Shortwave-sensitive Cones Research Articles

On the retinae of Glis and Graphiurus: photoreceptor and ganglion cell populations, an absence of shortwave-sensitive cones, and some other features (Rodentia: Gliridae)

The retina of the fat dormouse Glis glis was studied histologically. Opsin immunolabeling identified an unusually dense population of rod photoreceptors (ca. 600,000–780,000/mm²) and a low-density population of L cone photoreceptors containing the longwave-sensitive (LWS) cone opsin, with a shallow maximum of ca. 3,300/mm² in temporal retina and ca. 2,000–3,000/mm² in the remaining retina. Hence the cones comprise only 0.25–0.50% of the photoreceptors. There are no S cones expressing the shortwave-sensitive (SWS1) opsin that is the second cone opsin in most mammals, conveying dichromatic colour vision. We conclude that the fat dormouse is a cone monochromat. Sequencing of the tuning-relevant exon 1 of the SWS1 opsin gene revealed that the gene would have coded for a UV-sensitive visual pigment, but that it contains mutational changes making it nonfunctional. Retinal interneurons (rod bipolar cells, horizontal cells, several amacrine cell types) have rodent-typical features. NeuN-labeled presumed retinal ganglion cells have densities between ca. 4000/mm² in temporal central retina and 850–1300/mm² in peripheral retina. The peak ganglion cell density would result in an estimated visual acuity maximum of ca. 1.8 cycles/degree or ca. 33 minutes of arc. Assessment of a further Gliridae species, the African dormouse Graphiurus sp., also revealed a high rod density, low L cone density, and an absence of SWS1 opsin.

Complementary shifts in photoreceptor spectral tuning unlock the full adaptive potential of ultraviolet vision in birds.

Color vision in birds is mediated by four types of cone photoreceptors whose maximal sensitivities (λmax) are evenly spaced across the light spectrum. In the course of avian evolution, the λmax of the most shortwave-sensitive cone, SWS1, has switched between violet (λmax > 400 nm) and ultraviolet (λmax < 380 nm) multiple times. This shift of the SWS1 opsin is accompanied by a corresponding short-wavelength shift in the spectrally adjacent SWS2 cone. Here, we show that SWS2 cone spectral tuning is mediated by modulating the ratio of two apocarotenoids, galloxanthin and 11’,12’-dihydrogalloxanthin, which act as intracellular spectral filters in this cell type. We propose an enzymatic pathway that mediates the differential production of these apocarotenoids in the avian retina, and we use color vision modeling to demonstrate how correlated evolution of spectral tuning is necessary to achieve even sampling of the light spectrum and thereby maintain near-optimal color discrimination.

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

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.

Cone fundamentals: A model for the future of colorimetry

Cone fundamentals is the name given to the relative spectral sensitivity of long-wave sensitive, medium-wave sensitive and short-wave sensitive cones. Cone fundamentals represent the spectral absorptances of the retinal photopigments in the cones multiplied by the spectral transmittances of the ocular media through which light passes to reach the retina. The CIE 2006 cone fundamentals are based on the Stiles and Burch experimental colour matches and have been validated by recent microspectrophotometric measurements and modelling. A new physiologically based version of the MacLeod–Boynton chromaticity diagram is proposed by the CIE as well as a cone-fundamental-based ( xF, yF) chromaticity diagram. All the data necessary to use these concepts for high-precision colorimetric calculations are readily available.

Characterization of retinoic acid–induced neurobehavioral effects in developing zebrafish

Retinoic signaling plays an important role in cell proliferation and differentiation. Disruption of retinoic signaling via excessive or deficient retinoic acid can cause teratogenic effects on developing embryos. Similar to retinoic acid, many xenobiotic environmental pollutants have been found to disrupt retinoic signaling through binding and eliciting agonistic activity on retinoic acid receptors. Currently, studies of retinoic acid or retinoic acid-like compounds in aquatic organisms have mainly focused on teratogenicity and few studies have explored their neurobehavioral toxicity. In the present study, the authors used retinoic acid as an example to explore the neurobehavioral toxicity associated with developmental exposure of retinoic acid-like compounds in zebrafish. The findings confirmed retinoic acid's teratogenic effects such as bent spine, malformed tail, and pericardial edema in developing zebrafish with a median effective concentration of 2.47 nM. Retinoic acid-induced cell apoptosis at 24 h postfertilization was consistently found in the eye and tail regions of embryos. Spontaneous movement as characterized by tail bend frequency was significantly increased in zebrafish embryos following exposure to 2 nM and 8 nM retinoic acid. Relatively low-dose retinoic acid exposure of 2 nM led to fast locomotion behavior in the dark period and hyperactivity during light-dark photoperiod stimulation. The 2-nM retinoic acid exposure also led to alterations of neurobehavior- and optic nerve-related genes, with the transforming growth factor-β signal transduction inhibitor noggin (nog) and the spinal cord marker homeobox c3a (hox) being underexpressed and the retinal G protein-coupled receptor a (rgr), the photoreceptor cell marker rhodopsin (rho), and the short wave-sensitive cone pigment opsin 1 (opn1sw1) being overexpressed. Increased expression of opn1sw1 and rho was confirmed by whole-mount in situ hybridization. Whether the misexpression of these genes leads to the neurobehavioral changes merits further study. The findings demonstrated that low-dose retinoic acid exposure perturbed the visual system and optic nerve development and caused hyperactivity in developing zebrafish.

Which Visual Pathways Cause Fixation-Related Inhibition?

Visual stimuli can both inhibit and activate motor mechanisms. In one well-known example, the latency of saccadic eye movements is prolonged in the presence of a fixation stimulus, relative to the case in which the fixation stimulus disappears before the target appears. This automatic sensory-motor effect, known as the gap effect or fixation-offset effect, has been associated with inhibitory connections within the superior colliculus (SC). Visual information is provided to the SC and other oculomotor areas, such as the frontal eye fields (FEF), mainly by the magnocellular geniculostriate pathway, and also by the retinotectal pathway. We tested whether signals in these pathways are necessary to create fixation-related inhibition, by using stimuli invisible to them. We found that such stimuli, visible only to short-wave-sensitive cones (S cones), do produce fixation-related inhibition (including when warning effects were equated). We also demonstrate that this fixation-related inhibition cannot be explained by residual activation of luminance pathways and must be caused by a route separate from that of luminance fixation signals. Thus there are at least two routes that cause fixation-related inhibition, and direct sensory input to the SC or FEF by the magnocellular or retinotectal pathways is not required. We discuss the implications that there may be both cortical and collicular mechanisms.

Spectral tuning and evolution of short wave-sensitive cone pigments in cottoid fish from Lake Baikal.

The cottoid fishes of Lake Baikal in eastern Siberia provide a unique opportunity to study the evolution of visual pigments in a group of closely related species exposed to different photic environments. Members of this species flock are adapted to different depth habitats down to >1000 m, and both the rod and cone visual pigments display short wave shifts as depth increases. The blue-sensitive cone pigments of the SWS2 class cluster into two species groups with lambda(max) values of 450 and 430 nm, with the pigment in Cottus gobio, a cottoid fish native to Britain, forming a third group with a lambda(max) of 467 nm. The sequences of the SWS2 opsin gene from C. gobio and from two representatives of the 450 and 430 nm Baikal groups are presented. Approximately 6 nm of the spectral difference between C. gobio and the 450 nm Baikal group can be ascribed to the presence of a porphyropsin/rhodopin mixture in C. gobio. Subsequent analysis of amino acid substitutions by site-directed mutagenesis demonstrates that the remainder of the shift from 461 to 450 nm arises from a Thr269Ala substitution and the shift from 450 to 430 nm at least partly from Thr118Ala and Thr118Gly substitutions. The underlying adaptive significance of these substitutions in terms of spectral tuning and signal-to-noise ratio is discussed.

Cone inputs to murine retinal ganglion cells

Responses of single retinal ganglion cells in different areas of mouse retina were studied to determine their cone inputs, using spectral sensitivity functions and chromatic adaptation. Spectral sensitivity curves were based on threshold response criteria to full field stimulation. The retina of the mouse was viewed through a dilated pupil with a surgical microscope. Ganglion cells were classified into three groups: one receiving inputs from short wave sensitive cones, a second receiving inputs from only middle wavelength sensitive cones and a third receiving inputs from both of these types of cones. The ventral retina, contained a large fraction of the first group of ganglion cells. The dorsal retina and the border between these two areas contained relatively more of the latter two groups. A small fraction of cells were found which displayed antagonistic-like interactions between photoreceptor systems. The results demonstrate that single ganglion cells in mouse retina can select responses from only one of the two cone mechanisms present in this retina, even in areas containing both types of cones.

Localization of nitric oxide synthase in the tree shrew retina.

Nitric oxide (NO) is a novel neuronal messenger that likely influences retinal function by activating retinal guanylyl cyclase to increase levels of cGMP. In the present study, the localization of neuronal nitric oxide synthase (nNOS, Type I NOS) in the cone-dominant tree shrew retina was studied using NADPH-d histochemistry and nNOS immunocytochemistry. Both NADPH-d and nNOS-immunoreactivity (IR) labeled the inner segments of rods and the myoids of a regular subpopulation of cones, with their corresponding nuclei outlined. The labeled cone myoids were co-localized with a marker for short-wave-sensitive (SWS) cones (S-antigen) and also displayed the regular triangular packing and density (7%) characteristic of SWS cones in tree shrew and other mammalian retinas. These measures confirmed the identity of the labeled cones as SWS cones. Photoreceptor ellipsoids of all cones were strongly labeled by NADPH-d reactivity, but lacked nNOS-IR. Another novel finding in tree shrew retina was that both NADPH-d and nNOS-IR labeled Muller cells, which have not been labeled by nNOS-IR in other mammalian retinas. Consistent with findings in rod-dominant retinas, two types of amacrine cells at the vitreal edge of the inner nuclear layer and a subpopulation of displaced amacrine cells at the scleral edge of the ganglion cell layer were labeled by both NADPH-d and nNOS-IR. Processes of these labeled cells were seen to extend into the inner plexiform layer, where dense punctate label was seen, especially in the central sublamina. These results show that localization of NOS in the cone-dominant tree shrew retina shares some common properties with rod-dominant mammalian retinas, but also shows some species-specific characteristics. The new finding of nNOS localization in tree shrew SWS cones and rods, but not in other cones, raises interesting questions about the roles of NO in the earliest level of visual processing.

Decline of circadian photosensitivity associated with retinal degeneration in CBA/J- rd/rd mice

Although it is well known that visual image formation is mediated by rods and cones, photoreceptors implicated in entrainment of circadian rhythms remain unknown. In our previous study, 12-week-old CBA/J- rd/rd mice were found to show decreased sensitivity in circadian photoreception. Because rd gene induces different age-related degeneration of cones and rods, we have examined age-related changes in the sensitivity to a light pulse for phase-shifting the locomotor activity rhythms in CBA/J- rd/rd mice. Since mice have middlewave-sensitive cones (M-cone, 510 nm) and shortwave-sensitive cones (S-cone, 360 nm), a 15-min monochromatic light pulse (501.6 or 359 nm; 10 μW cm −2 s −1) was given at circadian time (CT) 16, 2 weeks after being transferred to constant darkness (DD) and the magnitude of phase shift was determined. The amount of phase shift by a green light pulse (501.6 nm) gradually declined from 8 to 16 weeks after birth and the similar tendency was also observed on a UV pulse (359 nm). The similar pattern of age-related decline in the number of cones and circadian photosensitivity in CBA/J- rd/rd mice suggest that cones are involved in the circadian photoreception in mice.

Distribution of cone photoreceptors in the mammalian retina.

The retina of mammals contains various amounts of cone photoreceptors that are relatively evenly distributed and display a radially or horizontally oriented area of peak density. In most mammalian species two spectrally different classes of cone can be distinguished with various histochemical and physiological methods. These cone classes occur in a relatively constant ratio, middle-to-longwave sensitive cones being predominant over short-wave cones. Recent observations do not support the idea that each cone subpopulation is uniformly distributed across the retina. With appropriate type-specific markers, unexpected patterns of colour cone topography have been revealed in certain species. In the mouse and the rabbit, the "standard" uniform pattern was found to be confined exclusively to the dorsal retina. In a ventral zone of variable width all cones express short-wave pigment, a phenomenon whose biological significance is not known yet. Dorso-ventral asymmetries have been described in lower vertebrates, matching the spectral distribution of light reaching the retina from various sectors of the visual field. It is not clear, however, whether the retinal cone fields in mammals carry out a function similar to that of their counterparts in fish and amphibians. Since in a number of mammalian species short-wave cones are the first to differentiate, and the expression of the short-wave pigment seems to be the default pathway of cone differentiation, we suggest that the short-wave sensitive cone fields are rudimentary areas conserving an ancestral stage of the photopigment evolution.

Colour vision in the passeriform bird, Leiothrix lutea: correlation of visual pigment absorbance and oil droplet transmission with spectral sensitivity

The visual receptors in the retina of the passeriform bird Leiothrix lutea were examined microspectro-photometrically. The rods had a maximum absorbance close to 500 nm. Four spectrally different classes of single cone were identified with typical combinations of photopigments and oil droplets: a long-wave sensitive cone with a photopigment P568 and a droplet with a cut-off wavelength at 564 nm, a middle-wave sensitive cone with a P499 and a droplet with a cut-off at 506 nm, a short-wave sensitive cone with a P454 and a droplet with maximum absorbance below 410nm and an ultraviolet sensitive cone with a P355 and a transparent droplet. Double cones possessed a P568 in both the principal and accessory members. A pale droplet with variable absorbance (maximal at about 420 nm) was associated with the principal member whereas the ellipsoid region of the accessory member contained only low concentrations of carotenoid. The effective spectral sensitivities of the different cone classes were calculated from the characteristic combinations of oil droplets and photopigments and corrected for the absorbance of the ocular media. Comparison of these results with the behavioural spectral sensitivity function of Leiothrix lutea suggests that the increment threshold photopic spectral sensitivity of this avian species is mediated by the 4 single cone classes modified by neural opponent mechanisms.

A locus control region adjacent to the human red and green visual pigment genes

Deletion of sequences 5′ of the human red and green pigment gene array results in blue cone monochromacy, a disorder in which both red and green cone function are absent. To test whether these sequences are required for transcription of the adjacent visual pigment genes in cone photoreceptors, we produced transgenic mice carrying sequences upstream of the red and green pigment genes fused to a β-galactosidase reporter. The patterns of transgene expression indicate that the human sequences direct expression to both long and short wave-sensitive cones in the mouse retina and that a region between 3.1 kb and 3.7 kb 5′ of the red pigment gene transcription initiation site is essential for expression. Sequences within this region are highly conserved among humans, mice, and cattle, even though the latter two species have only a single visual pigment gene at this locus. These experiments suggest a model in which an interaction between the conserved 5′ region and either the red or the green pigment gene promoter determines which of the two genes a given cone expresses.

Hering's opponent colour channels do not exist in the primate retinogeniculate pathway.

We have used pure hue contrast stimuli to examine the responses of single primate retinal ganglion cells. Only one subset of these cells, those excited by the short wave sensitive cones and inhibited by the other cones, i.e. the blue-on/yellow-off variety, can distinguish white/yellow contrast. This implies that the paired opponent channels of Hering 's colour vision theory do not exist in the retinogeniculate pathway.

The saturation of monochromatic lights obliquely incident on the retina.

Foveal dark-adaptation undertaken to test the hypothesis that the excitation of rods causes the desaturation of 'yellow' lights in a 1 degree field traversing the margin of the pupil, fails to exclude that possibility. The desaturation is largest for a 1 degree outside diameter annular test, is still measurable with a 0.5 degree circular disk, but disappears for a 0.29 degree disk. The supersaturation of obliquely incident 501.2 nm test light follows the opposite pattern; it disappears with an annulus and is largest for a 0.29 degree circular field. It is unlikely that rods replace short-wave sensitive cones in the trichromatic match of an obliquely incident test with normally incident primaries. If rods as well as all three cones species are involved, the matches might not be trichromatic in the strong sense. Grassmann's law of scalar multiplication was tested and shown not to hold for the match of an obliquely incident test with normally incident primaries, though it remains valid whenever, both primaries and test strike the retina at the same angle of incidence (independent of that angle). The result in section 3 (above) cannot be due to rod intrusion. It persists (and becomes more conspicuous) on backgrounds (4.0 log scotopic td) which saturate rods. Moreover obliquely incident 'yellow' lights remain desaturated in intervals in the dark after a full bleach, whilst the test field is below rod threshold. The amount of desaturation does not differ appreciably from that normally found. The assumption of the unified theory of Alpern, Kitahara & Tamaki (1983) that the outer segments of only a single set of three cone species (with acceptance angles wide enough to include the entire exit pupil) contain the visual pigments absorbing both the normally incident primaries and the obliquely incident test is disproved by these results. Failure of Grassmann's law is most conspicuous under the conditions for which the changes in saturation upon changing from normal to oblique incidence are greatest and least when the saturation changes are the smallest. Either all unified theories of the Stiles-Crawford effects are wrong or all the effects of oblique incidence operate at a stage in the visual process at which the effects of radiation of different wave-lengths are no longer compounded by the simple linear laws.

Retinal site of transient tritanopia.

WHEN the eye is adapted to a bright yellow light, a marked loss of sensitivity for short-wavelength stimuli occurs in the first few seconds after extinction of this light. This phenomenon was first reported by Stiles1 and has been termed transient tritanopia by Mollon and Polden2. The latter authors suggest that this apparent suppression of the short-wave sensitive cones (hereafter called blue cones) during rapid dark adaptation is effected through inhibition by the recovering long- and middle-wave sensitive cone mechanisms2 (hereafter called red and green cones, respectively). In this study we attempted to localise the site of blue cone inhibition through intra-retinal recordings from the intact rhesus monkey eye. We found transient tritanopia to be absent at the receptor level but present at the b-wave level, and we conclude that transient tritanopia originates at a site between the receptors and the bipolar cells, probably at the horizontal cells, as was earlier hypothesised by Augenstein and Pugh3.

The wavelength variation of the directional sensitivity of the stiles π 1(μ)

The variation of the directional sensitivity of Stiles' (1953) π 1(μ) mechanism was measured throughout the spectrum on one deuteranope. For wavelengths greater than about 550 nm, the directional properties measured for π 1(μ) are nearly identical to those for this deuteranope's π 5(μ) mechanism. This supports the hypothesis of Pugh (1976) that π 1(μ) is a composite mechanism so that in the yellow-red part of the spectrum its sensitivity is determined by long- (and/or medium-) wave sensitive cones. It also obviates the contradiction between the single π 1(μ) directional sensitivity result of Stiles (1939) and the suggestion ( Enoch and Stiles, 1961) that short-wave sensitive cones are more sensitive to light entering through the edge than through the center of the pupil. This deuteranope's long-wave sensitive cones appear to be most sensitive to backgrounds entering the eye 0.5 mm nasal, to the center of his pupil while his short-wave sensitive cones appear to be most sensitive to light entering 0.2 mm temporal to the center of the pupil. Differences in the estimate of losses in the lens below 450 nm in the spectrum are too large to exclude either self-screening or the Snyder-Pask waveguide theories as applied to this subject's short-wave sensitive cones.

Variation in the action spectrum of erythrolabe among deuteranopes.

1. Eight deuteranopes matched a mixture of a monochromatic light on the long wave side of the neutral point and a violet (450 nm) primary to a fixed white as well as a monochromatic light on the short wave side of the neutral point mixed with a red (650 nm) primary, to the same white. For lambda greater than 530 nm, the former set of matches defined the action spectrum of the long wave sensitive foveal cones, and for lambda less than 480 nm, the latter that of the short wave sensitive cones. 2. Individual differences in the former matches were approximately correlated with the respective ratio of the sensitivities of the wave-length of the anomaloscope primaries, in a way that individual differences of the latter were not. 3. Assuming that eye media differences alone account for the differences in long wave sensitive foveal action spectra, the spectral reflectivity of the foveal fundus was predicted for these deuteranopes. The prediction is inconsistent with measurement. 4. Thirteen deuteranopes matched monochromatic spectral lights with a green (535 nm) and a blue (460 nm) primary. The result were analysed by von Kries' method in which differences in matching due to differences in eye media absorption are obviated. The matches of five differed significantly from one another when so analysed. It was concluded that at least one of two action spectra of the foveal cones of every one of these five differed from that of all of the others. 5. The canon that deuteranopes accept normal colour matches was evaluated by confronting a single normal with five deuternopes in the analytical anomaloscope of Baker & Rushton, set in the mode of each of the five in turn. Obvious differences existed between this normal's matches and those of four of five deuteranopes. 6. Explanations for differences in the spectrum of erythrolabe in different deuteranopes are evaluated. The possibilities that all have the identical visual pigment but (a) in cones with different optical funnelling properties or (b) in different optical densities are considered. Preliminary results are not in agreement with the expectations of either of these ideas. 7. It is suggested that the visual pigment in the foveal long wave sensitive cones of different deuternopes (and of different normals) may have different extinction spectra. The idea is consistent with micro-spectrophotometric measurements of rhodopsin in individual rods from different frogs (Bowmaker, Loew & Leibman, 1975).