Opsin Transcripts Research Articles

Light-specific wavelengths differentially affect the exploration rate, opercular beat, skin color change, opsin transcripts, and the oxi-redox system of the longsnout seahorse Hippocampus reidi

Light is a strong stimulus for the sensory and endocrine systems. The opsins constitute a large family of proteins that can respond to specific light wavelengths. Hippocampus reidi is a near-threatened seahorse that has a diverse color pattern and sexual dimorphism. Over the years, H. reidi's unique characteristics, coupled with its high demand and over-exploitation for the aquarium trade, have raised concerns about its conservation, primarily due to their significant impact on wild populations. Here, we characterized chromatophore types in juvenile and adult H. reidi in captivity, and the effects of specific light wavelengths with the same irradiance (1.20 mW/cm2) on color change, growth, and survival rate. The xanthophores and melanophores were the major components of H. reidi pigmentation with differences in density and distribution between life stages and sexes. In the eye and skin of juveniles, the yellow (585 nm) wavelength induced a substantial increase in melanin levels compared to the individuals kept under white light (WL), blue (442 nm), or red (650 nm) wavelengths. In addition, blue and yellow wavelengths led to a higher juvenile mortality rate in comparison to the other treatments. Adult seahorses showed a rhythmic color change over 24 h, the highest reflectance values were obtained in the light phase, representing a daytime skin lightening for individuals under WL, blue and yellow wavelength, with changes in the acrophase. The yellow wavelength was more effective on juvenile seahorse pigmentation, while the blue wavelength exerted a stronger effect on the regulation of adult physiological color change. Dramatic changes in the opsin mRNA levels were life stage-dependent, which may infer ontogenetic opsin functions throughout seahorses' development. Exposure to specific wavelengths differentially affected the opsins mRNA levels in the skin and eyes of juveniles. In the juveniles, skin transcripts of visual (rh1, rh2, and lws) and non-visual opsins (opn3 and opn4x) were higher in individuals under yellow light. While in the juvenile's eyes, only rh1 and rh2 had increased transcripts influenced by yellow light; the lws and opn3 mRNA levels were higher in juveniles' eyes under WL. Prolonged exposure to yellow wavelength stimulates a robust increase in the antioxidant enzymes sod1 and sod2 mRNA levels. Our findings indicate that changes in the visible light spectrum alter physiological processes at different stages of life in H. reidi and may serve as the basis for a broader discussion about the implications of artificial light for aquatic species in captivity.

Increasing complexity of opsin expression across stomatopod development.

Stomatopods are well studied for their unique visual systems, which can consist of up to 16 different photoreceptor types and 33 opsin proteins expressed in the adults of some species. The light-sensing abilities of larval stomatopods are comparatively less well understood with limited information about the opsin repertoire of these early-life stages. Early work has suggested that larval stomatopods may not possess the extensive light detection abilities found in their adult counterparts. However, recent studies have shown that these larvae may have more complex photosensory systems than previously thought. To examine this idea at the molecular level, we characterized the expression of putative light-absorbing opsins across developmental stages, from embryo to adult, in the stomatopod species Pullosquilla thomassini using transcriptomic methods with a special focus on ecological and physiological transition periods. Opsin expression during the transition from the larval to the adult stage was further characterized in the species Gonodactylaceus falcatus. Opsin transcripts from short, middle, and long wavelength-sensitive clades were found in both species, and analysis of spectral tuning sites suggested differences in absorbance within these clades. This is the first study to document the changes in opsin repertoire across development in stomatopods, providing novel evidence for light detection across the visual spectrum in larvae.

Two mechanisms of retinal photoreceptor plasticity underlie rapid adaptation to novel light environments.

Fishes experience different light environments over short time periods that may require quick modulation of photoreceptor properties to optimize visual function. Previous research has shown that the relative expression of different visual pigment protein (opsin) transcripts can change within several days following exposure to new light environments, but whether such changes are mirrored by analogous modulation in opsin protein expression is unknown. Here, Atlantic halibut larvae and juveniles raised under white light were exposed to blue light for 1week and their retina compared to that of controls, which remained under white light. Blue light-treated larvae showed increased expression of all cone opsin transcripts, except rh2, over controls. They also had longer outer segments, and higher density of long wavelength sensitive (L) cones in the dorsal retina. In contrast, only the lws transcript was upregulated in juveniles exposed to blue light compared to controls but their L cone density was greater throughout the retina. These results demonstrate two mechanisms of rapid photoreceptor plasticity as a function of developmental stage associated with improved perception of achromatic or chromatic contrasts in line with the animal's ecological needs.

Evolutionary conservation of opsin gene expression patterns in the compound eyes of darkling beetles.

Recent large-scale studies of opsin gene contents in representatives of the largest order of insects, the Coleoptera (beetles), revealed that the blue wavelength-sensitive (B) opsin subfamily is absent in this clade, while the ultraviolet- (UV) and long wavelength-sensitive (LW) opsin subfamilies are broadly conserved with gene duplications possibly reintroducing blue sensitivity in select subclades. Little is known yet, however, how opsin genes are expressed in the compound eyes of beetles. In a previous study, we analyzed opsin gene expression in the red flour beetle Tribolium castaneum, a member of the family of darkling beetles (Tenebrionidae), and found that a singleton LW opsin homolog is homogeneously expressed in all photoreceptors of the compound eye retina with a singleton UV opsin homolog being co-expressed in the R7 subtype photoreceptors. To probe for the evolutionary conservation of these expression patterns, we isolated complete opsin transcript sequences from three additional species in the subfamily Tenebrionidae (Tribolium confusum, Tenebrio molitor, Zophobas morio) and studied their expression via whole mount in situ hybridization in the pupal retina. These experiments revealed very similar, if not identical, photoreceptor subtype-specific expression patterns in all three species compared with T. castaneum. Documenting a deep conservation of photoreceptor subtype-specific opsin gene expression in this range of darkling beetles, our study provides a first point of reference for broader comparative studies of retinal organization in the Coleoptera.

Analysis of Early Cone Dysfunction in an In Vivo Model of Rod-Cone Dystrophy.

Retinitis pigmentosa (RP) is a generic term for a group of genetic diseases characterized by loss of rod and cone photoreceptor cells. Although the genetic causes of RP frequently only affect the rod photoreceptor cells, cone photoreceptors become stressed in the absence of rods and undergo a secondary degeneration. Changes in the gene expression profile of cone photoreceptor cells are likely to occur prior to observable physiological changes. To this end, we sought to achieve greater understanding of the changes in cone photoreceptor cells early in the degeneration process of the Rho−/− mouse model. To account for gene expression changes attributed to loss of cone photoreceptor cells, we normalized PCR in the remaining number of cones to a cone cell reporter (OPN1-GFP). Gene expression profiles of key components involved in the cone phototransduction cascade were correlated with tests of retinal cone function prior to cell loss. A significant downregulation of the photoreceptor transcription factor Crx was observed, which preceded a significant downregulation in cone opsin transcripts that coincided with declining cone function. Our data add to the growing understanding of molecular changes that occur prior to cone dysfunction in a model of rod-cone dystrophy. It is of interest that gene supplementation of CRX by adeno-associated viral vector delivery prior to cone cell loss did not prevent cone photoreceptor degeneration in this mouse model.

Exceptional diversity of opsin expression patterns in Neogonodactylus oerstedii (Stomatopoda) retinas

Stomatopod crustaceans possess some of the most complex animal visual systems, including at least 16 spectrally distinct types of photoreceptive units (e.g., assemblages of photoreceptor cells). Here we fully characterize the set of opsin genes expressed in retinal tissues and determine expression patterns of each in the stomatopod Neogonodactylus oerstedii Using a combination of transcriptome and RACE sequencing, we identified 33 opsin transcripts expressed in each N. oerstedii eye, which are predicted to form 20 long-wavelength-sensitive, 10 middle-wavelength-sensitive, and three UV-sensitive visual pigments. Observed expression patterns of these 33 transcripts were highly unusual in five respects: 1) All long-wavelength and short/middle-wavelength photoreceptive units expressed multiple opsins, while UV photoreceptor cells expressed single opsins; 2) most of the long-wavelength photoreceptive units expressed at least one middle-wavelength-sensitive opsin transcript; 3) the photoreceptors involved in spatial, motion, and polarization vision expressed more transcripts than those involved in color vision; 4) there is a unique opsin transcript that is expressed in all eight of the photoreceptive units devoted to color vision; and 5) expression patterns in the peripheral hemispheres of the eyes suggest visual specializations not previously recognized in stomatopods. Elucidating the expression patterns of all opsin transcripts expressed in the N. oerstedii retina reveals the potential for previously undocumented functional diversity in the already complex stomatopod eye and is a first step toward understanding the functional significance of the unusual abundance of opsins found in many arthropod species' visual systems.

Straying from the flatfish retinal plan: Cone photoreceptor patterning in the common sole (Solea solea) and the Senegalese sole (Solea senegalensis).

The retinas of nonmammalian vertebrates have cone photoreceptor mosaics that are often organized as highly patterned lattice-like distributions. In fishes, the two main lattice-like patterns are composed of double cones and single cones that are either assembled as interdigitized squares or as alternating rows. The functional significance of such orderly patterning is unknown. Here, the cone mosaics in two species of Soleidae flatfishes, the common sole and the Senegalese sole, were characterized and compared to those from other fishes to explore variability in cone patterning and how it may relate to visual function. The cone mosaics of the common sole and the Senegalese sole consisted of single, double, and triple cones in formations that differed from the traditional square mosaic pattern reported for other flatfishes in that no evidence of higher order periodicity was present. Furthermore, mean regularity indices for single and double cones were conspicuously lower than those of other fishes with "typical" square and row mosaics, but comparable to those of goldfish, a species with lattice-like periodicity in its cone mosaic. Opsin transcripts detected by quantitative polymerase chain reaction (sws1, sws2, rh2.3, rh2.4, lws, and rh1) were uniformly expressed across the retina of the common sole but, in the Senegalese sole, sws2, rh2.4, and rh1 were more prevalent in the dorsal retina. Microspectrophotometry revealed five visual pigments in the retina of the common sole [S(472), M(523), M(536), L(559), and rod(511)] corresponding to the repertoire of transcripts quantified except for sws1. Overall, these results indicate a loss of cone mosaic patterning in species that are primarily nocturnal or dwell in low light environments as is the case for the common sole and the Senegalese sole. The corollary is that lattice-like patterning of the cone mosaic may improve visual acuity. Ecological and physiological correlates derived from observations across multiple fish taxa that live in low light environments and do not possess lattice-like cone mosaics are congruent with this claim.

The light at the end of the tunnel: photosensitivity in larvae of the mountain pine beetle (Coleoptera: Curculionidae: Scolytinae)

AbstractInvestigations of light sensitivity and its physiological effects on insects developing within subcortical tree tissues are limited, presumably due to the assumption that cryptic microhabitats are completely devoid of light. In this study, we documented light-mediated behaviour and opsin gene expression in larvae of the mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae: Scolytinae), an extremely important forest insect that is well adapted for development beneath the bark of pine (Pinus Linnaeus; Pinaceae) trees and is eyeless in the larval stage. Larvae were negatively phototactic, as they selected dark over light microhabitats in phototaxis assays. We recovered long-wavelength opsin transcripts from all life stages, including eggs and larvae, suggesting that D. ponderosae is photosensitive throughout its entire life cycle. Our results imply that photosensitivity contributes to immature D. ponderosae survival and that extraocular photoreception could be common among bark beetle larvae.

Isolation of photoreceptors from mature, developing, and regenerated zebrafish retinas, and of microglia/macrophages from regenerating zebrafish retinas

This paper describes experimental procedures for the dissociation of retinal cells of the zebrafish (Danio rerio) for subsequent fluorescence-activated cell sorting (FACS) and gene expression studies. Methods for dissociation of zebrafish retinas followed by FACS and RNA isolation were optimized. This methodology was applied to isolate pure sorted samples of rods, long wavelength-sensitive (LWS) cones, medium wavelength-sensitive (MWS; RH2-2) cones, short wavelength-sensitive (SWS2) cones, and UV-sensitive (SWS1) cones from retinas obtained at selective life-history stages of the zebrafish, and for some of these photoreceptors, following retinal regeneration. We also successfully separated lws1-expressing and lws2-expressing LWS cones from fish of a transgenic line in which lws1 is reported with green fluorescence protein (GFP) and lws2 is reported with red fluorescence protein (RFP). Microglia/macrophages were successfully sorted from regenerating retinas (7 days after a cytotoxic lesion) of a transgenic line in which these immune cells express GFP. Electropherograms verified downstream isolation of high-quality RNA from sorted samples. Examples of post-sorting analysis, as well as results of qRT-PCR studies, validated the purity of sorted populations. For example, qRT-PCR samples derived from isolated Rh2-2 cones contained detectable rh2-2 (opn1mw2) opsin transcripts, but lws opsin transcripts (lws1/opn1lw1, lws2/opn1lw2) were not detected, suggesting that the procedure likely separated double cone pairs. Through this method, pure, sorted cell samples can provide RNA that is reliable for downstream gene expression analyses, such as qRT-PCR and RNA-seq, which may reveal molecular signatures of photoreceptors and microglia for comparative transcriptomics studies.

Sequence, Structure, and Expression of Opsins in the Monochromatic Stomatopod Squilla empusa.

Most stomatopod crustaceans have complex retinas in their compound eyes, with up to 16 spectral types of photoreceptors, but members of the superfamily Squilloidea have much simpler retinas, thought to contain a single photoreceptor spectral class. In the Atlantic stomatopod Squilla empusa, microspectrophotometry shows that all photoreceptors absorb light maximally at 517 nm, indicating that a single visual pigment is present in all photoreceptors in the retina. However, six distinct, but partial, long wavelength sensitive (LWS) opsin transcripts, which encode the protein component of the visual pigment, have been previously isolated through RT-PCR. In order to investigate the spectral and functional differences among S. empusa's opsins, we used RT-PCR to complete the 3' end of sequences for five of the six expressed opsins. The extended sequences spanned from the first transmembrane (TM1) helix to the 3' end of the coding region. Using homology-based modeling, we predicted the three-dimensional structure of the amino acid translation of the S. empusa opsins. Based on these analyses, S. empusa LWS opsins share a high sequence identity in TM regions and in amino acids within 15 Å of the chromophore-binding lysine on TM helix 7 (TM7), suggesting that these opsins produce spectrally similar visual pigments in agreement with previous results. However, we propose that these spectrally similar opsins differ functionally, as there are non-conservative amino acid substitutions found in intracellular loop 2 (ICL2) and TM5/ICL3, which are critical regions for G-protein binding, and substitutions in extracellular regions suggest different chromophore attachment affinities. In situ hybridization of two of the opsins (Se5 and Se6) revealed strong co-expression in all photoreceptors in both midband and peripheral regions of the retina as well as in selected ocular and cerebral ganglion neuropils. These data suggest the expression of multiple opsins-likely spectrally identical, but functionally different-in multiple types of neuronal cells in S. empusa. This suggests that the multiple opsins characteristic of other stomatopod species may have similar functional specialization.

Variation in opsin transcript expression explains intraretinal differences in spectral sensitivity of the northern anchovy.

Vertebrate retinal photoreceptors house visual pigments that absorb light to begin the process of vision. The light absorbed by a visual pigment depends on its two molecular components: protein (opsin) and chromophore (a vitamin A derivative). Although an increasing number of studies show intraretinal variability in visual pigment content, it is only for two mammals (human and mouse) and two birds (chicken and pigeon) that such variability has been demonstrated to underlie differences in spectral sensitivity of the animal. Here, we show that the spectral sensitivity of the northern anchovy varies with retinal quadrant and that this variability can be explained by differences in the expression of opsin transcripts. Retinal (vitamin A1) was the only chromophore detected in the retina, ruling out this molecular component as a source of variation in spectral sensitivity. Chromatic adaptation experiments further showed that the dorsal retina had the capacity to mediate color vision. Together with published results for the ventral retina, this study is the first to demonstrate that intraretinal opsin variability in a fish drives corresponding variation in the animal's spectral sensitivity.

Fine-tuning light sensitivity in the starry flounder (Platichthys stellatus) retina: Regional variation in photoreceptor cell morphology and opsin gene expression.

Vertebrate color vision relies on the differential expression of visual pigment proteins (opsins) in cone photoreceptors of the retina. The diversity of opsins and their retinal expression patterns appear greatest for animals that experience variable light habitats, as is the case with flatfishes. Yet, opsin repertoires and expression patterns in this group of fishes are poorly described. Here, we unveil the visual opsin expression patterns of juvenile starry flounder (Platichthys stellatus) and describe the localization of cone types, their visual pigments and opsin expression. Juvenile starry flounder express eight opsins (Rh1, Sws1, Sws2A1, Sws2A2, Sws2B, Rh2A1, Rh2A2, Lws) and possess a corresponding number of photoreceptor visual pigments, with peak absorbance ranging from 369 to 557 nm. Retinal (vitamin A1) was the only chromophore detected in the retina. Intraretinal variation in opsin abundance consisted of greater expression of both RH2, and lesser expression of SWS1 and both SWS2A, opsin transcripts in the dorsal compared to the ventral retina. Overall cone density was greater in the dorsal retina which was also characterized by a larger proportion of unequal double cones compared with the ventral retina. Together, our results suggest that large opsin repertoires serve to optimize visual function under variable light environments by differential expression of opsin subsets with retinal location.

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

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.

Visual system evolution and the nature of the ancestral snake.

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.

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

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.

Seeing through the skin: dermal light sensitivity provides cryptism in moorish gecko

AbstractConcealment by means of colour change is a pre‐eminent deceptive mechanism used by both predators and prey. The moorish gecko Tarentola mauritanica is able to blend into the background by either darkening or paling according to the substrate darkness. Here we examined the functioning of background perception in moorish gecko. We experimentally excluded the involvement of melanophore‐stimulating hormone in camouflage. Blindfolded individuals change their colour consistently with the background. Surprisingly, individuals with covered flanks were not able to change colour, no matter whether they were allowed to see the substrate or not. Accordingly, we found high levels of opsin transcript and protein in the flank region of the gecko. These observations suggest that T. mauritanica skin melanophores are able to activate a process of colour change autonomously. This study yields the first evidence of crypsis mediated by dermal light sensitivity in amniotes.

The Evolution of Complexity in the Visual Systems of Stomatopods: Insights from Transcriptomics

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.

Evidence for a Phototransduction Cascade in an Early Brachiopod Embryo

Bilaterian photoreceptor cells are characterized by the expression of opsins, signal transduction genes, and ion channels, which together facilitate behavioral responses to light. We have previously identified a ciliary opsin gene from the brachiopod Terebratalia transversa, whose expression in gastrula stage embryos coincides with a photoresponse behavior, suggesting the presence of a functional phototransduction system in these early embryos. To further evaluate the potential for light reception in these embryos, we surveyed transcriptome data to identify phototransduction genes and evaluated their expression. In addition to the previously described ciliary opsin gene, we have identified two Go-class opsins that are also expressed in gastrula stage embryos. Representative members from all classes of Gα-protein genes were also expressed, with a Gα12-class gene being localized in the same anterior ectodermal domain as the opsin transcripts. Both CNG-class and TRP-class ion channels were expressed in the gastrula stage embryos, as were GRK and arrestin genes, which are associated with inhibition of rhodopsin activity. Taken together, these data support the presence of a functional phototransduction system in the early brachiopod embryo.

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

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. 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. 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. 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.

Complexity compounded: The evolution of phototransduction in stomatopods

Event Abstract Back to Event Complexity compounded: The evolution of phototransduction in stomatopods Megan L. Porter1*, Daniel I. Speiser2, Alexander K. Zaharoff2, Roy L. Caldwell3, Todd H. Oakley2 and Thomas W. Cronin4 1 University of University Baltimore County, Biological Sciences, United States 2 University of California Santa Barbara, Ecology, Evolution, and Marine Biology, United States 3 University of California, Berkeley, Integrative Biology, United States 4 University of Maryland Baltimore County, Biological Sciences, United States Stomatopod crustaceans have complex visual systems containing up to 16 different spectral classes of photoreceptors. Complexity is also found at the molecular level, with a previous study of stomatopod opsin genes describing many more expressed transcripts than predicted based on photoreceptor physiology. How other phototransduction cascade components contribute to stomatopod visual system complexity is still unknown. This study continues research on stomatopod visual systems by investigating additional components of the G-protein coupled receptor (GPCR) phototransduction cascade using retinal transcriptome datasets (generated using 454 or Illumina platforms) from four species: Hemisquilla californiensis, Neogonodactylus oerstedii, Pseudosquilla ciliata, and Squilla empusa. These species represent taxonomic and visual system diversity within the order Stomatopoda. Using Drosophila melanogaster genes as queries, assembled transcriptomes from each species were searched for phototransduction transcripts using reciprocal BLAST methods. Based on these searches, transcripts from genes specifying 13 different phototransduction components were identified from stomatopod retinal transcriptomes; however, not all 13 genes were identified in any single species. Identified phototransduction genes include visual arrestins, all subunits of the heterotrimeric G-protein (i.e. Gαq, Gβ, Gγ), phospholipase C, G protein-coupled receptor kinases 1 and 2, transient receptor potential (TRP) channels, and opsin transcripts. The set of expressed transduction genes suggest that stomatopods utilize a Gq mediated GPCR signaling cascade in their retinal photoreceptors. Two of the phototransduction genes – arrestin and opsin – were identified in the assemblies of all four species. In particular, the opsin contigs identified expand known stomatopod diversity to short-wavelength and ultraviolet sensitive arthropod clades, and repeat the results of previous studies that recovered more opsin transcripts than spectrally distinct types of photoreceptors. Keywords: phototransduction, Stomatopoda, Opsin, GPCR signal transduction, visual system evolution, retinal transcriptomics Conference: International Conference on Invertebrate Vision, Fjälkinge, Sweden, 1 Aug - 8 Aug, 2013. Presentation Type: Oral presentation preferred Topic: Transduction, signalling and coding Citation: Porter ML, Speiser DI, Zaharoff AK, Caldwell RL, Oakley TH and Cronin TW (2019). Complexity compounded: The evolution of phototransduction in stomatopods. Front. Physiol. Conference Abstract: International Conference on Invertebrate Vision. doi: 10.3389/conf.fphys.2013.25.00010 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 28 Feb 2013; Published Online: 09 Dec 2019. * Correspondence: Dr. Megan L Porter, University of University Baltimore County, Biological Sciences, Baltimore, MD, 21250, United States, Megan.Porter@usd.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Megan L Porter Daniel I Speiser Alexander K Zaharoff Roy L Caldwell Todd H Oakley Thomas W Cronin Google Megan L Porter Daniel I Speiser Alexander K Zaharoff Roy L Caldwell Todd H Oakley Thomas W Cronin Google Scholar Megan L Porter Daniel I Speiser Alexander K Zaharoff Roy L Caldwell Todd H Oakley Thomas W Cronin PubMed Megan L Porter Daniel I Speiser Alexander K Zaharoff Roy L Caldwell Todd H Oakley Thomas W Cronin Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.