Functional characterisation of the chromatically antagonistic photosensitive mechanism of erythrophores in the tilapia Oreochromis niloticus.

Non-visual photoreceptors with diverse photopigments allow organisms to adapt to changing light conditions. Whereas visual photoreceptors are involved in image formation, non-visual photoreceptors mainly undertake various non-image-forming tasks. They form specialised photosensory systems that measure the quality and quantity of light and enable appropriate behavioural and physiological responses. Chromatophores are dermal non-visual photoreceptors directly exposed to light and they not only receive ambient photic input but also respond to it. These specialised photosensitive pigment cells enable animals to adjust body coloration to fit environments, and play an important role in mate choice, camouflage and ultraviolet (UV) protection. However, the signalling pathway underlying chromatophore photoresponses and the physiological importance of chromatophore colour change remain under-investigated. Here, we characterised the intrinsic photosensitive system of red chromatophores (erythrophores) in tilapia. Like some non-visual photoreceptors, tilapia erythrophores showed wavelength-dependent photoresponses in two spectral regions: aggregations of inner pigment granules under UV and short-wavelengths and dispersions under middle- and long-wavelengths. The action spectra curve suggested that two primary photopigments exert opposite effects on these light-driven processes: SWS1 (short-wavelength sensitive 1) for aggregations and RH2b (rhodopsin-like) for dispersions. Both western blot and immunohistochemistry showed SWS1 expression in integumentary tissues and erythrophores. The membrane potential of erythrophores depolarised under UV illumination, suggesting that changes in membrane potential are required for photoresponses. These results suggest that SWS1 and RH2b play key roles in mediating intrinsic erythrophore photoresponses in different spectral ranges and this chromatically dependent antagonistic photosensitive mechanism may provide an advantage to detect subtle environmental photic change.

Retinal region of polarization sensitivity switches during ontogeny of rainbow trout.

Polarization sensitivity (PS) in vertebrate vision is controversial, perhaps because its underlying mechanism has remained obscure. An issue that might have added to the controversy is that rainbow trout (Oncorhynchus mykiss), which have served as the primary model system for polarization-based orientation, lose their ability to orient relative to celestial polarized-light patterns when parr (fry) transform into migratory smolts (juveniles), which would benefit most from polarization-based orientation. Here we addressed two key questions: (1) what is the mechanism underling PS?, and (2) how can the paradoxical loss of PS in trout smolts be reconciled? We assessed PS from optic nerve recordings in parr and smolts and found that the retinal region with enhanced PS shifted from the ventral retina in parr to the dorsal retina in smolts. This adaptation may allow fish to use the most reliable polarization field encountered at each life stage, the celestial polarization field in the shallow-swimming parr and the depth-insensitive underwater polarization field in the deep-swimming smolts. In addition, we assessed spectral sensitivity across the retina and during ontogeny and fit a cascade retinal model to PS data. We found that differential contribution of two cone detectors with orthogonal PS could drive the variation in PS and that feedback from horizontal cells to cones could explain the differential amplification of PS. This elegant arrangement, in which weak PS of cones is amplified and tuned by retinal networks, allows for PS without interfering with sampling of other visual information and illustrates how sensory systems may simultaneously process disparate aspects of physical environments.

What has driven the evolution of multiple cone classes in visual systems: object contrast enhancement or light flicker elimination?

BACKGROUND: Two competing theories have been advanced to explain the evolution of multiple cone classes in vertebrate eyes. These two theories have important, but different, implications for our understanding of the design and tuning of vertebrate visual systems. The 'contrast theory' proposes that multiple cone classes evolved in shallow-water fish to maximize the visual contrast of objects against diverse backgrounds. The competing 'flicker theory' states that multiple cone classes evolved to eliminate the light flicker inherent in shallow-water environments through antagonistic neural interactions, thereby enhancing object detection. However, the selective pressures that have driven the evolution of multiple cone classes remain largely obscure. RESULTS: We show that two critical assumptions of the flicker theory are violated. We found that the amplitude and temporal frequency of flicker vary over the visible spectrum, precluding its cancellation by simple antagonistic interactions between the output signals of cones. Moreover, we found that the temporal frequency of flicker matches the frequency where sensitivity is maximal in a wide range of fish taxa, suggesting that the flicker may actually enhance the detection of objects. Finally, using modeling of the chromatic contrast between fish pattern and background under flickering illumination, we found that the spectral sensitivity of cones in a cichlid focal species is optimally tuned to maximize the visual contrast between fish pattern and background, instead of to produce a flicker-free visual signal. CONCLUSIONS: The violation of its two critical assumptions substantially undermines support for the flicker theory as originally formulated. While this alone does not support the contrast theory, comparison of the contrast and flicker theories revealed that the visual system of our focal species was tuned as predicted by the contrast theory rather than by the flicker theory (or by some combination of the two). Thus, these findings challenge key assumptions of the flicker theory, leaving the contrast theory as the most parsimonious and tenable account of the evolution of multiple cone classes.

Modulation of environmental light alters reception and production of visual signals in Nile tilapia.

Signal reception and production form the basis of animal visual communication, and are largely constrained by environmental light. However, the role of environmental light in producing variation in either signal reception or production has not been fully investigated. To chart the effect of environmental light on visual sensitivity and body colouration throughout ontogeny, we measured spectral sensitivity, lens transmission and body pattern reflectance from juvenile and adult Nile tilapia held under two environmental light treatments. Spectral sensitivity in juveniles reared under a broad-spectrum light treatment and a red-shifted light treatment differed mostly at short wavelengths, where the irradiance of the two light treatments differed the most. In contrast, adults held under the same two light treatments did not differ in spectral sensitivity. Lens transmission in both juveniles and adults did not differ significantly between environmental light treatments, indicating that differences in spectral sensitivity of juveniles originated in the retina. Juveniles and adults held under the two environmental light treatments differed in spectral reflectance, and adults transferred to a third, white light treatment differed in spectral reflectance from their counterparts held under the two original treatments. These results demonstrate that environmental light plays a crucial role in shaping signal reception in juveniles and signal production throughout ontogeny, reinforcing the notion that environmental light has the capacity to influence animal communication, and suggesting that the characteristics of environmental light should be considered in models of ecological speciation.

High complexity of aquatic irradiance may have driven the evolution of four-dimensional colour vision in shallow-water fish.

Humans use three cone photoreceptor classes for colour vision, yet many birds, reptiles and shallow-water fish are tetrachromatic and use four cone classes. Screening pigments, which narrow the spectrum of photoreceptors in birds and diurnal reptiles, render visual systems with four cone classes more efficient. To date, however, the question of tetrachromacy in shallow-water fish that, like humans, lack screening pigments, is still unsolved. We raise the possibility that tetrachromacy in fish has evolved in response to higher spectral complexity of underwater light. We compared the dimensionality of colour vision in humans and fish by examining the spectral complexity of the colour signal reflected from objects into their eyes. We show that fish require four to six cone classes to reconstruct the colour signal of aquatic objects at the accuracy level achieved by humans viewing terrestrial objects. This is because environmental light, which alters the colour signals, is more complex and contains more spectral fluctuations underwater than on land. We further show that fish cones are better suited than human cones to detect these spectral fluctuations, suggesting that the capability of fish cones to detect high-frequency fluctuations in the colour signal confers an advantage. Taken together, we propose that tetrachromacy in fish has evolved to enhance the reconstruction of complex colour signals in shallow aquatic environments. Of course, shallow-water fish might possess fewer than four cone classes; however, this would come with the inevitable loss in accuracy of signal reconstruction.

Ontogeny in the visual system of Nile tilapia.

Retinal neurogenesis in fish facilitates cellular rearrangement throughout ontogeny, potentially allowing for optimization of the visual system to shifts in habitat and behaviour. To test this possibility, we studied the developmental trajectory of the photopic visual process in the Nile tilapia. We examined ontogenetic changes in lens transmission, photoreceptor sensitivity and post-receptoral sensitivity, and used these to estimate changes in cone pigment frequency and retinal circuitry. We observed an ontogenetic decrease in ultraviolet (UV) photoreceptor sensitivity, which resulted from a reduction in the SWS1 cone pigment frequency, and was associated with a reduction in lens transmission at UV wavelengths. Additionally, post-receptoral sensitivity to both UV and long wavelengths decreased with age, probably reflecting changes in photoreceptor sensitivity and retinal circuitry. This novel remodelling of retinal circuitry occurred following maturation of the visual system but prior to reaching adulthood, and thus may facilitate optimization of the visual system to the changing sensory demands. Interestingly, the changes in post-receptoral sensitivity to long wavelengths could not be predicted by the changes observed in lens transmission, cone pigment frequency or photoreceptor sensitivity. This finding emphasizes the importance of considering knowledge of visual sensitivity and retinal processing when studying visual adaptations and attempting to relate visual function to the natural environment. This study advances our understanding of ontogeny in visual systems and demonstrates that the association between different elements of the visual process can be explored effectively by examining visual function throughout ontogeny.

In ovo thyroxine exposure alters later UVS cone loss in juvenile rainbow trout.

Thyroid hormones (THs) play a vital role in vertebrate neural development, and, together with the beta isoform of the thyroid hormone receptor (TRß), the development and differentiation of cone photoreceptors in the vertebrate retina. Rainbow trout undergo a natural process of cone cell degeneration during development and this change in photoreceptor distribution is regulated by thyroxine (T4; a thyroid hormone). In an effort to further understand the role of T4 in photoreceptor ontogeny and later developmental changes in photoreceptor subtype distribution, the influence of enhanced in ovo T4 content on the onset of opsin expression and cone development was examined. Juvenile trout reared from the initial in ovo-treated embryos were challenged with exogenous T4 at the parr stage of development to determine if altered embryonic exposure to yolk THs would affect later T4-induced short-wavelength-sensitive (SWS1) opsin transcript downregulation and ultraviolet-sensitive (UVS) cone loss. In ovo TH manipulation led to upregulation of both SWS1 and long-wavelength-sensitive (LWS) opsin transcripts in the pre-hatch rainbow trout retina and to changes in the relative expression of TRß. After 7 days of exposure to T4, juveniles that were also treated with T4 in ovo had greatly reduced SWS1 expression levels and premature loss of UVS cones relative to T4-treated juveniles raised from untreated eggs. These results suggest that changes in egg TH levels can have significant consequences much later in development, particularly in the retina.

The underwater photic environment of Cape Maclear, Lake Malawi: comparison between rock- and sand-bottom habitats and implications for cichlid fish vision.

Lake Malawi boasts the highest diversity of freshwater fishes in the world. Nearshore sites are categorized according to their bottom substrate, rock or sand, and these habitats host divergent assemblages of cichlid fishes. Sexual selection driven by mate choice in cichlids led to spectacular diversification in male nuptial coloration. This suggests that the spectral radiance contrast of fish, the main determinant of visibility under water, plays a crucial role in cichlid visual communication. This study provides the first detailed description of underwater irradiance, radiance and beam attenuation at selected sites representing two major habitats in Lake Malawi. These quantities are essential for estimating radiance contrast and, thus, the constraints imposed on fish body coloration. Irradiance spectra in the sand habitat were shifted to longer wavelengths compared with those in the rock habitat. Beam attenuation in the sand habitat was higher than in the rock habitat. The effects of water depth, bottom depth and proximity to the lake bottom on radiometric quantities are discussed. The radiance contrast of targets exhibiting diffused and spectrally uniform reflectance depended on habitat type in deep water but not in shallow water. In deep water, radiance contrast of such targets was maximal at long wavelengths in the sand habitat and at short wavelengths in the rock habitat. Thus, to achieve conspicuousness, color patterns of rock- and sand-dwelling cichlids would be restricted to short and long wavelengths, respectively. This study provides a useful platform for the examination of cichlid visual communication.

Functional diversity in the color vision of cichlid fishes.

BACKGROUND: Color vision plays a critical role in visual behavior. An animal's capacity for color vision rests on the presence of differentially sensitive cone photoreceptors. Spectral sensitivity is a measure of the visual responsiveness of these cones at different light wavelengths. Four classes of cone pigments have been identified in vertebrates, but in teleost fishes, opsin genes have undergone gene duplication events and thus can produce a larger number of spectrally distinct cone pigments. In this study, we examine the question of large-scale variation in color vision with respect to individual, sex and species that may result from differential expression of cone pigments. Cichlid fishes are an excellent model system for examining variation in spectral sensitivity because they have seven distinct cone opsin genes that are differentially expressed. RESULTS: To examine the variation in the number of cones that participate in cichlid spectral sensitivity, we used whole organism electrophysiology, opsin gene expression and empirical modeling. Examination of over 100 spectral sensitivity curves from 34 individuals of three species revealed that (1) spectral sensitivity of individual cichlids was based on different subsets of four or five cone pigments, (2) spectral sensitivity was shaped by multiple cone interactions and (3) spectral sensitivity differed between species and correlated with foraging mode and the spectral reflectance of conspecifics. Our data also suggest that there may be significant differences in opsin gene expression between the sexes. CONCLUSIONS: Our study describes complex opponent and nonopponent cone interactions that represent the requisite neural processing for color vision. We present the first comprehensive evidence for pentachromatic color vision in vertebrates, which offers the potential for extraordinary spectral discrimination capabilities. We show that opsin gene expression in cichlids, and possibly also spectral sensitivity, may be sex-dependent. We argue that females and males sample their visual environment differently, providing a neural basis for sexually dimorphic visual behaviour. The diversification of spectral sensitivity likely contributes to sensory adaptations that enhance the contrast of transparent prey and the detection of optical signals from conspecifics, suggesting a role for both natural and sexual selection in tuning color vision.

Ultraviolet polarization vision and visually guided behavior in fishes.

Teleost fishes are capable of detecting and behaviorally responding to linearly polarized light. Fish exhibit free-swimming spatial orientation to imposed and natural polarized light fields, and the fidelity of this spatial orientation depends heavily on UV and short wavelength content of the polarization field. Fish make fine-scale behavioral discriminations between stimuli that differ in e-vector orientation, independent of brightness. The detection of polarized light by photoreceptors is based on specializations of the disk membrane in the outer segment of cones that permit preferential absorption of axial and transverse polarized light. Differential polarization detectors that have overlapping spectral sensitivity in the UV short wavelength spectrum mediate polarization sensitivity. These differential detectors are based on cone photoreceptors that share spectral sensitivity in the UV short wavelength spectrum: the alpha-band of UV-sensitive cone mechanism as the vertical detector, and the beta-band of mid- and long-wavelength sensitive cone mechanisms as the horizontal detector. Negative feedback of horizontal cells on cones govern opponent interactions between differentially sensitive polarization detectors. Polarization opponency functions to enhance e-vector contrast under conditions that vary in degree of polarization and ambient intensity. Ontogenetic changes in the cone mosaic, resulting from programmed cell death and regeneration of UV-sensitive cones, alter the retinal location of polarization sensitivity. These developmental changes greatly influence behavioral responses to polarized light.

Degeneration and regeneration of ultraviolet cone photoreceptors during development in rainbow trout.

Ultraviolet-sensitive (UVS) cones disappear from the retina of salmonid fishes during a metamorphosis that prepares them for deeper/marine waters. UVS cones subsequently reappear in the retina near sexual maturation and the return migration to natal streams. Cellular mechanisms of this UVS cone ontogeny were investigated using electroretinograms, in situ hybridization, and immunohistochemistry against opsins during and after thyroid hormone (TH) treatments of rainbow trout (Oncorhynchus mykiss). Increasing TH levels led to UVS cone degeneration. Labeling demonstrated that UVS cone degeneration occurs via programmed cell death and caspase inhibitors can inhibit this death. After the cessation of TH treatment, UVS cones regenerated in the retina. Bromodeoxyuridine (BrdU) was applied after the termination of TH treatment and was detected in the nuclei of cells expressing UVS opsin. BrdU was found in UVS cones but not other cone types. The most parsimonious explanation for the data is that UVS cones degenerated and UVS cones were regenerated from intrinsic retinal progenitor cells. Regenerating UVS cones were functionally integrated such that they were able to elicit electrical responses from second-order neurons. This is the first report of cones regenerating during natural development. Both the death and regeneration of cones in retinae represent novel mechanisms for tuning visual systems to new visual tasks or environments.

Proteomic analysis of opsins and thyroid hormone-induced retinal development using isotope-coded affinity tags (ICAT) and mass spectrometry.

PURPOSE: Analyses that reveal the relative abundance of proteins are informative in elucidating mechanisms of retinal development and disease progression. However, popular high-throughput proteomic methods do not reliably detect opsin protein abundance, which serve as markers of photoreceptor differentiation. We utilized thyroid-hormone (TH) treatment of rainbow trout (Oncorhynchus mykiss) as a model of cone apoptosis and cone regeneration. We used this model to investigate if emerging proteomic technology allows effective analysis of retinal development and opsin protein abundance. We also sought to begin a characterization of proteomic changes in the retina occurring with TH treatment and address whether TH affects proliferation or photoreceptor differentiation. METHODS: Retinal homogenates were prepared from control and TH-treated fish. Peptides from control and treated homogenates were differentially labeled, using isotope-code affinity tags (ICAT) and analyzed using capillary liquid chromatography-electrospray ionization-tandem mass spectrometry (capLC-ESI-MS/MS). This method identifies proteins and quantifies their relative abundance between two samples. RESULTS: The relative abundance of many retinal proteins changed during TH treatment. These included proteins from every functional class. We detected 1,684 different peptides, and our quantification suggests that 94 increased and 146 decreased in abundance more than 50% during TH treatment. Cell-cycle proteins appear to be increased, consistent with TH-inducing cell proliferation, similar to its effect in Xenopus. Other proteins associated with retinal development, such as deltaA and tubulins, changed in abundance during TH treatment. Rod opsin and three cone opsins were identified and the relative abundance of each changed with TH treatment. CONCLUSIONS: ICAT and capLC-ESI-MS/MS are an effective complement to other molecular approaches that investigate the mechanisms of retinal development. Unlike other proteomic techniques, this approach does not require development of species- or tissue-specific methodology, such as characterizing two dimensional (2D) gels or antibodies, in order to be practical as a high-throughput approach. Importantly, this technology was able to assess the relative abundance of opsin proteins. These findings represent the first high-throughput proteomic analysis of the retina and demonstrate the technique's ability to provide useful information in retinal development.

Ontogeny of ultraviolet-sensitive cones in the retina of rainbow trout (Oncorhynchus mykiss).

In order to facilitate emerging models of retinal development, we developed electroretinogram and in situ hybridization protocols to examine the ontogeny of photoreceptors in the retina of a land-locked salmonid, the rainbow trout (Oncorhynchus mykiss). We cloned cDNA fragments corresponding to the rod opsin and each of the four cone opsin gene families, which we utilized to produce riboprobes. We established the specificity of the in situ hybridization protocol by examining subcellular signal localization and through double-labeling experiments. We confirm the assumption that the accessory corner cones in the square mosaic are the ultraviolet wavelength-sensitive (UVS) cone photoreceptor (i.e., they express an SWS1 opsin) and observed UVS cones throughout the retina of small trout. Larger fish have a decrease in sensitivity to short wavelength light stimuli and the distribution of UVS cones in the mature retina is limited to the dorsal-temporal quadrant. These larger fish also possess differentiated UVS cones in the peripheral germinal zone (PGZ), including within areas peripheral to mature retina lacking UVS cones. These data are consistent with the loss of putative UVS cones from the PGZ of a migratory salmonid of another genus, and thus the disappearance of UVS cones appears to be general to the Family Salmonidae, regardless of life history strategy. The generation, differentiation, and subsequent loss of UVS cones in the smolt PGZ is a dramatic example of the supposition that the mechanisms of PGZ development recapitulate the retinal embryogenesis of that species.

Identification of a unique transcript down-regulated in the retina of rainbow trout (Oncorhynchus mykiss) at smoltification.

Developmental and physiological changes in the retina of salmonid fishes occur during smoltification, a metamorphic event associated with thyroid hormone that prepares salmon for oceanic migration. These changes include loss of ultraviolet-sensitive (UVS) cone photoreceptors, switching of visual pigments, alterations in thyroid hormone regulation, and associated changes in behavior. This model provides an opportunity to study substantial neuronal development within an established retina. Little is known, however, about how higher order neurons are altered or how retinal gene expression changes during this transition. Here, we have used differential display RT-PCR and RACE-PCR to identify a previously uncharacterized gene transcript in Oncorhynchus mykiss under developmental regulation in the retina during smoltification, rtp12.5. This unique cDNA encodes a putative protein 112 amino acids long similar to a hypothetical human open reading frame located on chromosome 14.Q24.2. Differential expression was confirmed by RNA dot blot and in situ hybridization. We also present O. mykiss sep15 cDNA sequence and describe its expression in the vertebrate retina. Considering the expression pattern within retinal tissue observed by in situ hybridization, rtp12.5 may be under TH regulation and involved in neuronal remodeling of the retina during loss of UVS cones.

Ontogeny of melanophore photosensitivity in rainbow trout (Oncorhynchus mykiss).

Migratory species experience morphological and physiological changes during transitions between different life stages. In particular, modification of sensory systems is critical for animals to adapt to new environments. For example, to prepare for entry into seawater, salmonids undergo smoltification with dramatic changes in ultraviolet photoreceptors and polarized vision, which are important for orientation and foraging behaviours. Extraretinal organs are also involved in photoreception; however, the ontogenetic development of extraretinal photoreceptors is not well known, especially in migratory species. Here, we investigated whether rainbow trout dermal photoreceptors, melanophores, undergo change in spectral sensitivity during smoltification and which candidate molecules may account for this ontogenetic alteration. Our results showed that, contrary to parr melanophores which are insensitive to light, smolt melanophores displayed chromatic photoresponses with the emergence of cryptochrome and melanopsin expression. We suggest that these modifications may benefit the active foraging behaviour of smolts and enable adaptation to variable environments.

The influence of chromatic background on the photosensitivity of tilapia erythrophores.

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.

Possible involvement of cone opsins in distinct photoresponses of intrinsically photosensitive dermal chromatophores in tilapia Oreochromis niloticus.

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.

Feedback from horizontal cells to cones mediates color induction and may facilitate color constancy in rainbow trout.

Color vision is most beneficial when the visual system is color constant and can correct the excitations of photoreceptors for differences in environmental irradiance. A phenomenon related to color constancy is color induction, where the color of an object shifts away from the color of its surroundings. These two phenomena depend on chromatic spatial integration, which was suggested to originate at the feedback synapse from horizontal cells (HC) to cones. However, the exact retinal site was never determined. Using the electroretinogram and compound action potential recordings, we estimated the spectral sensitivity of the photoresponse of cones, the output of cones, and the optic nerve in rainbow trout. Recordings were performed before and following pharmacological inhibition of HC-cone feedback, and were repeated under two colored backgrounds to estimate the efficiency of color induction. No color induction could be detected in the photoresponse of cones. However, the efficiency of color induction in the cone output and optic nerve was substantial, with the efficiency in the optic nerve being significantly higher than in the cone output. We found that the efficiency of color induction in the cone output and optic nerve decreased significantly with the inhibition of HC-cone feedback. Therefore, our findings suggest not only that color induction originates as a result of HC-cone feedback, but also that this effect of HC-cone feedback is further amplified at downstream retinal elements, possibly through feedback mechanisms at the inner plexiform layer. This study provides evidence for an important role of HC-cone feedback in mediating color induction, and therefore, likely also in mediating color constancy.

Spectral sensitivity of single cones in rainbow trout (Oncorhynchus mykiss): a whole-cell voltage clamp study.

The UVS cone mechanism is known to light adapt at low intensities in comparison to other cones. We were interested in whether this property was related to higher sensitivity in UVS cones or to network adjustments in sensitivity. We investigated spectral sensitivity of 107 individual cone photoreceptors in rainbow trout (Oncorhynchus mykiss) using a whole-cell voltage clamp technique. Mean time-to-peak response was 339+/-90ms and flash sensitivity for a 100ms flash was 4.37x10(-3)+/-2.50x10(-3)pAphotons(-1)microm(2), with no significant differences between the UVS, SWS, MWS and LWS cone classes. The spectral sensitivity of the UVS, SWS and LWS cones conformed to the expression of SWS1, SWS2 and LWS opsin genes. The spectral sensitivity of MWS cones, however, showed clear evidence of co-expression of RH2a and RH2b opsin pigments. The fish used in this study bridged the ontogenetic stage where the MWS cones shift their expression from RH2b to RH2a.

Retinal processing and opponent mechanisms mediating ultraviolet polarization sensitivity in rainbow trout (Oncorhynchus mykiss).

A number of teleost fishes have photoreceptor mechanisms to detect linearly polarized light. We studied the neuronal mechanism underlying this ability. It was found that a polarized signal could be detected in rainbow trout (Oncorhynchus mykiss) both in the electroretinogram (ERG) and in the compound action potential (CAP) measured in the optic nerve, indicating a strong retinal contribution to the processing of polarized light. The CAP recordings showed a W-shaped sensitivity curve, with a peak at 0 degrees , 90 degrees and 180 degrees , consistent with processes for both vertical and horizontal orientation. By contrast, the ERG recordings reveal a more complex pattern. In addition to the peaks at 0 degrees , 90 degrees and 180 degrees , two additional peaks appeared at 45 degrees and 135 degrees . This result suggests a specialized contribution of the outer retina in the processing of polarized light. The spectral sensitivity of the mechanisms responsible for these intermediate peaks was studied using chromatic adaptation. Here we show that long wavelength-sensitive (LWS) cone mechanism adaptation shifted the intermediate peaks towards 90 degrees , whereas ultraviolet-sensitive (UVS) cone mechanism adaptation shifted the peaks away from 90 degrees towards either 0 degrees or 180 degrees . These results provide further confirmation that the 90 degrees peak is dominated by the LWS cone mechanism and the 0 degrees and 180 degrees peaks are dominated by the UVS cone mechanism. In addition, a pharmacological approach was used to examine the retinal neural mechanisms underlying polarization sensitivity. The effect of blocking negative feedback from horizontal cells to cones on the ERG was studied by making intraocular injections of low doses of cobalt, known to block this feedback pathway. It was found that the intermediate peaks seen in the ERG polarization sensitivity curves were eliminated after application of cobalt, suggesting that these peaks are due to outer retinal inhibition derived from feedback of horizontal cells onto cones. A simple computational model was developed to evaluate these results. The model consists of opponent and non-opponent processing elements for the two polarization detectors. This model provides a first approximation analysis suggesting that opponent processing occurs in the outer retina for polarization vision. Although it seems that polarization vision uses a slightly more complicated coding scheme than colour vision, the results presented in this paper suggest that opponent and non-opponent channels process polarization information.