Assessing the use of genomic DNA as a predictor of the maximum absorbance wavelength of avian SWS1 opsin visual pigments

Recently, in vitro mutation studies have made it possible to predict the wavelengths of maximum absorbance (λ_max) of avian UV/violet sensitive visual pigments (SWS1) from the identity of a few key amino acid residues in the opsin gene. Given that the absorbance spectrum of a cone’s visual pigment and of its pigmented oil droplet can be predicted from just the λ_max, it may become possible to predict the entire spectral sensitivity of a bird using genetic samples from live birds or museum specimens. However, whilst this concept is attractive, it must be validated to assess the reliability of the predictions of λ_max from opsin amino acid sequences. In this paper, we have obtained partial sequences covering three of the known spectral tuning sites in the SWS1 opsin and predicted λ_max of all bird species for which the spectral absorbance has been measured using microspectrophotometry. Our results validate the use of molecular data from genomic DNA to predict the gross differences in λ_max between the violet- and ultraviolet-sensitive subtypes of SWS1 opsin. Additionally, we demonstrate that a bird, the bobolink Dolichonyx oryzivorus L., can have more than one SWS1 visual pigment in its retina.     

The photoactivation energy of the visual pigment in two spectrally different populations of <Emphasis Type="Italic">Mysis relicta</Emphasis> (Crustacea,Mysida)

We report the first study of the relation between the wavelength of maximum absorbance (λ_max) and the photoactivation energy (E _a) in invertebrate visual pigments. Two populations of the opossum shrimp Mysis relicta were compared. The two have been separated for 9,000 years and have adapted to different spectral environments (“Sea” and “Lake”) with porphyropsins peaking at λ_max=529 nm and 554 nm, respectively. The estimation of E _a was based on measurement of temperature effects on the spectral sensitivity of the eye. In accordance with theory (Stiles in Transactions of the optical convention of the worshipful company of spectacle makers. Spectacle Makers’ Co., London, 1948), relative sensitivity to long wavelengths increased with rising temperature. The estimates calculated from this effect are E _a,529=47.8±1.8 kcal/mol and E _a,554=41.5±0.7 kcal/mol (different at P<0.01). Thus the red-shift of λ_max in the “Lake” population, correlating with the long-wavelength dominated light environment, is achieved by changes in the opsin that decrease the energy gap between the ground state and the first excited state of the chromophore. We propose that this will carry a cost in terms of increased thermal noise, and that evolutionary adaptation of the visual pigment to the light environment is directed towards maximizing the signal-to-noise ratio rather than the quantum catch.     

Visual pigments of marine carnivores: pinnipeds, polar bear, and sea otter

Rod and cone visual pigments of 11 marine carnivores were evaluated. Rod, middle/long-wavelength sensitive (M/L) cone, and short-wavelength sensitive (S) cone opsin (if present) sequences were obtained from retinal mRNA. Spectral sensitivity was inferred through evaluation of known spectral tuning residues. The rod pigments of all but one of the pinnipeds were similar to those of the sea otter, polar bear, and most other terrestrial carnivores with spectral peak sensitivities (λ_max) of 499 or 501 nm. Similarly, the M/L cone pigments of the pinnipeds, polar bear, and otter had inferred λ_max of 545 to 560 nm. Only the rod opsin sequence of the elephant seal had sensitivity characteristic of adaptation for vision in the marine environment, with an inferred λ_max of 487 nm. No evidence of S cones was found for any of the pinnipeds. The polar bear and otter had S cones with inferred λ_max of ∼440 nm. Flicker-photometric ERG was additionally used to examine the in situ sensitivities of three species of pinniped. Despite the use of conditions previously shown to evoke cone responses in other mammals, no cone responses could be elicited from any of these pinnipeds. Rod photoreceptor responses for all three species were as predicted by the genetic data.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Spectral and polarized light sensitivity of photoreceptors in the compound eye of the cricket (Gryllus bimaculatus)

Summary Retinula cells in the compound eye of the cricket (Gryllus bimaculatus) were recorded intracellularly and stained with Lucifer yellow. Two different methods were used to determine the spectral sensitivity of these cells: a) the spectral scanning method, and b) the conventional flash method. Three spectral types, with S()-curves close to the rhodopsin-absorption functions, were found with _max at 332 nm (UV), 445 nm (blue) and 515 nm (green), respectively.Blue receptors were only recorded in the anatomically specialized dorsal rim area (DRA), and UV and green receptors in the dorsal region of the pigmented part of the eye, whereby green receptors were only found in the ventral eye. On the basis of these results, model calculations are presented for di- and trichromatic colour vision in the cricket.The fluorescence markings revealed green receptors whose axons project with short visual fibres to the lamina, and a UV receptor with a long visual fibre which projects through the lamina to the medulla. The blue receptors send their axons either to the lamina and medulla (long visual fibres) or only to the lamina (short visual fibres).The temporal dynamics of the three receptor types were examined. The blue receptors lack a phasic component of the receptor potential, and the time from stimulus on-set to peak potential is strongly increased compared to the UV and green receptors. Light adaptation reduces the latency to less than half of the dark adapted state.Spectral adaptation experiments revealed an unidirectional coupling between UV and green receptors, and it was found that polarization sensitivity (PS) in blue cells was much higher (PS= 6.5±1.5) than that of UV (PS=1.76±0.05) and green (2.26±0.57) receptors. The functional aspects of the three receptor types are discussed with respect to the presented physiological and morphological data.Abbreviations DA dorsal area - DRA dorsal rim area - PS polarization sensitivity     

The Molecular Evolution of Visual Pigments of Freshwater Crayfishes (Decapoda: Cambaridae)

This study examines the diverse maximum wavelength absorption (λ_max) found in crayfishes (Decapoda: Cambaridae and Parastacidae) and the associated genetic variation in their opsin locus. We measured the wavelength absorption in the photoreceptors of six species that inhabit environments of different light intensities (i.e., burrows, streams, standing waters, and subterranean waters). Our results indicate that there is relatively little variation in λ_max (522–530 nm) among species from different genera and families. The existing variation did not correlate with the habitat differences of the crayfishes studied. We simultaneously sequenced the rhodopsin gene to identify the amino acid replacements that affect shifts in maximum wavelength absorption. We then related these to changes that correlated with shifts in λ_max by reconstructing ancestral character states using a maximum-likelihood approach. Using amino acid sequences obtained from five species (all were 301 amino acids in length), we identified a number of candidates for producing shifts of 4 to 8 nm in λ_max. These amino acid replacements occurred in similar regions to those involved in spectral shifts in vertebrates. Received: 12 March 1997 / Accepted: 3 June 1997     

Ultraviolet photopigment sensitivity and ocular media transmittance in gulls,with an evolutionary perspective

Gulls (Laridae excluding Sternidae) appear to be the only shorebirds (Charadriiformes) that have a short wavelength sensitive type 1 (SWS1) cone pigment opsin tuned to ultraviolet (UV) instead of violet. However, the apparent UV-sensitivity has only been inferred indirectly, via the interpretation that the presence of cysteine at the key amino acid position 90 in the SWS1 opsin confers UV sensitivity. Unless the cornea and the lens efficiently transmit UV to the retina, gulls might in effect be similar to violet-sensitive birds in spectral sensitivity even if they have an ultraviolet sensitive (UVS) SWS1 visual pigment. We report that the spectral transmission of the cornea and lens of great black-backed Larus marinus and herring gulls L. argentatus allow UV-sensitivity, having a λ_T0.5 value, 344 nm, similar to the ocular media of UV sensitive birds. By molecular sequencing of the second α-helical transmembrane region of the SWS1 opsin gene we could also infer that 15 herring gulls and 16 yellow-legged gulls L. michahellis, all base-pair identical, are genetically UV-sensitive.     

Photoreceptor types and their relation to the spectral and polarization sensitivities of clupeid fishes

We used compound action potential recordings from the optic nerve of anesthetized live fish to study the spectral and polarization sensitivities of the northern anchovy and the Pacific herring. The photoreceptor structure and cone mosaic type of the (illuminated) central retina was studied by microscopy. Both species showed a single peak spectral photopic sensitivity function with λ_max= 500 nm for the northern anchovy and λ_max= 520 nm for the herring. However, only the northern anchovy exhibited polarization sensitivity; the response was 180° periodic with maximum sensitivity to horizontal polarization. Similar to the bay anchovy (Fineran and Nicol 1978), the central retina of the northern anchovy showed bifid cone units with cone lamellae parallel to the cones' lengths. The herring, on the other hand, had twin cones arranged in rows with the same orientation and tangentially arranged lamellae. Our results support the hypothesis that bifid cone units act as orthogonal dichroic filters rendering anchovies polarization sensitive. The lack of polarization sensitivity in the herring suggests that twin cones may not be used in polarization sensitivity or that one orientation of polarization receptors is insufficient for the animal to detect polarization direction. Accepted: 8 December 1997     

Visual pigments, oil droplets, ocular media and cone photoreceptor distribution in two species of passerine bird: the blue tit (Parus caeruleus L.) and the blackbird (Turdus merula L.)

The spectral absorption characteristics of the retinal photoreceptors of the blue tit (Parus caeruleus) and blackbird (Turdus merula) were investigated using microspectrophotometry. The retinae of both species contained rods, double cones and four spectrally distinct types of single cone. Whilst the visual pigments and cone oil droplets in the other receptor types are very similar in both species, the wavelength of maximum sensitivity (λ_max) of long-wavelength-sensitive single and double cone visual pigment occurs at a shorter wavelength (557 nm) in the blackbird than in the blue tit (563 nm). Oil droplets located in the long-wavelength-sensitivesingle cones of both species cut off wavelengths below 570–573 nm, theoretically shifting cone peak spectral sensitivity some 40 nm towards the long-wavelength end of the spectrum. This raises the possibility that the precise λ_max of the long-wavelength-sensitive visual pigment is optimised for the visual function of the double cones. The distribution of cone photoreceptors across the retina, determined using conventional light and fluorescence microscopy, also varies between the two species and may reflect differences in their visual ecology. Accepted: 8 January 2000     

Spectral sensitivity of the green photoreceptor of winged pea aphids

Intracellular recordings are obtained from photoreceptors in the retina of winged (alate) pea aphids Acyrthosiphon pisum (Harris). The responses to monochromatic light, applied in 10‐nm steps over the range 320–650 nm, reveal that all recordings are from green receptors and the spectral sensitivity function of these photoreceptors peaks at 518 nm. A comparison between the spectral sensitivity of the green receptors and extracellular electroretinogram recordings suggests that additional sensitivity to the short‐wavelength light (ultraviolet and/or blue) is also likely to be present in the compound eye of pea aphids. An analysis of the pea aphid genome, comparing its translated nucleotide sequences with the those of the opsin genes of other insect species, supports this electrophysiological finding, although it could not be established whether A. pisum, in addition to the green receptor, has both blue and ultraviolet receptors in the compound eye. The implications of these results for the visual ecology of herbivorous insects are discussed.     

Three spectrally distinct photoreceptors in diurnal and nocturnal Australian ants

Ants are thought to be special among Hymenopterans in having only dichromatic colour vision based on two spectrally distinct photoreceptors. Many ants are highly visual animals, however, and use vision extensively for navigation. We show here that two congeneric day- and night-active Australian ants have three spectrally distinct photoreceptor types, potentially supporting trichromatic colour vision. Electroretinogram recordings show the presence of three spectral sensitivities with peaks (λ_max) at 370, 450 and 550 nm in the night-active Myrmecia vindex and peaks at 370, 470 and 510 nm in the day-active Myrmecia croslandi. Intracellular electrophysiology on individual photoreceptors confirmed that the night-active M. vindex has three spectral sensitivities with peaks (λ_max) at 370, 430 and 550 nm. A large number of the intracellular recordings in the night-active M. vindex show unusually broad-band spectral sensitivities, suggesting that photoreceptors may be coupled. Spectral measurements at different temporal frequencies revealed that the ultraviolet receptors are comparatively slow. We discuss the adaptive significance and the probability of trichromacy in Myrmecia ants in the context of dim light vision and visual navigation.     

Photoreceptor spectral sensitivity in island and mainland populations of the bumblebee, <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Most species of flower-visiting Hymenoptera are trichromatic, with photoreceptor spectral sensitivity peaks in the UV, blue and green regions of the spectrum. Red flowers, therefore, should be relatively difficult to detect for such insects. Nevertheless, in population biological studies in the bumblebee, Bombus terrestris, the Sardinian island population (B. t. sassaricus) displayed significantly higher responses to red artificial flowers (in tests of innate colour choice and detectability) than several mainland populations of the same species (Chittka et al. in Cognitive ecology of pollination, pp 106–126, 2001; Popul Ecol 46:243–251, 2004). Since there is relatively little physiological data on population differences in sensory systems, we used intracellular recording to compare photoreceptor spectral sensitivity in B. t. sassaricus and the southern European and Mediterranean population, B. t. dalmatinus. The results show both populations to be UV–blue–green trichromats, but with a small but significant increase in long-wave sensitivity in island bees. Spectral peaks were estimated at 348, 435 and 533 nm (B. t. dalmatinus) and 347, 436 and 538 nm (B. t. sassaricus) for UV, blue and green receptors, respectively. There were no significant differences in UV and blue receptor sensitivities. We found no photoreceptors maximally sensitive to red spectral light in the Sardinian population and model calculations indicate that the behavioural population differences in colour responses cannot be directly explained by receptor population differences. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Sequence and localization of an ultraviolet (sws1) opsin in the retina of the Japanese sardine Sardinops melanostictus (Teleostei: Clupeiformes)

A full‐length complementary (c)DNA encoding ultraviolet (UV)‐sensitive opsin (sws1) was isolated from the retina of the Japanese sardine Sardinops melanostictus. The sws1 phylogenetic tree showed a sister group relationship with the Cypriniformes, following the ray‐finned fish phylogeny. By expressing reconstituted opsin in vitro, it was determined that the maximum absorbance spectrum (λ_max) of sws1 is around 382 nm, being intermediate in position between two subtypes of sws1 pigment that are UV sensitive (λ_max = 355–380 nm) and violet sensitive (λ_max = 388–455 nm), which have been reported to date. The ocular media transmitted >20% transmittance of light in the range of 360–600 nm. In situ hybridization analyses revealed that sws1 messenger (m)RNA is localized in a central single cone surrounded by four double cones in a square mosaic. The square mosaic occupies the ventro‐temporal quadrant of the retina and the in situ hybridization signals were dominant in this area suggesting that the fish may use UV vision when looking upward. Based on these results, considerable significances of potential UV sensitivity, in relation to characteristic habits of S. melanostictus, are discussed.     

Aromatic amino acid oxidation products as antioxidants

The accumulation of UV photolysis products of amino acids tyrosine and tryptophan, which possess antioxidant activity, has been studied by the method of luminol-dependent chemiluminescence. The amount of antioxidant products was judged by the value of the total antioxidant potential of a UV-irradiated solution, the measure of which was the distance between the peaks of the chemiluminescence curve in the system 2,2′-azo-bis(2-amidinopropane) hydrochloride + luminol with a UV-irradiated and an unirradiated sample (induction period, τ _i ). Simultaneously, the absorption and fluorescence spectra of unirradiared and UV-irradiated amino acid solutions were recorded. It was shown that exposure of a tryptophan solution to radiation led to accumulation of a fluorescent product N-formyl kynurenine (λ_em = 325 nm, λ_max = 440 nm), and the curve of its accumulation was similar to the growth of antioxidant potential. When a tyrosine solution was irradiated, the main fluorescent product was dityrosine (λ_em = 310 nm, λ_max = 415 nm). Nevertheless, the dose dependences of the formation of dityrosine and the total antioxidant potential were completely different. It was found that another product of tyrosine UV photolysis, dihydroxyphenylalanine, possessed pronounced antioxidant activity. It was concluded that the main antioxidant produced under UV irradiation of tryptophan is formyl kynurenine, and under irradiation of tyrosine it is dihydroxyphenylalanine.     

Temporal shifts in visual pigment absorbance in the retina of Pacific salmon

The visual pigments and photoreceptor types in the retinas of three species of Pacific salmon (coho, chum, and chinook) were examined using microspectrophotometry and histological sections for light microscopy. All three species had four cone visual pigments with maximum absorbance in the UV (_max: 357–382 nm), blue (_max: 431–446 nm), green (_max: 490–553 nm) and red (_max: 548–607 nm) parts of the spectrum, and a rod visual pigment with _max: 504–531 nm. The youngest fish (yolk-sac alevins) did not have blue visual pigment, but only UV pigment in the single cones. Older juveniles (smolts) had predominantly single cones with blue visual pigment. Coho and chinook smolts (>1 year old) switched from a vitamin A₁- to a vitamin A₂-dominated retina during the spring, while the retina of chum smolts and that of the younger alevin-to-parr coho did not. Adult spawners caught during the Fall had vitamin A₂-dominated retinas. The central retina of all species had three types of double cones (large, medium and small). The small double cones were situated toward the ventral retina and had lower red visual pigment _max than that of medium and large double cones, which were found more dorsally. Temperature affected visual pigment _max during smoltification.     

Estimated absorbance spectra of the visual pigments of the North Atlantic right whale (Eubalaena glacialis)

To assess the spectral sensitivities of the retinal visual pigments from the North Atlantic right whale (Eubalaena glacialis), we have cloned and sequenced two exons from the rod opsin gene and two exons from the middle‐wavelength sensitive (MWS) cone opsin gene in order to determine the amino acids at positions known to be key regulators of the spectral location of the absorbance maximum (λ_max). Based on previous mutagenesis models we estimate that the right whale possesses a rod visual pigment with a λ_max of 499 nm and a MWS cone visual pigment with a λ_max of 524 nm. Although the MWS cone visual pigment from the right whale is blue‐shifted in its spectral sensitivity like those from odontocetes, the spectral sensitivity of the right whale rod visual pigment is similar to those from terrestrial mammals.     

The Spectral Sensitivities of Single Cells in the Median Ocellus of Limulus

The spectral sensitivities of single Limulus median ocellus photoreceptors have been determined from records of receptor potentials obtained using intracellular microelectrodes. One class of receptors, called UV cells (ultraviolet cells), depolarizes to near-UV light and is maximally sensitive at 360 nm; a Dartnall template fits the spectral sensitivity curve. A second class of receptors, called visible cells, depolarizes to visible light; the spectral sensitivity curve is fit by a Dartnall template with λ_max at 530 nm. Dark-adapted UV cells are about 2 log units more sensitive than dark-adapted visible cells. UV cells respond with a small hyperpolarization to visible light and the spectral sensitivity curve for this hyperpolarization peaks at 525–550 nm. Visible cells respond with a small hyperpolarization to UV light, and the spectral sensitivity curve for this response peaks at 350–375 nm. Rarely, a double-peaked (360 and 530 nm) spectral sensitivity curve is obtained; two photopigments are involved, as revealed by chromatic adaptation experiments. Thus there may be a small third class of receptor cells containing two photopigments.     

Visual pigments and spectral sensitivity of the diurnal gecko Gonatodes albogularis

The visual pigments and oil droplets in the retina of the diurnal gecko Gonatodes albogularis were examined microspectrophotometrically, and the spectral sensitivity under various adapting conditions was recorded using electrophysiological responses. Three classes of visual pigments were identified, with _max at about 542, 475, and 362 nm. Spectral sensitivity functions revealed a broad range of sensitivity, with a peak at approximately 530–540 nm. The cornea and oil droplets were found to be transparent across a range from 350–700 nm, but the lens absorbed short wavelength light below 450 nm. Despite the filtering effect of the lens, a secondary peak in spectral sensitivity to ultraviolet wavelengths was found. These results suggest that G. albogularis does possess the visual mechanisms for discrimination of the color pattern of conspecifics based on either hue or brightness. These findings are discussed in terms of the variation in coloration and social behavior of Gonatodes.Abbreviations ERG electroretinogram - MSP microspectrophotometry - UV ultraviolet - _max wavelength of maximum absorbance     

Unconventional ultraviolet sensitivity spectra of Ascalaphus (Insecta,Neuroptera)

The spectral sensitivity of 21 eye preparations of Ascalaphus (Libelluloides) macaronius (Insecta, Neuroptera) has been re-measured using an up-to-date spectral scan method. 1. Dorso-frontal and ventro-lateral eyes have different spectral characteristics with peaks of sensitivity at 329 ± 8 nm (n = 15) and 343 ± 4 nm (n = 5) (P = 0.002), respectively. 2. The absorbance of the visual pigment layer, K, determined from the shape of the spectral sensitivity curves is 1.3 ± 1.8(n = 15) for dorso-frontal eyes and – 1.0 ± 0.3(n = 5) for ventrolateral eyes, thus implying higher selfscreening in the dorso-frontal eyes and narrowing of the spectral sensitivity curves as regards to a template visual pigment in ventro-lateral eyes. 3. Plotting K versus spectral sensitivity peak wavelength _max revealed an inverse correlation between these variables with K = 42.5 – 0.126 _max at r = 0.88(n = 19). 4. Extracts of ommochromes and carotenoids (Figs. 4 to 6) do not allow to account for the above diversity of optical properties of the Ascalaphus eye (Fig. 7).Abbreviations SSC spectral sensitivity curve - DF dorso-frontal eye - UV ultraviolet - VL ventro-lateral eye     

Photoreceptors and photopigments in a subterranean rodent,the pocket gopher (<Emphasis Type="Italic">Thomomys bottae</Emphasis>)

Pocket gophers (Thomomys bottae) are rodents that spend much of their lives in near-lightless subterranean burrows. The visual adaptations associated with this extreme environment were investigated by making anatomical observations of retinal organization and by recording retinal responses to photic stimulation. The size of the eye is within the normal range for rodents, the lens transmits light well down into the ultraviolet, and the retina conforms to the normal mammalian plan. Electroretinogram recording revealed the presence of three types of photopigments, a rod pigment with a spectral peak of about 495 nm and two types of cone pigment with respective peak values of about 367 nm (UV) and 505 nm (medium-wavelength sensitive). Both in terms of responsivity to lights varying in temporal frequency and in response recovery following intense light adaptation, the cone responses of the pocket gopher are similar to those of other rodents. Labeling experiments indicate an abundance of cones that reach densities in excess of 30,000 mm^–2. Cones containing UV opsin are found throughout the retina, but those containing medium-wavelength sensitive opsin are mostly restricted to the dorsal retina where coexpression of the two photopigments is apparently the rule. Rod densities are lower than those typical for nocturnal mammals.     

Microvillar orientation in the photoreceptors of the ant Cataglyphis bicolor

Polarization sensitivity in arthropod photoreceptors is crucially dependent on the arrangement of the microvilli within the rhabdom. Here, we present an electron-microscopical study in which the degree of microvillar alignment and changes in the cross-sectional areas of the rhabdoms along their length were studied in the compound eye of the desert ant, Cataglyphis bicolor. Serial cross-sections through the retina were taken and the orientation of the microvilli was determined in the photoreceptors of individually identified ommatidia. The reconstructions of microvillar alignment were made in the three anatomically and functionally distinct regions of the Cataglyphis compound eye: the dorsal rim area (DRA), the dorsal area (DA), and the ventral area (VA). The following morphological findings are consistent with polarization sensitivities measured previously by intracellular recordings. (1) The microvilli of the DRA photoreceptors are aligned in parallel along the entire length of the cell from the distal tip of the rhabdom down to its proximal end, near the basement membrane. The microvilli of the retinular cells R1 and R5 are always parallel to each other and perfectly perpendicular, with only minor deviation, to the microvillar orientation of the remaining receptor cells. (2) In the DA and VA regions of the eye, the microvillar tufts of the small receptors R1, R3, R5, R7, and R9 change their direction repetitively every 1-4 7m for up to 90°. In contrast, the large receptor cells R2, R4, R6, and R8 maintain their microvillar orientation rigidly. (3) In the DRA ommatidia, the cross-sectional areas of the rhabdomeres do not change along the length of the rhabdom, but substantial changes occur in the DA and VA ommatidia.