Ultraviolet vision in anemonefish improves colour discrimination.

In many animals, ultraviolet (UV) vision guides navigation, foraging, and communication, but few studies have addressed the contribution of UV signals to colour vision, or measured UV discrimination thresholds using behavioural experiments. Here, we tested UV colour vision in an anemonefish (Amphiprion ocellaris) using a five-channel (RGB-V-UV) LED display. We first determined that the maximal sensitivity of the A. ocellaris UV cone was ∼386 nm using microspectrophotometry. Three additional cone spectral sensitivities had maxima at ∼497, 515 and ∼535 nm. We then behaviourally measured colour discrimination thresholds by training anemonefish to distinguish a coloured target pixel from grey distractor pixels of varying intensity. Thresholds were calculated for nine sets of colours with and without UV signals. Using a tetrachromatic vision model, we found that anemonefish were better (i.e. discrimination thresholds were lower) at discriminating colours when target pixels had higher UV chromatic contrast. These colours caused a greater stimulation of the UV cone relative to other cone types. These findings imply that a UV component of colour signals and cues improves their detectability, which likely increases the prominence of anemonefish body patterns for communication and the silhouette of zooplankton prey.

Multiple rod layers increase the speed and sensitivity of vision in nocturnal reef fishes.

Most vertebrates have one layer of the dim-light active rod photoreceptors. However, multiple rod layers, known as a multibank retina, can be found in over 100 species of fish, including several deep-sea species and one family of nocturnally active reef fish, the Holocentridae. Although seemingly associated with increased photon catch, the function of multibank retinas remained unknown. We used an integrative approach, combining histology, electrophysiology and amino acid sequence analysis, applied to three species of nocturnal reef fishes, two holocentrids with a multibank retina (Neoniphon sammara and Myripristis violacea) and an apogonid with a single rod bank (Ostorhinchus compressus), to determine the sensory advantage of multiple rod layers. Our results showed that fish with multibank retinas have both faster vision and enhanced responses to bright- and dim-light intensities. Faster vision was indicated by higher flicker fusion frequencies during temporal resolution electroretinography as well as faster retinal release rates estimated from their rhodopsin proteins. Enhanced sensitivity was demonstrated by broadened intensity-response curves derived from luminous sensitivity electroretinography. Overall, our findings provide the first functional evidence for enhanced dim-light sensitivity using a multibank retina while also suggesting novel roles for the adaptation in enhancing bright-light sensitivity and the speed of vision.

Comparative visual ecology of cephalopods from different habitats.

Previous investigations of vision and visual pigment evolution in aquatic predators have focused on fish and crustaceans, generally ignoring the cephalopods. Since the first cephalopod opsin was sequenced in late 1980s, we now have data on over 50 cephalopod opsins, prompting this functional and phylogenetic examination. Much of this data does not specifically examine the visual pigment spectral absorbance position (λmax) relative to environment or lifestyle, and cephalopod opsin functional adaptation and visual ecology remain largely unknown. Here we introduce a new protocol for photoreceptor microspectrophotometry (MSP) that overcomes the difficulty of bleaching the bistable visual pigment and that reveals eight coastal coleoid cephalopods to be monochromatic with λmax varying from 484 to 505 nm. A combination of current MSP results, the λmax values previously characterized using cephalopod retinal extracts (467-500 nm) and the corresponding opsin phylogenetic tree were used for systematic comparisons with an end goal of examining the adaptations of coleoid visual pigments to different light environments. Spectral tuning shifts are described in response to different modes of life and light conditions. A new spectral tuning model suggests that nine amino acid substitution sites may determine the direction and the magnitude of spectral shifts.

Ocean acidification slows retinal function in a damselfish through interference with GABAA receptors.

Vision is one of the most efficient senses used by animals to catch prey and avoid predators. Therefore, any deficiency in the visual system could have important consequences for individual performance. We examined the effect of CO2 levels projected to occur by the end of this century on retinal responses in a damselfish, by determining the threshold of its flicker electroretinogram (fERG). The maximal flicker frequency of the retina was reduced by continuous exposure to elevated CO2, potentially impairing the capacity of fish to react to fast events. This effect was rapidly counteracted by treatment with a GABA antagonist (gabazine), indicating that GABAA receptor function is disrupted by elevated CO2. In addition to demonstrating the effects of elevated CO2 on fast flicker fusion of marine fishes, our results show that the fish retina could be a model system to study the effects of high CO2 on neural processing.

The brain structure and the neural network features of the diurnal cuttlefish Sepia plangon.

Cuttlefish are known for their rapid changes of appearance enabling camouflage and con-specific communication for mating or agonistic display. However, interpretation of their sophisticated behaviors and responsible brain areas is based on the better-studied squid brain atlas. Here we present the first detailed description of the neuroanatomical features of a tropical and diurnal cuttlefish, Sepia plangon, coupled with observations on ontogenetic changes in its visual and learning centers using a suite of MRI-based techniques and histology. We then make comparisons to a loliginid squid, treating it as a 'baseline', and also to other cuttlefish species to help construct a connectivity map of the cuttlefish brain. Differences in brain anatomy and the previously unknown neural connections associated with camouflage, motor control and chemosensory function are described. These findings link brain heterogeneity to ecological niches and lifestyle, feeding hypotheses and evolutionary history, and provide a timely, new technology update to older literature.

Comparative brain structure and visual processing in octopus from different habitats.

Octopods are masters of camouflage and solve complex tasks, and their cognitive ability is said to approach that of some small mammals. Despite intense interest and some research progress, much of our knowledge of octopus neuroanatomy and its links to behavior and ecology comes from one coastal species, the European common octopus, Octopus vulgaris. Octopod species are found in habitats including complex coral reefs and the relatively featureless mid-water. There they encounter different selection pressures, may be nocturnal or diurnal, and are mostly solitary or partially social. How these different ecologies and behavioral differences influence the octopus central nervous system (CNS) remains largely unknown. Here we present a phylogenetically informed comparison between diurnal and nocturnal coastal and a deep-sea species using brain imaging techniques. This study shows that characteristic neuroanatomical changes are linked to their habits and habitats. Enlargement and division of the optic lobe as well as structural foldings and complexity in the underlying CNS are linked to behavioral adaptation (diurnal versus nocturnal; social versus solitary) and ecological niche (reef versus deep sea), but phylogeny may play a part also. The difference between solitary and social life is mirrored within the brain including the formation of multiple compartments (gyri) in the vertical lobe, which is likened to the vertebrate cortex. These findings continue the case for convergence between cephalopod and vertebrate brain structure and function. Notably, within the current push toward comparisons of cognitive abilities, often with unashamed anthropomorphism at their root, these findings provide a firm grounding from which to work.

Dynamic Courtship Signals and Mate Preferences in Sepia plangon.

Communication in cuttlefish includes rapid changes in skin coloration and texture, body posture and movements, and potentially polarized signals. The dynamic displays are fundamental for mate choice and agonistic behavior. We analyzed the reproductive behavior of the mourning cuttlefish Sepia plangon in the laboratory. Mate preference was analyzed via choice assays (n = 33) under three sex ratios, 1 male (M): 1 female (F), 2M:1F, and 1M:2F. We evaluated the effect of modifying polarized light from the arms stripes and ambient light with polarized and unpolarized barriers between the cuttlefish. Additionally, to assess whether a particular trait was a determinant for mating, we used 3D printed cuttlefish dummies. The dummies had different sets of visual signals: two sizes (60 or 90 mm mantle length), raised or dropped arms, high or low contrast body coloration, and polarized or unpolarized filters to simulate the arms stripes. Frequency and duration (s) of courtship displays, mating, and agonistic behaviors were analyzed with GLM and ANOVAs. The behaviors, body patterns, and their components were integrated into an ethogram to describe the reproductive behavior of S. plangon. We identified 18 body patterns, 57 body patterns components, and three reproductive behaviors (mating, courtship, and mate guarding). Only sex ratio had a significant effect on courtship frequency, and the male courtship success rate was 80%. Five small (ML < 80 mm) males showed the dual-lateral display to access mates while avoiding fights with large males; this behavior is characteristic of male "sneaker" cuttlefish. Winner males showed up to 17 body patterns and 33 components, whereas loser males only showed 12 patterns and 24 components. We identified 32 combinations of body patterns and components that tended to occur in a specific order and were relevant for mating success in males. Cuttlefish were visually aware of the 3D-printed dummies; however, they did not start mating or agonistic behavior toward the dummies. Our findings suggest that in S. plangon, the dynamic courtship displays with specific sequences of visual signals, and the sex ratio are critical for mate choice and mating success.

Toward an MRI-Based Mesoscale Connectome of the Squid Brain.

Using high-resolution diffusion magnetic resonance imaging (dMRI) and a suite of old and new staining techniques, the beginnings of a multi-scale connectome map of the squid brain is erected. The first of its kind for a cephalopod, this includes the confirmation of 281 known connections with the addition of 145 previously undescribed pathways. These and other features suggest a suite of functional attributes, including (1) retinotopic organization through the optic lobes and into other brain areas well beyond that previously recognized, (2) a level of complexity and sub-division in the basal lobe supporting ideas of convergence with the vertebrate basal ganglia, and (3) differential lobe-dependent growth rates that mirror complexity and transitions in ontogeny.

Acoustically evoked potentials in two cephalopods inferred using the auditory brainstem response (ABR) approach.

It is still a matter of debate whether cephalopods can detect sound frequencies above 400 Hz. So far there is no proof for the detection of underwater sound above 400 Hz via a physiological approach. The controversy of whether cephalopods have a sound detection ability above 400 Hz was tested using the auditory brainstem response (ABR) approach, which has been successfully applied in fish, crustaceans, amphibians, reptiles and birds. Using ABR we found that auditory evoked potentials can be obtained in the frequency range 400 to 1500 Hz (Sepiotheutis lessoniana) and 400 to 1000 Hz (Octopus vulgaris), respectively. The thresholds of S. lessoniana were generally lower than those of O. vulgaris.

A detailed investigation of the visual system and visual ecology of the Barrier Reef anemonefish, Amphiprion akindynos.

Vision plays a major role in the life of most teleosts, and is assumingly well adapted to each species ecology and behaviour. Using a multidisciplinary approach, we scrutinised several aspects of the visual system and ecology of the Great Barrier Reef anemonefish, Amphiprion akindynos, including its orange with white patterning, retinal anatomy and molecular biology, its symbiosis with anemones and sequential hermaphroditism. Amphiprion akindynos possesses spectrally distinct visual pigments and opsins: one rod opsin, RH1 (498 nm), and five cone opsins, SWS1 (370 nm), SWS2B (408 nm), RH2B (498 nm), RH2A (520 nm), and LWS (554 nm). Cones were arranged in a regular mosaic with each single cone surrounded by four double cones. Double cones mainly expressed RH2B (53%) in one member and RH2A (46%) in the other, matching the prevailing light. Single cones expressed SWS1 (89%), which may serve to detect zooplankton, conspecifics and the host anemone. Moreover, a segregated small fraction of single cones coexpressed SWS1 with SWS2B (11%). This novel visual specialisation falls within the region of highest acuity and is suggested to increase the chromatic contrast of Amphiprion akindynos colour patterns, which might improve detection of conspecifics.

Adaptive evolution of cone opsin genes in two colorful cyprinids, Opsariichthys pachycephalus and Candidia barbatus.

Opsariichthys pachycephalus and Candidia barbatus are two phylogenetically related freshwater cyprinids that both exhibit colorful, yet quite different nuptial coloration. This study was designed to test the hypothesis that differences in nuptial coloration between two species could reflect differences in color perception ability and the opsin genes that coded for it. Genes encoding the visual pigments of these two species were cloned and sequenced, lambda(max) of cone photoreceptors and the reflectance spectra of their body coloration were measured to test the hypothesis. The 14-nm spectral shift between green-light-sensitive photoreceptors of these two cyprinids is found to correlate well with differences in their reflective spectra. The spectral shift could result from differential expression of opsin genes and the interactive effects of the amino acid replacements in various minor sites. These results support our hypothesis that nuptial coloration is tied to color perception ability and opsin genes.

Morphological changes of the optic lobe from late embryonic to adult stages in oval squids Sepioteuthis lessoniana.

The optic lobe is the largest brain area within the central nervous system of cephalopods and it plays important roles in the processing of visual information, the regulation of body patterning, and locomotive behavior. The oval squid Sepioteuthis lessoniana has relatively large optic lobes that are responsible for visual communication via dynamic body patterning. It has been observed that the visual behaviors of oval squids change as the animals mature, yet little is known about how the structure of the optic lobes changes during development. The aim of the present study was to characterize the ontogenetic changes in neural organization of the optic lobes of S. lessoniana from late embryonic stage to adulthood. Magnetic resonance imaging and micro-CT scans were acquired to reconstruct the 3D-structure of the optic lobes and examine the external morphology at different developmental stages. In addition, optic lobe slices with nuclear staining were used to reveal changes in the internal morphology throughout development. As oval squids mature, the proportion of the brain making up the optic lobes increases continuously, and the optic lobes appear to have a prominent dent on the ventrolateral side. Inside the optic lobe, the cortex and the medulla expand steadily from the late embryonic stage to adulthood, but the cell islands in the tangential zone of the optic lobe decrease continuously in parallel. Interestingly, the size of the nuclei of cells within the medulla of the optic lobe increases throughout development. These findings suggest that the optic lobe undergoes continuous external morphological change and internal neural reorganization throughout the oval squid's development. These morphological changes in the optic lobe are likely to be responsible for changes in the visuomotor behavior of oval squids from hatching to adulthood.

Complex Visual Adaptations in Squid for Specific Tasks in Different Environments.

In common with their major competitors, the fish, squid are fast moving visual predators that live over a great range of depths in the ocean. Both squid and fish show a variety of adaptations with respect to optical properties, receptors and their underlying neural circuits, and these adaptations are often linked to the light conditions of their specific niche. In contrast to the extensive investigations of adaptive strategies in fish, vision in response to the varying quantity and quality of available light, our knowledge of visual adaptations in squid remains sparse. This study therefore undertook a comparative study of visual adaptations and capabilities in a number of squid species collected between 0 and 1,200 m. Histology, magnetic resonance imagery (MRI), and depth distributions were used to compare brains, eyes, and visual capabilities, revealing that the squid eye designs reflect the lifestyle and the versatility of neural architecture in its visual system. Tubular eyes and two types of regional retinal deformation were identified and these eye modifications are strongly associated with specific directional visual tasks. In addition, a combination of conventional and immuno-histology demonstrated a new form of a complex retina possessing two inner segment layers in two mid-water squid species which they rhythmically move across a broad range of depths (50-1,000 m). In contrast to their relatives with the regular single-layered inner segment retina live in the upper mesopelagic layer (50-400 m), the new form of retinal interneuronal layers suggests that the visual sensitivity of these two long distance vertical migrants may increase in response to dimmer environments.

Dynamic Skin Patterns in Cephalopods.

Cephalopods are unrivaled in the natural world in their ability to alter their visual appearance. These mollusks have evolved a complex system of dermal units under neural, hormonal, and muscular control to produce an astonishing variety of body patterns. With parallels to the pixels on a television screen, cephalopod chromatophores can be coordinated to produce dramatic, dynamic, and rhythmic displays, defined collectively here as "dynamic patterns." This study examines the nature, context, and potential functions of dynamic patterns across diverse cephalopod taxa. Examples are presented for 21 species, including 11 previously unreported in the scientific literature. These range from simple flashing or flickering patterns, to highly complex passing wave patterns involving multiple skin fields.

Range-finding in squid using retinal deformation and image blur.

Squid and other cephalopods catch prey with remarkable speed and precision [1]. Before the strike occurs, they encounter the difficult task of judging an object's distance and size in the contrast-poor world of the mid-water environment [1-4]. Here we describe a solution to this common problem underwater, where a large portion of a squid's dorso-temporal retina is intentionally blurred. This apparently counter-adaptive 'retinal bump' is combined with a vertical bobbing behavior that scans objects of interest from focused to defocused retinal regions. The image focus differential changes sharply at precisely the distance equivalent to tentacle length and enables the squid, Sepioteuthis lessoniana, to capture prey. This unique range-finding mechanism is an adaptation to hunting, defense, and object size identification in an environment where the depth cues found on land are less reliable.