Lydia M. Mäthger

Mechanisms and behavioural functions of structural coloration in cephalopods

Octopus, squid and cuttlefish are renowned for rapid adaptive coloration that is used for a wide range of communication and camouflage. Structural coloration plays a key role in augmenting the skin patterning that is produced largely by neurally controlled pigmented chromatophore organs. While most iridescence and white scattering is produced by passive reflectance or diffusion, some iridophores in squid are actively controlled via a unique cholinergic, non-synaptic neural system. We review the recent anatomical and experimental evidence regarding the mechanisms of reflection and diffusion of light by the different cell types (iridophores and leucophores) of various cephalopod species. The structures that are responsible for the optical effects of some iridophores and leucophores have recently been shown to be proteins. Optical interactions with the overlying pigmented chromatophores are complex, and the recent measurements are presented and synthesized. Polarized light reflected from iridophores can be passed through the chromatophores, thus enabling the use of a discrete communication channel, because cephalopods are especially sensitive to polarized light. We illustrate how structural coloration contributes to the overall appearance of the cephalopods during intra- and interspecific behavioural interactions including camouflage.

Cephalopod dynamic camouflage: bridging the continuum between background matching and disruptive coloration

Individual cuttlefish, octopus and squid have the versatile capability to use body patterns for background matching and disruptive coloration. We define—qualitatively and quantitatively—the chief characteristics of the three major body pattern types used for camouflage by cephalopods: uniform and mottle patterns for background matching, and disruptive patterns that primarily enhance disruptiveness but aid background matching as well. There is great variation within each of the three body pattern types, but by defining their chief characteristics we lay the groundwork to test camouflage concepts by correlating background statistics with those of the body pattern. We describe at least three ways in which background matching can be achieved in cephalopods. Disruptive patterns in cuttlefish possess all four of the basic components of ‘disruptiveness’, supporting Cotts hypotheses, and we provide field examples of disruptive coloration in which the body pattern contrast exceeds that of the immediate surrounds. Based upon laboratory testing as well as thousands of images of camouflaged cephalopods in the field (a sample is provided on a web archive), we note that size, contrast and edges of background objects are key visual cues that guide cephalopod camouflage patterning. Mottle and disruptive patterns are frequently mixed, suggesting that background matching and disruptive mechanisms are often used in the same pattern.

Rapid colour changes in multilayer reflecting stripes in the paradise whiptail, Pentapodus paradiseus

SUMMARY The Paradise whiptail (Pentapodus paradiseus) has distinct reflective stripes on its head and body. The reflective stripes contain a dense layer of physiologically active iridophores, which act as multilayer reflectors. The wavelengths reflected by these stripes can change from blue to red in 0.25 s. Transmission electron microscopy revealed that the iridophore cells contain plates that are, on average, 51.4 nm thick. This thickness produces a stack, which acts as an ideal quarter-wavelength multilayer reflector (equal optical thickness of plates and spaces) in the blue, but not the red, region of the spectrum. When skin preparations were placed into hyposmotic physiological saline, the peak wavelength of the reflected light shifted towards the longer (red) end of the visible spectrum. Hyperosmotic saline reversed this effect and shifted the peak wavelength towards shorter (blue/UV) wavelengths. Norepinephrine (100 μmol l-1) shifted the peak wavelength towards the longer end of the spectrum, while adenosine (100μ mol l-1) reversed the effects of norepinephrine. The results from this study show that the wavelength changes are elicited by a change of∼ 70 nm in the distance between adjacent plates in the iridophore cells.

Malleable skin coloration in cephalopods: selective reflectance, transmission and absorbance of light by chromatophores and iridophores

Nature’s best-known example of colorful, changeable, and diverse skin patterning is found in cephalopods. Color and pattern changes in squid skin are mediated by the action of thousands of pigmented chromatophore organs in combination with subjacent light-reflecting iridophore cells. Chromatophores (brown, red, yellow pigment) are innervated directly by the brain and can quickly expand and retract over underlying iridophore cells (red, orange, yellow, green, blue iridescence). Here, we present the first spectral account of the colors that are produced by the interaction between chromatophores and iridophores in squid (Loligo pealeii). Using a spectrometer, we have acquired highly focused reflectance measurements of chromatophores, iridophores, and the quality and quantity of light reflected when both interact. Results indicate that the light reflected from iridophores can be filtered by the chromatophores, enhancing their appearance. We have also measured polarization aspects of iridophores and chromatophores and show that, whereas structurally reflecting iridophores polarize light at certain angles, pigmentary chromatophores do not. We have further measured the reflectance change that iridophores undergo during physiological activity, from “off” to various degrees of “on”, revealing specifically the way that colors shift from the longer end (infra-red and red) to the shorter (blue) end of the spectrum. By demonstrating that three color classes of pigments, combined with a single type of reflective cell, produce colors that envelop the whole of the visible spectrum, this study provides an insight into the optical mechanisms employed by the elaborate skin of cephalopods to give the extreme diversity that enables their dynamic camouflage and signaling.

Changes in reflectin protein phosphorylation are associated with dynamic iridescence in squid

Many cephalopods exhibit remarkable dermal iridescence, a component of their complex, dynamic camouflage and communication. In the species Euprymna scolopes, the light-organ iridescence is static and is due to reflectin protein-based platelets assembled into lamellar thin-film reflectors called iridosomes, contained within iridescent cells called iridocytes. Squid in the family Loliginidae appear to be unique in which the dermis possesses a dynamic iridescent component with reflective, coloured structures that are assembled and disassembled under the control of the muscarinic cholinergic system and the associated neurotransmitter acetylcholine (ACh). Here we present the sequences and characterization of three new members of the reflectin family associated with the dynamically changeable iridescence in Loligo and not found in static Euprymna iridophores. In addition, we show that application of genistein, a protein tyrosine kinase inhibitor, suppresses ACh- and calcium-induced iridescence in Loligo. We further demonstrate that two of these novel reflectins are extensively phosphorylated in concert with the activation of iridescence by exogenous ACh. This phosphorylation and the correlated iridescence can be blocked with genistein. Our results suggest that tyrosine phosphorylation of reflectin proteins is involved in the regulation of dynamic iridescence in Loligo.

Anatomical basis for camouflaged polarized light communication in squid

Abstract Camouflage is a means to defeat visual detection by predators, whereas visual communication involves a signal that is conspicuous to a receiver (usually a conspecific). However, most intraspecific visual signals are also conspicuous to predators, so that signalling can lead to the serious consequence of predation. Could an animal achieve visual camouflage and simultaneously send a hidden visual message to a conspecific? Here, we present evidence that the polarized aspect of iridescent colour in squid skin is maintained after it passes through the overlying pigmented chromatophores, which produce the highly evolved—and dynamically changeable—camouflaged patterns in cephalopods. Since cephalopods are polarization sensitive, and can regulate polarization via skin iridescence, it is conceivable that they could send polarized signals to conspecifics while staying camouflaged to fish or mammalian predators, most of which are not polarization sensitive.

Cuttlefish use visual cues to control three-dimensional skin papillae for camouflage.

Cephalopods (octopus, squid and cuttlefish) are known for their camouflage. Cuttlefish Sepia officinalis use chromatophores and light reflectors for color change, and papillae to change three-dimensional physical skin texture. Papillae vary in size, shape and coloration; nine distinct sets of papillae are described here. The objective was to determine whether cuttlefish use visual or tactile cues to control papillae expression. Cuttlefish were placed on natural substrates to evoke the three major camouflage body patterns: Uniform/Stipple, Mottle and Disruptive. Three versions of each substrate were presented: the actual substrate, the actual substrate covered with glass (removes tactile information) and a laminated photograph of the substrate (removes tactile and three-dimensional information because depth-of-field information is unavailable). No differences in Small dorsal papillae or Major lateral mantle papillae expression were observed among the three versions of each substrate. Thus, visual (not tactile) cues drive the expression of papillae in S. officinalis. Two sets of papillae (Major lateral mantle papillae and Major lateral eye papillae) showed irregular responses; their control requires future investigation. Finally, more Small dorsal papillae were shown in Uniform/Stipple and Mottle patterns than in Disruptive patterns, which may provide clues regarding the visual mechanisms of background matching versus disruptive coloration.

Biological versus electronic adaptive coloration: how can one inform the other?

Adaptive reflective surfaces have been a challenge for both electronic paper (e-paper) and biological organisms. Multiple colours, contrast, polarization, reflectance, diffusivity and texture must all be controlled simultaneously without optical losses in order to fully replicate the appearance of natural surfaces and vividly communicate information. This review merges the frontiers of knowledge for both biological adaptive coloration, with a focus on cephalopods, and synthetic reflective e-paper within a consistent framework of scientific metrics. Currently, the highest performance approach for both nature and technology uses colourant transposition. Three outcomes are envisioned from this review: reflective display engineers may gain new insights from millions of years of natural selection and evolution; biologists will benefit from understanding the types of mechanisms, characterization and metrics used in synthetic reflective e-paper; all scientists will gain a clearer picture of the long-term prospects for capabilities such as adaptive concealment and signalling.

Evidence for distributed light sensing in the skin of cuttlefish, Sepia officinalis

We report that the skin of cuttlefish, Sepia officinalis, contains opsin transcripts suggesting a possible role of distributed light sensing for dynamic camouflage and signalling. The mRNA coding for opsin from various body regions was amplified and sequenced, and gene expression was detected in fin and ventral skin samples. The amino acid sequence of the opsin polypeptide that these transcripts would produce was identical in retina and fin tissue samples, but the ventral skin opsin transcripts differed by a single amino acid. The diverse camouflage and signalling body patterns of cephalopods are visually controlled, and these findings suggest a possible additional mechanism of light sensing and subsequent skin patterning. Cuttlefish, along with a number of other cephalopod species, have been shown to be colour-blind. Since the opsin in the fin is identical to that of the retina (λmax = 492 nm), and the ventral transcripts are also unlikely to be spectrally different, colour discrimination by the skin opsins is unlikely. However, spectral discrimination could be provided by involving other skin structures (chromatophores and iridophores), which produce changeable colours and patterns. This ‘distributed sensing’ could supplement the otherwise visually driven dynamic camouflage system by assisting with colour or brightness matching to adjacent substrates.

Color matching on natural substrates in cuttlefish, Sepia officinalis

The camouflaging abilities of cuttlefish (Sepia officinalis) are remarkable and well known. It is commonly believed that cuttlefish—although color blind—actively match various colors of their immediate surroundings, yet no quantitative data support this notion. We assembled several natural substrates chosen to evoke the three basic types of camouflaged body patterns that cuttlefish express (uniform/stipple, mottle, and disruptive) and measured the spectral reflectance of the camouflaged pattern and the respective background using a fiber optic spectrometer. We demonstrate that the reflectance spectra of cuttlefish skin patterns correlate closely with the spectra of these natural substrates. Since pigmented chromatophores play a key role in cephalopod color change, we also measured the spectral reflectance of individual cuttlefish chromatophores under the microscope, and confirm the results from a previous publication reporting three distinct colors of chromatophores (yellow, orange, and dark brown) on the animals’ dorsal side. Taken together, our results show that the color variations in substrate and animal skin can be very similar and that this may facilitate color match on natural substrates in the absence of color vision.