resulting from an increase of background illuminance (for 580-nm backgrounds) was slightly less for the hypertensive subjects. Among normotensive subjects, the ability of a bright (3.8-log-td), long-wavelength (640-nm) adapting background to selectively suppress the flicker response of long-wavelength-sensitive (LWS) cones was related inversely to the ratio of mean arterial blood pressure to heart rate. The degree of selective suppression was also related to heart rate alone, and there was evidence that short-term changes of cardiovascular response were important. The results suggest that (1) vascular hypertension, or possibly its treatment, subtly affects visual function even in the absence of eye disease and (2) changes in blood flow affect retinal light-adaptation processes involved in the selective suppression of the flicker response from LWS cones caused by bright, long-wavelength backgrounds.

  17. ISO Key Project: Exploring the Full Range of Quasar/Agn Properties

    NASA Technical Reports Server (NTRS)

    Wilkes, Belinda; Oliversen, Ronald J. (Technical Monitor)

    2003-01-01

    While most of the work on this program has been completed, as previously reported, the portion of the program dealing with the subtopic of ISO LWS data analysis and reduction for the LWS Extragalactic Science Team and its leader, Dr. Howard Smith, is still active. This program in fact continues to generate results, and newly available computer modeling has extended the value of the datasets. As a result the team requests a one-year no-cost extension to this program, through 31 December 2004. The essence of the proposal is to perform ISO spectroscopic studies, including data analysis and modeling, of star-formation regions using an ensemble of archival space-based data from the Infrared Space Observatory's Long Wavelength Spectrometer and Short Wavelength Spectrometer, but including as well some other spectroscopic databases. Four kinds of regions are considered in the studies: (1) disks around more evolved objects; (2) young, low or high mass pre-main sequence stars in star-formation regions; (3) star formation in external, bright IR galaxies; and (4) the galactic center. One prime focus of the program is the OH lines in the far infrared. The program has the following goals: 1) Refine the data analysis of ISO observations to obtain deeper and better SNR results on selected sources. The ISO data itself underwent 'pipeline 10' reductions in early 2001, and additional 'hands-on data reduction packages' were supplied by the ISO teams in 2001. The Fabry-Perot database is particularly sensitive to noise and slight calibration errors; 2) Model the atomic and molecular line shapes, in particular the OH lines, using revised Monte-Carlo techniques developed by the SWAS team at the Center for Astrophysics; 3) Attend scientific meetings and workshops; 4) Perform E&PO activities related to infrared astrophysics and/or spectroscopy.

  18. ISO Key Project: Exploring The Full Range of Quasar/AGN Properties

    NASA Technical Reports Server (NTRS)

    Wilkes, Belinda; West, Donald K. (Technical Monitor)

    2002-01-01

    While most of the work on this program has been completed, as previously reported, the portion of the program dealing with the sub topic of ISO LWS data analysis and reduction for the LWS Extragalactic Science Team and its leader, Dr. Howard Smith, is still active. This program in fact continues to generate results, and newly available computer modeling has extended the value of the datasets, As a result the team has requested and been granted an obtained a no-cost extension to this program, through December 31, 2003. The essence of the proposal is to perform ISO spectroscopic studies, including data analysis and modeling, of star formation regions using an ensemble of archival space-based data from the Infrared Space Observatory's Long Wavelength Spectrometer and Short Wavelength Spectrometer, but including as well some other spectroscopic data bases. Four kinds of regions are considered in the studies: (1) disks around more evolved objects; (2) young, low or high mass pre-main sequence stars in star formation regions; (3) star formation in external, bright IR galaxies; and (4) the galactic center. One prime focus of the program is the OH lines in the far infrared. The program has the following goals: (1) refine the data analysis of ISO observations, to obtain deeper and better SNR results on selected sources. The ISO data itself underwent "pipeline 10" reductions in early 2001, and additional "hands-on data reduction packages" were supplied by the ISO teams in 2001. The Fabry-Perot database in particularly sensitive to noise can slight calibration errors. (2) model the atomic and molecular line shapes, in particular the OH lines, using revised Monte-Carlo techniques developed by the SWAS team at the Center for Astrophysics; (3) attend scientific meetings and workshops; (4) do E&PO activities related to infrared astrophysics and/or spectroscopy.

  19. Population dynamics and growth of the bivalve Choromytilus meridionalis (Kr.) at different tidal levels

    NASA Astrophysics Data System (ADS)

    Griffiths, Roberta J.

    1981-01-01

    Settlement, growth and reproductive output of a population of Choromytilus meridionalis have been monitored at different shore levels at Bailey's Cottage, False Bay, South Africa. Settlement was irregular, occurring at 4- to 6-year intervals, and confined to the sublittoral and lower littoral of rocky areas. Spat settled on the existing mussel bed and adjacent clean rock surfaces. Continual migration of young mussels up the shore took place during the first 1 to 1·5 years of growth until an even distribution up to 0·5 m above L.W.S. was achieved. Juveniles displaced older individuals by moving between them and forcing them off the rocks so that the majority of the adult population were eliminated from the bed within the first year after spat settlement. Mortality in individual cohorts was largely caused by strong wave action and competition for space. The density of individuals within the mussel bed was closely related to mean shell length. Growth rates varied with habitat and declined markedly with increasing height above L.W.S. Sexual maturity was attained at approximately 20 mm and reproductive output rose from 5 kJ year -1 at this length to 80 kJ year -1 at 100 mm shell length. Since packing densities were much higher in smaller individuals the annual gamete output assessed on an area basis, remained fairly constant as the mussels grew, and averaged 1392 g m -2 year -1 dry weight (31 320 kJ m -2 year -1). Energy expended as gonad output exceeded that due to mortality by a factor of 10.

  20. Living with a Star: New Opportunities in Sun-Climate Research

    NASA Technical Reports Server (NTRS)

    2003-01-01

    Living With a Star is a NASA initiative employing the combination of dedicated spacecraft with targeted research and modeling efforts to improve what we know of solar effects of all kinds on the Earth and its surrounding space environment, with particular emphasis on those that have significant practical impacts on life and society. The highest priority among these concerns is the subject of this report: the potential effects of solar variability on regional and global climate, including the extent to which solar variability has contributed to the well-documented warming of the Earth in the last 100 years. Understanding how the climate system reacts to external forcing from the Sun will also greatly improve our knowledge of how climate will respond to other climate drivers, including those of anthropogenic origin. A parallel element of the LWS program addresses solar effects on space weather : the impulsive emissions of charged particles, short-wave electromagnetic radiation and magnetic disturbances in the upper atmosphere and near-Earth environment that also affect life and society. These include a wide variety of solar impacts on aeronautics, astronautics, electric power transmission, and national defense. Specific examples are (1) the impacts of potentially- damaging high energy radiation and atomic particles of solar origin on satellites and satellite operations, spacecraft electronics systems and components, electronic communications, electric power distribution grids, navigational and GPS systems, and high altitude aircraft; and (2) the threat of sporadic, high-energy solar radiation to astronauts and high altitude aircraft passengers and crews. Elements of the LWS program include an array of dedicated spacecraft in near- Earth and near-Sun orbits that will closely study and observe both the Sun itself and the impacts of its variations on the Earth's radiation belts and magnetosphere, the upper atmosphere, and ionosphere. These spacecraft, positioned to study and monitor changing conditions in the Sun-Earth neighborhood, will also serve as sentinels of solar storms and impulsive events.

  1. Solitary-wave emission fronts, spectral chirping, and coupling to beam acoustic modes in RPIC simulation of SRS backscatter.

    SciTech Connect

    DuBois, D. F.; Yin, L.; Daughton, W. S.; Bezzerides, B.; Dodd, E. S.; Vu, H. X.

    2004-01-01

    Detailed diagnostics of quasi-2D RPIC simulations of backward stimulated Raman scattering (BSRS), from single speckles under putative NIF conditions, reveal a complex spatio-temporal behavior. The scattered light consists of localized packets, tens of microns in width, traveling toward the laser at an appreciable fraction of the speed of light. Sub pico-second reflectivity pulses occur as these packets leave the system. The LW activity consists of a front traveling with the light packets with a wake of free LWs traveling in the laser direction. The parametric coupling occurs in the front where the scattered light and LW overlap and are strongest. As the light leaves the plasma the LW quickly decays, liberating its trapped electrons. The high frequency part of the |n{sub e}(k,{omega})|{sup 2} spectrum, where n{sub e} is the electron density fluctuation, consists of a narrow streak or straight line with a slope that is the velocity of the parametric front. The time dependence of |n{sub e}(k,t)|{sup 2}, shows that during each pulse the most intense value of k also 'chirps' to higher values, consistent with the k excursions seen in the |n{sub e}(k,{omega})|{sup 2} spectrum. But k does not always return, in the subsequent pulses, to the original parametrically matched value, indicating that, in spite of side loss, the electron distribution function does not return to its original Maxwellian form. Liberated pulses of hot electrons result in down-stream, bump on tail distributions that excite LWs and beam acoustic modes deeper in the plasma. The frequency broadened spectra are consistent with Thomson scatter spectra observed in TRIDENT single-hot-spot experiments in the high k{lambda}{sub D}, trapping regime. Further details including a comparison of results from full PIC simulations, and movies of the spatio-temporal behavior, will be given in the poster by L Yin et al.

  2. 90 GHz and 150 GHz Observations of the Orion M42 Region. A Submillimeter to Radio Analysis

    NASA Technical Reports Server (NTRS)

    Dicker, S. R.; Mason, B. S.; Korngut, P. M.; Cotton, W. D.; Compiegne, M.; Devlin, M. J.; Martin, P. G.; Ade, P. A. R; Benford, D. J.; Irwin, K. D.; Maddalena, R. J.; McMullin, J. P.; Shepherd, D. S.; Sievers, A.; Staguhn, J. G.; Tucker, C.

    2009-01-01

    We have used the new 90GHz MUSTANG camera on the Robert C. Green Bank Telescope (GBT)to map the bright Huygens region of the star-forming region M42 with a resolution of 9" and a sensitivity of 2.8 mJy/beam. Ninety GHz is an interesting transition frequency, as MUSTANG detects both the free-free emission characteristic of the H II region created by the Trapezium stars, normally seen at lower frequencies, and thermal dust emission from the background OMCI molecular cloud, normally mapped at higher frequencies. We also present similar data from the 150 GHz GISMO camera taken on the IRAM 30 m telescope. This map has 15" resolution. By combining the MUSTANG data with 1.4, 8. and 31 GHz radio data from the VLA and GBT, we derive a new estimate of the emission measure averaged electron temperature of T(sub e) = 11376+/-1050 K by an original method relating free-free emission intensities at optically thin and optically thick frequencies. Combining Infrared Space Observatory-long wavelength spectrometer (ISO-LWS) data with our data, we derive a new estimate of the dust temperature and spectral emissivity index within the 80" ISO-LWS beam toward Orion KL/BN, T(sub d) = 42+/-3 K and Beta(sub d) = 1.3+/-0.1. We show that both T(sub d) and Beta(sub d) decrease when going from the H II region and excited OMCI interface to the denser UV shielded part OMCI (Orion KL/BN, Orion S). With a model consisting of only free-free and thermal dust emission, we are able to fit data taken at frequencies from 1.5 GHz to 854 GHz (350 micrometers).

  3. Spatiotemporal chaos and the dynamics of coupled Langmuir and ion-acoustic waves in plasmas.

    PubMed

    Banerjee, S; Misra, A P; Shukla, P K; Rondoni, L

    2010-04-01

    A simulation study is performed to investigate the dynamics of coupled Langmuir waves (LWs) and ion-acoustic waves (IAWs) in an unmagnetized plasma. The effects of dispersion due to charge separation and the density nonlinearity associated with the IAWs are considered to modify the properties of Langmuir solitons, as well as to model the dynamics of relatively large amplitude wave envelopes. It is found that the Langmuir wave electric field, indeed, increases by the effect of ion-wave nonlinearity (IWN). Use of a low-dimensional model, based on three Fourier modes, shows that a transition to temporal chaos is possible, when the length scale of the linearly excited modes is larger than that of the most unstable ones. The chaotic behaviors of the unstable modes are identified by the analysis of Lyapunov exponent spectra. The space-time evolution of the coupled LWs and IAWs shows that the IWN can cause the excitation of many unstable harmonic modes and can lead to strong IAW emission. This occurs when the initial wave field is relatively large or the length scale of IAWs is larger than the soliton characteristic size. Numerical simulation also reveals that many solitary patterns can be excited and generated through the modulational instability of unstable harmonic modes. As time goes on, these solitons are seen to appear in the spatially partial coherence state due to the free ion-acoustic radiation as well as in the state of spatiotemporal chaos due to collision and fusion in the stochastic motion. The latter results in the redistribution of initial wave energy into a few modes with small length scales, which may lead to the onset of Langmuir turbulence in laboratory as well as space plasmas. PMID:20481845

  4. Wetland treatment of oil and gas well waste waters. Final report

    SciTech Connect

    Kadlec, R.; Srinivasan, K.

    1995-08-01

    Constructed wetlands are small on-site systems that possess three of the most desirable components of an industrial waste water treatment scheme: low cost, low maintenance and upset resistance. The main objective of the present study is to extend the knowledge base of wetland treatment systems to include processes and substances of particular importance to small, on-site systems receiving oil and gas well wastewaters. A list of the most relevant and comprehensive publications on the design of wetlands for water quality improvement was compiled and critically reviewed. Based on our literature search and conversations with researchers in the private sector, toxic organics such as Phenolics and b-naphthoic acid, (NA), and metals such as CU(II) and CR(VI) were selected as target adsorbates. A total of 90 lysimeters equivalent to a laboratory-scale wetland were designed and built to monitor the uptake and transformation of toxic organics and the immobilization of metal ions. Studies on the uptake of toxic organics such as phenol and b-naphthoic acid (NA) and heavy metals such as Cu(II) and Cr(VI), the latter two singly or as non-stoichiometric mixtures by laboratory-type wetlands (LWs) were conducted. These LWs were designed and built during the first year of this study. A road map and guidelines for a field-scale implementation of a wetland system for the treatment of oil and gas wastewaters have been suggested. Two types of wetlands, surface flow (SF) and sub surface flow (SSF), have been considered, and the relative merits of each configuration have been reviewed.

  5. The pineal complex of the European sea bass (Dicentrarchus labrax): I. histological, immunohistochemical and qPCR study.

    PubMed

    Herrera-Pérez, P; Servili, A; Rendón, M C; Sánchez-Vázquez, F J; Falcón, J; Muñoz-Cueto, J A

    2011-04-01

    The pineal organ of fish is a photosensory and neuroendocrine epithalamic structure that plays a key role in the temporal organisation of physiological and behavioural processes. In this study performed in the European sea bass, Dicentrarchus labrax, we provided an in-depth description of the macroscopic and microscopic anatomy of the pineal organ and identified the presence of photoreceptor and presumed melatonin-producing cells using histological and immunohistochemical techniques. In addition, we analysed in the pineal the day-night expression (using quantitative real-time PCR) of two key enzymes in the melatonin-synthesising pathway; arylalkylamine-N-acetyltransferase 2 (AANAT2) and hydroxyindole-O-methyltransferase (HIOMT). The pineal complex of sea bass consisted of a narrow and short pineal stalk that adopts a vertical disposition, a small-sized pineal end vesicle firmly attached to the skull by connective tissue, a parapineal organ and a convoluted dorsal sac. Immunohistochemical study showed the presence of abundant serotonin-positive cells. Cone opsin-like and rod opsin-like photoreceptor cells were also evidenced in the pineal stalk and vesicle. Both Aanat2 and Hiomt were expressed in sea bass pineal organ. Aanat2 exhibited higher nocturnal transcript levels, while no significant day-night differences were found for Hiomt. These results, together with ongoing studies analysing neural and neurohormonal outputs from the pineal organ of sea bass, provide the basic framework to understand the transduction integration of light stimulus in this relevant species for marine aquaculture. PMID:21310229

  6. Absence of functional short-wavelength sensitive cone pigments in hamsters (Mesocricetus).

    PubMed

    Williams, Gary A; Jacobs, Gerald H

    2008-05-01

    Studies of Syrian golden hamsters (Mesocricetus auratus) have yielded contradictory evidence as to whether the retina of this species supports a population of cones containing short-wavelength sensitive pigments. We undertook a re-examination of this issue by (a) measuring lens transmission, (b) determining complete spectral sensitivity functions using electroretinogram (ERG) flicker photometry, (c) employing a sensitive chromatic-adaptation paradigm in conjunction with ERG measurements to conduct a specific search for the presence of a short-wavelength sensitive mechanism, and (d) assaying for the presence of retinal mRNA using real-time, reverse transcription polymerase chain reactions (RT-PCR). Parallel measurements were made on Turkish hamster (Mesocricetus brandtii) and control measurements were derived from recordings made on a rodent whose retina is known to contain a population of short-wavelength sensitive cones (the rat, Rattus norvegicus). Although UV opsin transcripts can be detected in the retina of the Syrian hamster, the electrophysiological measurements imply that these are not translated. Syrian hamsters thus lack a functional short-wavelength sensitive pigment, and that seems also true for the Turkish hamster. Members of this genus belong to a disparate group of mammals that have lost function of their short-wavelength sensitive cone pigments through ancestral opsin gene mutations. PMID:18259758

  7. Cone Health and Retinoids.

    PubMed

    Kono, Masahiro

    2015-01-01

    Cones are photoreceptor cells used for bright light and color vision. Retinoids are vitamin A derivatives, one of which is the 11-cis aldehyde form that serves as the chromophore for both cone and rod visual pigments. In the visual disease, Type 2 Leber congenital amaurosis (LCA2), 11-cis-retinal generation is inhibited or abolished. Work by others has shown that patients with LCA2 have symptoms consistent with degenerating cones. In mouse models for LCA2, early cone degeneration is readily apparent: cone opsins and other proteins associated with the outer segment are delocalized and cell numbers decline rapidly within the first month. Rods would appear normal morphologically and functionally, if not for the absence of chromophore. Supplementation of mouse models of LCA2 with cis-retinoids has been shown to slow loss of cone photoreceptor cells if mice were maintained in darkness. Thus, 11-cis-retinal appears not only to have a role in the light response reaction but also to promote proper trafficking of the cone opsins and maintain viable cones. PMID:26310171

  8. Isolation and characterization of melanopsin and pinopsin expression within photoreceptive sites of reptiles

    NASA Astrophysics Data System (ADS)

    Frigato, Elena; Vallone, Daniela; Bertolucci, Cristiano; Foulkes, Nicholas S.

    2006-08-01

    Non-mammalian vertebrates have multiple extraocular photoreceptors, mainly localised in the pineal complex and the brain, to mediate irradiance detection. In this study, we report the full-length cDNA cloning of ruin lizard melanopsin and pinopsin. The high level of identity with opsins in both the transmembrane regions, where the chromophore binding site is located, and the intracellular loops, where the G-proteins interact, suggests that both melanopsin and pinopsin should be able to generate a stable photopigment, capable of triggering a transduction cascade mediated by G-proteins. Phylogenetic analysis showed that both opsins are located on the expected branches of the corresponding sequences of ortholog proteins. Subsequently, using RT-PCR and RPA analysis, we verified the expression of ruin lizard melanopsin and pinopsin in directly photosensitive organs, such as the lateral eye, brain, pineal gland and parietal eye. Melanopsin expression was detected in the lateral eye and all major regions of the brain. However, different from the situation in Xenopus and chicken, melanopsin is not expressed in the ruin lizard pineal. Pinopsin mRNA expression was only detected in the pineal complex. As a result of their phylogenetic position and ecology, reptiles provide the circadian field with some of the most interesting models for understanding the evolution of the vertebrate circadian timing system and its response to light. This characterization of melanopsin and pinopsin expression in the ruin lizard will be important for future studies aimed at understanding the molecular basis of circadian light detection in reptiles.

  9. Self-organized criticality and color vision: A guide to water-protein landscape evolution

    NASA Astrophysics Data System (ADS)

    Phillips, J. C.

    2013-02-01

    We focus here on the scaling properties of small interspecies differences between red cone opsin transmembrane proteins, using a hydropathic elastic roughening tool previously applied to the rhodopsin rod transmembrane proteins. This tool is based on a non-Euclidean hydropathic metric realistically rooted in the atomic coordinates of 5526 protein segments, which thereby encapsulates universal non-Euclidean long-range differential geometrical features of water films enveloping globular proteins in the Protein Data Bank. Whereas the rhodopsin blue rod water films are smoothest in humans, the red cone opsins’ water films are optimized for smoothness in cats and elephants, consistent with protein species landscapes that evolve differently in different contexts. We also analyze red cone opsins in the chromatophore-containing family of chameleons, snakes, zebrafish and goldfish, where short- and long-range (BLAST and hydropathic) amino acid (aa) correlations are found with values as large as 97%-99%. We use hydropathic aa optimization to estimate the maximum number Nmax of color shades that the human eye can discriminate, and obtain 106

  10. Evidence for Multiple Phototransduction Pathways in a Reef-Building Coral

    PubMed Central

    Mason, Benjamin; Schmale, Michael; Gibbs, Patrick; Miller, Margaret W.; Wang, Qiang; Levay, Konstantin; Shestopalov, Valery; Slepak, Vladlen Z.

    2012-01-01

    Photosensitive behaviors and circadian rhythms are well documented in reef-building corals and their larvae, but the mechanisms responsible for photoreception have not been described in these organisms. Here we report the cloning, immunolocalization, and partial biochemical characterization of three opsins and four G proteins expressed in planulae of the Caribbean elkhorn coral, Acropora palmata. All three opsins (acropsins 1–3) possess conserved seven-pass transmembrane structure, and localize to distinct regions of coral planulae. Acropsin 1 was localized in the larval endoderm, while acropsin 2 was localized in solitary cells of the ectoderm. These rod-like cells displayed a remarkably polarized distribution, concentrated in the aboral end. We also cloned four A. palmata G protein alpha subunits. Three were homologs of vertebrate Gi, Go, and Gq. The fourth is presumably a novel G protein, which displays only 40% identity with the nearest known G protein, and we termed it Gc for “cnidarian”. We show that Gc and Gq can be activated by acropsins in a light-dependent manner in vitro. This indicates that at least acropsins 1 and 3 can form functional photoreceptors and potentially may play a role in color preference during settlement, vertical positioning and other light-guided behaviors observed in coral larvae. PMID:23227169

  11. The optogenetic (r)evolution.

    PubMed

    Rein, Martin L; Deussing, Jan M

    2012-02-01

    Optogenetics is a rapidly evolving field of technology that allows optical control of genetically targeted biological systems at high temporal and spatial resolution. By heterologous expression of light-sensitive microbial membrane proteins, opsins, cell type-specific depolarization or silencing can be optically induced on a millisecond time scale. What started in a petri dish is applicable today to more complex systems, ranging from the dissection of brain circuitries in vitro to behavioral analyses in freely moving animals. Persistent technical improvement has focused on the identification of new opsins, suitable for optogenetic purposes and genetic engineering of existing ones. Optical stimulation can be combined with various readouts defined by the desired resolution of the experimental setup. Although recent developments in optogenetics have largely focused on neuroscience it has lately been extended to other targets, including stem cell research and regenerative medicine. Further development of optogenetic approaches will not only highly increase our insight into health and disease states but might also pave the way for a future use in therapeutic applications. PMID:22183142

  12. The molecular genetics of red and green color vision in mammals.

    PubMed

    Yokoyama, S; Radlwimmer, F B

    1999-10-01

    To elucidate the molecular mechanisms of red-green color vision in mammals, we have cloned and sequenced the red and green opsin cDNAs of cat (Felis catus), horse (Equus caballus), gray squirrel (Sciurus carolinensis), white-tailed deer (Odocoileus virginianus), and guinea pig (Cavia porcellus). These opsins were expressed in COS1 cells and reconstituted with 11-cis-retinal. The purified visual pigments of the cat, horse, squirrel, deer, and guinea pig have lambdamax values at 553, 545, 532, 531, and 516 nm, respectively, which are precise to within +/-1 nm. We also regenerated the "true" red pigment of goldfish (Carassius auratus), which has a lambdamax value at 559 +/- 4 nm. Multiple linear regression analyses show that S180A, H197Y, Y277F, T285A, and A308S shift the lambdamax values of the red and green pigments in mammals toward blue by 7, 28, 7, 15, and 16 nm, respectively, and the reverse amino acid changes toward red by the same extents. The additive effects of these amino acid changes fully explain the red-green color vision in a wide range of mammalian species, goldfish, American chameleon (Anolis carolinensis), and pigeon (Columba livia). PMID:10511567

  13. Structure-function studies on bacteriorhodopsin. IX. Substitutions of tryptophan residues affect protein-retinal interactions in bacteriorhodopsin

    SciTech Connect

    Mogi, T.; Marti, T.; Khorana, H.G. )

    1989-08-25

    Bacteriorhodopsin contains 8 tryptophan residues distributed across the membrane-embedded helices. To study their possible functions, we have replaced them one at a time by phenylalanine; in addition, Trp-137 and -138 have been replaced by cysteine. The mutants were prepared by cassette mutagenesis of the synthetic bacterio-opsin gene, expression and purification of the mutant apoproteins, renaturation, and chromophore regeneration. The replacement of Trp-10, Trp-12 (helix A), Trp-80 (helix C), and Trp-138 (helix E) by phenylalanine and of Trp-137 and Trp-138 by cysteine did not significantly alter the absorption spectra or affect their proton pumping. However, substitution of the remaining tryptophans by phenylalanine had the following effects. (1) Substitution of Trp-86 (helix C) and Trp-137 gave chromophores blue-shifted by 20 nm and resulted in reduced proton pumping to about 30%. (2) As also reported previously, substitution of Trp-182 and Trp-189 (helix F) caused large blue shifts (70 and 40 nm, respectively) in the chromophore and affected proton pumping. (3) The substitution of Trp-86 and Trp-182 by phenylalanine conferred acid instability on these mutants. The spectral shifts indicate that Trp-86, Trp-182, Trp-189, and possibly Trp-137 interact with retinal. It is proposed that these tryptophans, probably along with Tyr-57 (helix B) and Tyr-185 (helix F), form a retinal binding pocket. We discuss the role of tryptophan residues that are conserved in bacteriorhodopsin, halorhodopsin, and the related family of opsin proteins.

  14. Preservation of cone photoreceptors after a rapid yet transient degeneration and remodeling in cone-only Nrl−/− mouse retina

    PubMed Central

    Roger, Jerome E; Ranganath, Keerthi; Zhao, Lian; Cojocaru, Radu I; Brooks, Matthew; Gotoh, Norimoto; Veleri, Shobi; Hiriyanna, Avinash; Rachel, Rivka A; Campos, Maria Mercedes; Fariss, Robert N; Wong, Wai T; Swaroop, Anand

    2012-01-01

    Cone photoreceptors are the primary initiator of visual transduction in the human retina. Dysfunction or death of rod photoreceptors precedes cone loss in many retinal and macular degenerative diseases, suggesting a rod-dependent trophic support for cone survival. Rod differentiation and homeostasis are dependent on the basic motif leucine zipper transcription factor NRL. The loss of Nrl (Nrl−/−) in mice results in a retina with predominantly S-opsin containing cones that exhibit molecular and functional characteristics of WT cones. Here we report that Nrl−/− retina undergoes a rapid but transient period of degeneration in early adulthood, with cone apoptosis, retinal detachment, alterations in retinal vessel structure, and activation and translocation of retinal microglia. However, cone degeneration stabilizes by four months of age, resulting in a thinner but intact outer nuclear layer with residual cones expressing S- and M-opsins and a preserved photopic ERG. At this stage, microglia translocate back to the inner retina and reacquire a quiescent morphology. Gene profiling analysis during the period of transient degeneration reveals misregulation of genes related to stress response and inflammation, implying their involvement in cone death. The Nrl−/− mouse illustrates the long-term viability of cones in the absence of rods and RPE defects in a rodless retina. We propose that Nrl−/− retina may serve as a model for elucidating mechanisms of cone homeostasis and degeneration that would be relevant to understanding diseases of the cone-dominant human macula. PMID:22238088

  15. Light-induced currents in Xenopus oocytes expressing bovine rhodopsin.

    PubMed Central

    Knox, B E; Khorana, H G; Nasi, E

    1993-01-01

    1. We have investigated the functioning of bovine rod opsin, which is efficiently synthesized from RNA made by in vitro transcription, following injection into Xenopus oocytes. We found that oocytes expressing the gene for opsin exhibit light-dependent ionic currents only after pigment generation by incubation with 11-cis-retinal. These currents are similar to the endogenous muscarinic acetylcholine (ACh) response of oocytes, but their amplitude is substantially smaller. 2. In order to optimize the conditions for obtaining light-induced currents in RNA-injected oocytes, the native ACh response was examined under several conditions. It was found that elevated external calcium markedly enhances the muscarinic response and that these currents have a non-linear dependence on membrane voltage, increasing substantially with depolarization. 3. Using the optimal conditions for evoking the largest ACh responses, (28 mM [Ca2+]o, 0 mV, omission of serum and Hepes from the media), the light-evoked currents obtained in RNA-injected oocytes were remarkably enhanced, and responses to multiple light stimuli could be obtained. 4. The light response appeared to desensitize, even after long periods of recovery and pigment regeneration. By contrast, the ACh responses continued to appear normal. These results suggest that desensitization of photoresponses expressed in Xenopus oocytes involve changes at early stages of the pathway, resulting in a reduced ability of rhodopsin to couple to the endogenous signalling system. Images Fig. 3 PMID:7692039

  16. Exorhodopsin and melanopsin systems in the pineal complex and brain at early developmental stages of Atlantic halibut (Hippoglossus hippoglossus).

    PubMed

    Eilertsen, Mariann; Drivenes, Oyvind; Edvardsen, Rolf B; Bradley, Clarrisa A; Ebbesson, Lars O E; Helvik, Jon Vidar

    2014-12-15

    The complexity of the nonvisual photoreception systems in teleosts has just started to be appreciated, with colocalization of multiple photoreceptor types with unresolved functions. Here we describe an intricate expression pattern of melanopsins in early life stages of the marine flat fish Atlantic halibut (Hippoglossus hippoglossus), a period when the unpigmented brain is directly exposed to environmental photons. We show a refined and extensive expression of melanopsins in the halibut brain already at the time of hatching, long before the eyes are functional. We detect melanopsin in the habenula, suprachiasmatic nucleus, dorsal thalamus, and lateral tubular nucleus of first feeding larvae, suggesting conserved functions of the melanopsins in marine teleosts. The complex expression of melanopsins already at larval stages indicates the importance of nonvisual photoreception early in development. Most strikingly, we detect expression of both exorhodopsin and melanopsin in the pineal complex of halibut larvae. Double-fluorescence labeling showed that two clusters of melanopsin-positive cells are located lateral to the central rosette of exorhodopsin-positive cells. The localization of different photopigments in the pineal complex suggests that two parallel photoreceptor systems may be active. Furthermore, the dispersed melanopsin-positive cells in the spinal cord of halibut larvae at the time of hatching may be primary sensory cells or interneurons representing the first example of dispersed high-order photoreceptor cells. The appearance of nonvisual opsins early in the development of halibut provides an alternative model for studying the evolution and functional significance of nonvisual opsins. PMID:25044160

  17. Development of transgenic animals for optogenetic manipulation of mammalian nervous system function: Progress and prospects for behavioral neuroscience

    PubMed Central

    Ting, Jonathan T.; Feng, Guoping

    2014-01-01

    Here we review the rapidly growing toolbox of transgenic mice and rats that exhibit functional expression of engineered opsins for neuronal activation and silencing with light. Collectively, these transgenic animals are enabling neuroscientists to access and manipulate the many diverse cell types in the mammalian nervous system in order to probe synaptic and circuitry connectivity, function, and dysfunction. The availability of transgenic lines affords important advantages such as stable and heritable transgene expression patterns across experimental cohorts. As such, the use of transgenic lines precludes the need for other costly and labor-intensive procedures to achieve functional transgene expression in each individual experimental animal. This represents an important consideration when large cohorts of experimental animals are desirable as in many common behavioral assays. We describe the diverse strategies that have been implemented for developing transgenic mouse and rat lines and highlight recent advances that have led to dramatic improvements in achieving functional transgene expression of engineered opsins. Furthermore, we discuss considerations and caveats associated with implementing recently developed transgenic lines for optogenetics-based experimentation. Lastly, we propose strategies that can be implemented to develop and refine the next generation of genetically modified animals for behaviorally-focused optogenetics-based applications. PMID:23473879

  18. Formation and Decay of the Arrestin·Rhodopsin Complex in Native Disc Membranes*

    PubMed Central

    Beyrière, Florent; Sommer, Martha E.; Szczepek, Michal; Bartl, Franz J.; Hofmann, Klaus Peter; Heck, Martin; Ritter, Eglof

    2015-01-01

    In the G protein-coupled receptor rhodopsin, light-induced cis/trans isomerization of the retinal ligand triggers a series of distinct receptor states culminating in the active Metarhodopsin II (Meta II) state, which binds and activates the G protein transducin (Gt). Long before Meta II decays into the aporeceptor opsin and free all-trans-retinal, its signaling is quenched by receptor phosphorylation and binding of the protein arrestin-1, which blocks further access of Gt to Meta II. Although recent crystal structures of arrestin indicate how it might look in a precomplex with the phosphorylated receptor, the transition into the high affinity complex is not understood. Here we applied Fourier transform infrared spectroscopy to monitor the interaction of arrestin-1 and phosphorylated rhodopsin in native disc membranes. By isolating the unique infrared signature of arrestin binding, we directly observed the structural alterations in both reaction partners. In the high affinity complex, rhodopsin adopts a structure similar to Gt-bound Meta II. In arrestin, a modest loss of β-sheet structure indicates an increase in flexibility but is inconsistent with a large scale structural change. During Meta II decay, the arrestin-rhodopsin stoichiometry shifts from 1:1 to 1:2. Arrestin stabilizes half of the receptor population in a specific Meta II protein conformation, whereas the other half decays to inactive opsin. Altogether these results illustrate the distinct binding modes used by arrestin to interact with different functional forms of the receptor. PMID:25847250

  19. Light-dependent activation of G proteins by two isoforms of chicken melanopsins.

    PubMed

    Torii, Masaki; Kojima, Daisuke; Nishimura, Akiyuki; Itoh, Hiroshi; Fukada, Yoshitaka

    2015-11-01

    In the chicken pineal gland, light stimuli trigger signaling pathways mediated by two different subtypes, Gt and G11. These G proteins may be activated by any of the three major pineal opsins, pinopsin, OPN4-1 and OPN4-2, but biochemical evidence for the coupling has been missing except for functional coupling between pinopsin and Gt. Here we investigated the relative expression levels and the functional difference among the three pineal opsins. In the chicken pineal gland, the pinopsin mRNA level was significantly more abundant than the others, of which the OPN4-2 mRNA level was higher than that of OPN4-1. In G protein activation assays, Gt was strongly activated by pinopsin in a light-dependent manner, being consistent with previous studies, and weakly activated by OPN4-2. Unexpectedly, illuminated OPN4-2 more efficiently activated G protein(s) that was endogenously expressed in HEK293T cells in culture. On the other hand, Gq, the closest analogue of G11, was activated only by OPN4-1 although the activity was relatively weak under these conditions. These results suggest that OPN4-1 and OPN4-2 couple with Gq and Gt, respectively. Two melanopsins, OPN4-1 and OPN4-2, appear to have acquired mutually different functions through the evolution. PMID:26411960

  20. Analysis of Conserved Glutamate and Aspartate Residues in Drosophila Rhodopsin 1 and Their Influence on Spectral Tuning.

    PubMed

    Zheng, Lijun; Farrell, David M; Fulton, Ruth M; Bagg, Eve E; Salcedo, Ernesto; Manino, Meridee; Britt, Steven G

    2015-09-01

    The molecular mechanisms that regulate invertebrate visual pigment absorption are poorly understood. Studies of amphioxus Go-opsin have demonstrated that Glu-181 functions as the counterion in this pigment. This finding has led to the proposal that Glu-181 may function as the counterion in other invertebrate visual pigments as well. Here we describe a series of mutagenesis experiments to test this hypothesis and to also test whether other conserved acidic amino acids in Drosophila Rhodopsin 1 (Rh1) may serve as the counterion of this visual pigment. Of the 5 Glu and Asp residues replaced by Gln or Asn in our experiments, none of the mutant pigments shift the absorption of Rh1 by more than 6 nm. In combination with prior studies, these results suggest that the counterion in Drosophila Rh1 may not be located at Glu-181 as in amphioxus, or at Glu-113 as in bovine rhodopsin. Conversely, the extremely low steady state levels of the E194Q mutant pigment (bovine opsin site Glu-181), and the rhabdomere degeneration observed in flies expressing this mutant demonstrate that a negatively charged residue at this position is essential for normal rhodopsin function in vivo. This work also raises the possibility that another residue or physiologic anion may compensate for the missing counterion in the E194Q mutant. PMID:26195627

  1. Constitutive phospholipid scramblase activity of a G protein-coupled receptor

    NASA Astrophysics Data System (ADS)

    Goren, Michael A.; Morizumi, Takefumi; Menon, Indu; Joseph, Jeremiah S.; Dittman, Jeremy S.; Cherezov, Vadim; Stevens, Raymond C.; Ernst, Oliver P.; Menon, Anant K.

    2014-10-01

    Opsin, the rhodopsin apoprotein, was recently shown to be an ATP-independent flippase (or scramblase) that equilibrates phospholipids across photoreceptor disc membranes in mammalian retina, a process required for disc homoeostasis. Here we show that scrambling is a constitutive activity of rhodopsin, distinct from its light-sensing function. Upon reconstitution into vesicles, discrete conformational states of the protein (rhodopsin, a metarhodopsin II-mimic, and two forms of opsin) facilitated rapid (>10,000 phospholipids per protein per second) scrambling of phospholipid probes. Our results indicate that the large conformational changes involved in converting rhodopsin to metarhodopsin II are not required for scrambling, and that the lipid translocation pathway either lies near the protein surface or involves membrane packing defects in the vicinity of the protein. In addition, we demonstrate that β2-adrenergic and adenosine A2A receptors scramble lipids, suggesting that rhodopsin-like G protein-coupled receptors may play an unexpected moonlighting role in re-modelling cell membranes.

  2. Computational modeling of cardiac optogenetics: Methodology overview & review of findings from simulations.

    PubMed

    Boyle, Patrick M; Karathanos, Thomas V; Entcheva, Emilia; Trayanova, Natalia A

    2015-10-01

    Cardiac optogenetics is emerging as an exciting new potential avenue to enable spatiotemporally precise control of excitable cells and tissue in the heart with low-energy optical stimuli. This approach involves the expression of exogenous light-sensitive proteins (opsins) in target heart tissue via viral gene or cell delivery. Preliminary experiments in optogenetically-modified cells, tissue, and organisms have made great strides towards demonstrating the feasibility of basic applications, including the use of light stimuli to pace or disrupt reentrant activity. However, it remains unknown whether techniques based on this intriguing technology could be scaled up and used in humans for novel clinical applications, such as pain-free optical defibrillation or dynamic modulation of action potential shape. A key step towards answering such questions is to explore potential optogenetics-based therapies using sophisticated computer simulation tools capable of realistically representing opsin delivery and light stimulation in biophysically detailed, patient-specific models of the human heart. This review provides (1) a detailed overview of the methodological developments necessary to represent optogenetics-based solutions in existing virtual heart platforms and (2) a survey of findings that have been derived from such simulations and a critical assessment of their significance with respect to the progress of the field. PMID:26002074

  3. Melanopsin and the Non-visual Photochemistry in the Inner Retina of Vertebrates.

    PubMed

    Díaz, Nicolás M; Morera, Luis P; Guido, Mario E

    2016-01-01

    Melanopsin (Opn4), a member of the G-protein-coupled receptor family, is a vitamin A-based opsin in the vertebrate retina that has been shown to be involved in the synchronization of circadian rhythms, pupillary light reflexes, melatonin suppression and other light-regulated tasks. In nonmammalian vertebrates there are two Opn4 genes, Opn4m and Opn4x, the mammalian and Xenopus orthologs respectively. Opn4x is only expressed in nonmammalian vertebrates including reptiles, fish and birds, while Opn4m is found in a subset of retinal ganglion cells (RGCs), the intrinsically photosensitive (ip) RGCs of the inner retina of both mammals and nonmammalian vertebrates. All opsins described utilize retinaldehyde as chromophore, photoisomerized from 11-cis- to all-trans-retinal upon light exposure. Visual retinal photoreceptor cones and rods, responsible for day and night vision respectively, recycle retinoids through a process called the visual cycle that involves the retinal pigment epithelium or glial Müller cells. Although Opn4 has been characterized as a bistable photopigment, little is known about the mechanism/s involved in its chromophore regeneration. In this review, we will attempt to shed light on the visual cycle taking place in the inner retina and discuss the state of the art in the nonvisual photochemistry of vertebrates. PMID:26500165

  4. Patterns of cell proliferation and rod photoreceptor differentiation in shark retinas.

    PubMed

    Ferreiro-Galve, Susana; Rodríguez-Moldes, Isabel; Anadón, Ramón; Candal, Eva

    2010-01-01

    We studied the pattern of cell proliferation and its relation with photoreceptor differentiation in the embryonic and postembryonic retina of two elasmobranchs, the lesser spotted dogfish (Scyliorhinus canicula) and the brown shyshark (Haploblepharus fuscus). Cell proliferation was studied with antibodies raised against proliferating cell nuclear antigen (PCNA) and phospho-histone-H3, and early photoreceptor differentiation with an antibody raised against rod opsin. As regards the spatiotemporal distribution of PCNA-immunoreactive cells, our results reveal a gradual loss of PCNA that coincides in a spatiotemporal sequence with the gradient of layer maturation. The presence of a peripheral growth zone containing pure-proliferating retinal progenitors (the ciliary marginal zone) in the adult retina matches with the general pattern observed in other groups of gnathostomous fishes. However, in the shark retina the generation of new cells is not restricted to the ciliary marginal zone but also occurs in retin