The visual pigments of four deep-sea crustacean species

Summary The visual pigments of four mesopelagic crustacean species were studied at sea by means of microspectrophotometry. The absorbance maxima obtained for the visual pigments and their metarhodopsins, respectively, were: 493 nm and 481 nm (Systellaspis debilis), 485 nm and 480 nm (Acanthephyra curtirostris), 491 nm and 482 nm (A. smithi), and 495 nm and 487 nm (Sergestes tenuiremis). The spectral characteristics of the rhodopsins and metarhodopsins permit high photosensitivity and facilitate photoregeneration in a nearly monochromatic environment. Photic regeneration of rhodopsins from the deep-sea environment was demonstrated, and data were obtained which are consistent with the occurrence of dark regeneration. Specific optical density of the observed visual pigments was calculated for two species.     

Interspecific variation in the visual pigments of deep-sea fishes

Summary Visual pigments in the rods of 38 species of deep-sea fish were examined by microspectrophotometry. 33 species were found to have a single rhodopsin with a wavelength of maximum absorbance ( _max) in the range 470–495 nm. Such visual pigments have absorbance maxima close to the wavelengths of maximum spectral transmission of oceanic water. 5 species, however, did not conform to this pattern and visual pigments were found with _max values ranging from 451 nm to 539 nm. In 4 of these species two visual pigments were found located in two types of rod. Some 2-pigment species which have unusual red sensitivity, also have red-emitting photophores. These species have both rhodopsin and porphyropsin pigments in their retinae, which was confirmed by HPLC, and the two pigments are apparently located in separate rods in the same retinal area. In deep-sea fishes the occurrence of unusual visual pigments seems to be correlated with aspects of the species' depth ranges. In addition to ecological influences we present evidence, in the form of _max spectral clustering, that indicates the degree of molecular constraint imposed on the evolution of visual pigments in the deep-sea.     

The cone photoreceptors and visual pigments of chameleons

Visual pigments, oil droplets and photoreceptor types in the retinas of four species of true chameleons have been examined by microspectrophotometry. The species occupy different photic environments: two species of Chamaeleo are from Madagascar and two species of Furcifer are from Africa and the Arabian Peninsula. In addition to double cones, four spectrally distinct classes of single cone were identified. No rod photoreceptors were observed. The visual pigments appear to be mixtures of rhodopsins and porphyropsins. Double cones contained a pale oil droplet in the principle member and both outer segments contained a long-wave-sensitive visual pigment with a spectral maximum between about 555 nm and 610 nm, depending on the rhodopsin/porphyropsin mixture. Long-wave-sensitive single cones contained a visual pigment spectrally identical to the double cones, but combined with a yellow oil droplet. The other three classes of single cone contained visual pigments with maxima at about 480–505, 440–450 and 375–385 nm, combined with yellow, clear and transparent oil droplets respectively. The latter two classes were sparsely distributed. The transmission of the lens and cornea of C. dilepis was measured and found to be transparent throughout the visible and near ultraviolet, with a cut off at about 350 nm.     

Photon Hunting in the Twilight Zone: Visual Features of Mesopelagic Bioluminescent Sharks

The mesopelagic zone is a visual scene continuum in which organisms have developed various strategies to optimize photon capture. Here, we used light microscopy, stereology-assisted retinal topographic mapping, spectrophotometry and microspectrophotometry to investigate the visual ecology of deep-sea bioluminescent sharks [four etmopterid species (Etmopterus lucifer, E. splendidus, E. spinax and Trigonognathus kabeyai) and one dalatiid species (Squaliolus aliae)]. We highlighted a novel structure, a translucent area present in the upper eye orbit of Etmopteridae, which might be part of a reference system for counterillumination adjustment or acts as a spectral filter for camouflage breaking, as well as several ocular specialisations such as aphakic gaps and semicircular tapeta previously unknown in elasmobranchs. All species showed pure rod hexagonal mosaics with a high topographic diversity. Retinal specialisations, formed by shallow cell density gradients, may aid in prey detection and reflect lifestyle differences; pelagic species display areae centrales while benthopelagic and benthic species display wide and narrow horizontal streaks, respectively. One species (E. lucifer) displays two areae within its horizontal streak that likely allows detection of conspecifics'' elongated bioluminescent flank markings. Ganglion cell topography reveals less variation with all species showing a temporal area for acute frontal binocular vision. This area is dorsally extended in T. kabeyai, allowing this species to adjust the strike of its peculiar jaws in the ventro-frontal visual field. Etmopterus lucifer showed an additional nasal area matching a high rod density area. Peak spectral sensitivities of the rod visual pigments (λ_max) fall within the range 484–491 nm, allowing these sharks to detect a high proportion of photons present in their habitat. Comparisons with previously published data reveal ocular differences between bioluminescent and non-bioluminescent deep-sea sharks. In particular, bioluminescent sharks possess higher rod densities, which might provide them with improved temporal resolution particularly useful for bioluminescent communication during social interactions.     

The visual pigments of a deep-sea myctophid fish Myctophum nitidulum Garman; an HPLC and spectroscopic description of a non-paired rhodopsin–porphyropsin system

Unusually for a deep-sea fish, the retina of the myctophid (lanternfish) Myctophum nitidulum was found to contain two visual pigments, shown by extract spectrophotometry to be maximally sensitive at 468 and 522 nm, respectively, giving this species one of the broadest spectral ranges of all deep-sea fishes. High performance liquid chromatography (HPLC) indicated that the retina contained both A₁- and A₂-based chromophores. Surprisingly, the maximum absorbance (λ_max) values of the two visual pigments were too far apart to form a rhodopsin–porphyropsin 'pigment pair', suggesting they were based on distinct opsins each linked to a different chromophore. This might be an adaptation to the detection of both long-wave bioluminescence and residual shorter-wave surface illumination, and could be related to this animal's tendency to migrate towards surface waters at night.     

Rod opsin sequence in the John Dory: further evidence for the spectral tuning of rhodopsin

The opsin from the John Dory Zeus faber rod visual pigment (maximum spectral sensitivity =492 nm) was cloned and sequenced. Comparison of the John Dory rod opsin sequence with those from other fish with blue-shifted scotopic spectral sensitivity provides further evidence for the spectral tuning of this group of visual pigments.     

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.     

Spectral tuning of avian violet- and ultraviolet-sensitive visual pigments

The violet- and ultraviolet-sensitive visual pigments of birds belong to the same class of pigments as the violet-sensitive (so-called blue) pigments of mammals. However, unlike the pigments from mammals and other vertebrate taxa which, depending on species, have lambda(max) values of either around 430 nm or around 370 nm, avian pigments are found with lambda(max) values spread across this range. In this paper, we present the sequences of two pigments isolated from Humbolt penguin and pigeon with intermediate lambda(max) values of 403 and 409 nm, respectively. By comparing the amino acid sequences of these pigments with the true UV pigments of budgerigar and canary and with chicken violet with a lambda(max) value of 420 nm, we have been able to identify five amino acid sites that show a pattern of substitution between species that is consistent with differences in lambda(max). Each of these substitutions has been introduced into budgerigar cDNA and expressed in vitro in COS-7 cells. Only three resulted in spectral shifts in the regenerated pigment; two had relatively small effects and may account for the spectral shifts between penguin, pigeon, and chicken whereas one, the replacement of Ser by Cys at site 90 in the UV pigments, produced a 35 nm shortwave shift that could account for the spectral shift from 403 nm in penguin to around 370 nm in budgerigar and canary.     

Vision in elasmobranchs and their relatives: 21st century advances

This review identifies a number of exciting new developments in the understanding of vision in cartilaginous fishes that have been made since the turn of the century. These include the results of studies on various aspects of the visual system including eye size, visual fields, eye design and the optical system, retinal topography and spatial resolving power, visual pigments, spectral sensitivity and the potential for colour vision. A number of these studies have covered a broad range of species, thereby providing valuable information on how the visual systems of these fishes are adapted to different environmental conditions. For example, oceanic and deep-sea sharks have the largest eyes amongst elasmobranchs and presumably rely more heavily on vision than coastal and benthic species, while interspecific variation in the ratio of rod and cone photoreceptors, the topographic distribution of the photoreceptors and retinal ganglion cells in the retina and the spatial resolving power of the eye all appear to be closely related to differences in habitat and lifestyle. Multiple, spectrally distinct cone photoreceptor visual pigments have been found in some batoid species, raising the possibility that at least some elasmobranchs are capable of seeing colour, and there is some evidence that multiple cone visual pigments may also be present in holocephalans. In contrast, sharks appear to have only one cone visual pigment. There is evidence that ontogenetic changes in the visual system, such as changes in the spectral transmission properties of the lens, lens shape, focal ratio, visual pigments and spatial resolving power, allow elasmobranchs to adapt to environmental changes imposed by habitat shifts and niche expansion. There are, however, many aspects of vision in these fishes that are not well understood, particularly in the holocephalans. Therefore, this review also serves to highlight and stimulate new research in areas that still require significant attention.     

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.     

On the visual pigments of deep-sea fish

The retinal visual pigments of 52 species of deep-sea fish were measured by partial bleaching of detergent extracts. The retinae of 45 species contained only a single rhodopsin with maximum absorbance (λ_max) at a wavelength between 474 and 490 nm, matching both the region of highest intensity downwelling sunlight and the maximum emission of most deep-sea bioluminescence. Seven species were shown to have more than one visual pigment within their retinae and these had λ_max values that generally fell outside the usual range. One of these, Bonapartia pedaliota , was particularly interesting as, unlike most such multipigment species, it had one rhodopsin and one porphyropsin pigment, apparently based on different opsins. The relative proportions of the visual pigments in the seven multipigment species are presented.     

Divergent mechanisms for the tuning of shortwave sensitive visual pigments in vertebrates.

Of the four classes of vertebrate cone visual pigments, the shortwave-sensitive SWS1 class shows the shortest lambda(max) values with peaks in different species in either the violet (390-435 nm) or ultraviolet (around 365 nm) regions of the spectrum. Phylogenetic evidence indicates that the ancestral pigment was probably UV-sensitive (UVS) and that the shifts between violet and UV have occurred many times during evolution. This is supported by the different mechanisms for these shifts in different species. All visual pigments possess a chromophore linked via a Schiff base to a Lys residue in opsin protein. In violet-sensitive (VS) pigments, the Schiff base is protonated whereas in UVS pigments, it is almost certainly unprotonated. The generation of VS from ancestral UVS pigments most likely involved amino acid substitutions in the opsin protein that serve to stabilise protonation. The key residues in the opsin protein for this are at sites 86 and 90 that are adjacent to the Schiff base and the counterion at Glu113. In this review, the different molecular mechanisms for the UV or violet shifts are presented and discussed in the context of the structural model of bovine rhodopsin.     

Visual predation during springtime foraging of the North Atlantic right whale (Eubalaena glacialis)

To assess the role that vision plays in the ability of the North Atlantic right whale (Eubalaena glacialis) to detect its primary prey species, the calanoid copepod Calanus finmarchicus, we have compared the absorbance spectrum of the E. glacialis rod visual pigment, the transmittance spectra of C. finmarchicus carotenoid pigments, as well as the downwelling irradiance and horizontal radiance spectra collected during springtime at three locations in the western Gulf of Maine. The E. glacialis rod visual pigment absorbs light maximally at 493 nm, while microspectrophotometric measurements of the C. finmarchicus carotenoid pigments reveal transmission spectra with minima matching very well with the E. glacialis rod visual pigment absorbance spectra maximum. Springtime spectral downwelling irradiance and horizontal radiance values from the surface waters of Cape Cod Bay and at all depths in Great South Channel overlap the E. glacialis rod absorbance spectrum, allowing C. finmarchicus to appear as a high‐contrast dark silhouette against a bright background spacelight, thus facilitating visually guided contrast foraging. In contrast, spectral downwelling irradiance and horizontal radiance at depth in Cape Cod Bay, and all depths in Wilkinson Basin, do not overlap the E. glacialis rod absorbance spectrum, providing little if any useful light for contrast vision.     

Switch in rod opsin gene expression in the European eel,Anguilla anguilla (L.).

The rod photoreceptors of the European eel, Anguilla anguilla (L.), alter their wavelength of maximum sensitivity (lambda max) from c.a. 523 nm to c.a. 482 nm at maturation, a switch involving the synthesis of a new visual pigment protein (opsin) that is inserted into the outer segments of existing rods. We artificially induced the switch in rod opsin production by the administration of hormones, and monitored the switch at the level of mRNA accumulation using radiolabelled oligonuleotides that hybridized differently to the two forms of eel rod opsin. The production of the deep-sea form of rod opsin was detected 6 h after the first hormone injection, and the switch in rod opsin expression was complete within four weeks, at which time only the mRNA for the deep-sea opsin was detectable in the retinal cells. It is suggested that this system could be used as a tractable model for studying the regulatory control of opsin gene expression.     

Long-wave sensitivity in deep-sea stomiid dragonfish with far-red bioluminescence: evidence for a dietary origin of the chlorophyll-derived retinal photosensitizer of Malacosteus niger

Both residual downwelling sunlight and bioluminescence, which are the two main sources of illumination available in the deep sea, have limited wavebands concentrated around 450-500 nm. Consequently, the wavelengths of maximum absorption (lambdamax) of the vast majority of deep-sea fish visual pigments also cluster in this part of the spectrum. Three genera of deep-sea loose-jawed dragonfish (Aristostomias, Pachystomias and Malacosteus), however, in addition to the blue bioluminescence typical of most deep-sea animals, also produce far-red light (maximum emission >700 nm) from suborbital photophores. All three genera are sensitive in this part of the spectrum, to which all other animals of the deep sea are blind, potentially affording them a private waveband for illuminating prey and for interspecific communication that is immune from detection by predators and prey. Aristostomias and Pachystomias enhance their long-wave visual sensitivity by the possession of at least three visual pigments that are long-wave shifted (lambdamax values ca. 515, 550 and 590 nm) compared with those of other deep-sea fishes. Malacosteus, on the other hand, although it does possess two of these red-shifted pigments (lambdamax values ca. 520 and 540 nm), lacks the most long-wave-sensitive pigments found in the other two genera. However, it further enhances its long-wave sensitivity with a chlorophyll-derived photosensitizer within its outer segments. The fluorescence emission and excitation spectra of this pigment are very similar to spectra obtained from mesopelagic copepods, which are an important component of diet of Malacosteus, suggesting a dietary origin for this pigment.     

A novel spectral tuning in the short wavelength-sensitive (SWS1 and SWS2) pigments of bluefin killifish (Lucania goodei)

The molecular bases of spectral tuning in the UV-, violet-, and blue-sensitive pigments are not well understood. Using the in vitro assay, here we show that the SWS1, SWS2-A, and SWS2-B pigments of bluefin killifish (Lucania goodei) have the wavelengths of maximal absorption (lambda(max)'s) of 354, 448, and 397 nm, respectively. The spectral difference between the SWS2-A and SWS2-B pigments is largest among those of all currently known pairs of SWS2 pigments within a species. The SWS1 pigment contains no amino acid replacement at the currently known 25 critical sites and seems to have inherited its UV-sensitivity directly from the vertebrate ancestor. Mutagenesis analyses show that the amino acid differences at sites 44, 46, 94, 97, 109, 116, 118, 265, and 292 of the SWS2-A and SWS2-B pigments explain 80% of their spectral difference. Moreover, the larger the individual effects of amino acid changes on the lambda(max)-shift are, the larger the synergistic effects tend to be generated, revealing a novel mechanism of spectral tuning of visual pigments.     

Optimization, constraint, and history in the evolution of eyes

Several features of the evolution of eyes and photoreceptors are examined in an effort to explore the relative roles of adaptation and historical and developmental constraints. Optical design shows clear evidence of adaptation, which in some respects approaches optima predictable from physics. The primate fovea, on the other hand, illustrates how adaptation can be channeled by developmental heritage. The primary structures of opsins reveal multiple evolutionary lineages within both Drosophila and humans. The pigments of vertebrae rods comprise a subset of opsins whose evolutionary relationships map onto the phylogeny of the parent species. The evolutionary reasons for why most rod pigments absorb maximally at 500 +/- 10 nm are obscure, as there is no convincing explanation based on adaptation alone. Rods are appropriately distinguished from cones on the basis of which opsin gene is expressed. This criterion is likely to be in conflict with other definitions in phyletic lines (e.g., geckos, snakes) that have long diurnal or nocturnal histories accompanied by loss of one or more opsin genes, followed by a secondary adaptation to life in a different photic environment. Color vision--a generalizable perception associated with the spectral composition of light--is usefully distinguished from wavelength-specific behaviors. The latter are also based on multiple visual pigments and more than one spectral class of receptors but cannot be altered by learning. The distinction is particularly forceful in bees, which exhibit both kinds of behavior. The evolution of primate color vision has been shaped by historical factors involving an extensive period of early mammalian nocturnality. Birds, by contrast, have more elaborate cones and a richer set of visual pigments. Avian color space can be represented in a tetrahedron.     

Cone opsin genes of african cichlid fishes: tuning spectral sensitivity by differential gene expression

Spectral tuning of visual pigments is typically accomplished through changes in opsin amino acid sequence. Within a given opsin class, changes at a few key sites control wavelength specificity. To investigate known differences in the visual pigment spectral sensitivity of the Lake Malawi cichlids, Metriaclima zebra (368, 488, and 533 nm) and Dimidiochromis compressiceps (447, 536, and 569 nm), we sequenced cone opsin genes from these species as well as Labeotropheus fuelleborni and Oreochromis niloticus. These cichlids have five distinct classes of cone opsin genes, including two unique SWS-2 genes. Comparisons of the inferred amino acid sequences from the five cone opsin genes of M. zebra, D. compressiceps, and L. fuelleborni show the sequences to be nearly identical. Therefore, evolution of key opsin sites cannot explain the differences in visual pigment sensitivities. Real-time PCR demonstrates that different cichlid species express different subsets of the available cone opsin genes. Metriaclima zebra and L. fuelleborni express a complement of genes which give them UV-shifted visual pigments, while D. compressiceps expresses a different set to produce a red-shifted visual system. Thus, variations in cichlid spectral sensitivity have arisen through evolution of gene regulation, rather than through changes in opsin amino acid sequence.     

Visual pigments in the individual rods of deep-sea fishes

Summary The visual pigments in the rods of 15 species of deep-sea fish were examined by microspectrophotometry. In 13 species a single visual pigment was found. The max of these pigments, which ranged from 475 nm to 488 nm, suggest they give the fish maximum sensitivity to the ambient light in the deep, blue ocean waters where they live. In two species two visual pigments were found in separate rods.Bathylagus bericoides had rhodopsins of max 466 nm and 500 nm andMalacocephalus laevis had two rhodopsins of max 478 nm and 485 nm. It is noted that the species with two visual pigments tend to be dark in colour and live in deeper, darker, water.