Vertebrate Integument Research Articles

Preservation of corneous β-proteins in Mesozoic feathers.

Fossil proteins are valuable tools in evolutionary biology. Recent technological advances and better integration of experimental methods have confirmed the feasibility of biomolecular preservation in deep time, yielding new insights into the timing of key evolutionary transitions. Keratins (formerly α-keratins) and corneous β-proteins (CBPs, formerly β-keratins) are of particular interest as they define tissue structures that underpin fundamental physiological and ecological strategies and have the potential to inform on the molecular evolution of the vertebrate integument. Reports of CBPs in Mesozoic fossils, however, appear to conflict with experimental evidence for CBP degradation during fossilization. Further, the recent model for molecular modification of feather chemistry during the dinosaur-bird transition does not consider the relative preservation potential of different feather proteins. Here we use controlled taphonomic experiments coupled with infrared and sulfur X-ray spectroscopy to show that the dominant β-sheet structure of CBPs is progressively altered to α-helices with increasing temperature, suggesting that (α-)keratins and α-helices in fossil feathers are most likely artefacts of fossilization. Our analyses of fossil feathers shows that this process is independent of geological age, as even Cenozoic feathers can comprise primarily α-helices and disordered structures. Critically, our experiments show that feather CBPs can survive moderate thermal maturation. As predicted by our experiments, analyses of Mesozoic feathers confirm that evidence of feather CBPs can persist through deep time.

Sediment-encased pressure–temperature maturation experiments elucidate the impact of diagenesis on melanin-based fossil color and its paleobiological implications

AbstractMelanin pigments are central to colors and patterns in modern vertebrate integuments, which inform upon ecological and behavioral strategies like crypsis, aposematism, and sociosexual selection. Over the last decade, melanin has emerged as a valuable tool for predicting color in exceptionally preserved fossil feathers and subsequent testing of paleobiological hypotheses in long-extinct dinosaurs and birds. Yet much remains to be learned about melanin stability, diagenetic alterations to melanin chemistry, and their implications for “paleocolor reconstruction.” Pressure–temperature maturation experiments with modern feathers offer a way to examine these topics but have mostly been conducted in closed-system capsules or open-system aluminum foil. Both methods have operational limitations and do not consider the filtering effect of porous sediment matrices on thermally labile chemical groups versus stable ones during natural fossilization. We use sediment-encased maturation to resolve this issue and demonstrate replication of organic preservation of melanin highly comparable to compression fossils. Our experiments, coupled with time-of-flight secondary ion mass spectrometry, show predictable volatilization of N/S-bearing molecules and increased melanin cross-linking with elevated temperatures. We also suggest that eumelanin is more stable compared with pheomelanin at higher temperatures, explaining why eumelanic colors (black, dark brown, iridescent) are better preserved in fossils than pheomelanic ones (reddish brown). Furthermore, we propose that proteins preferentially undergo hydrolysis more so than forming N-heterocycles in selectively open systems analogous to natural matrices. Thus, we conclude that melanin pigments and not diagenetically altered protein remnants are the key players in promoting fossilization of soft tissues like feathers.

Morpho-structural adaptations of the integument in different aquatic organisms

The integument acts as a barrier to protect the body from harmful pathogenic infectious agents, parasites, UV rays, trauma, and germs. The integument of invertebrates and vertebrates are structurally different: while invertebrates usually have a simple monolayer epidermis frequently covered by mucus, cuticles, or mineralized structures, vertebrates possess a multilayered epidermis with several specialized cells. This study aims to describe by morphological, histological, and immunohistochemical analyses, the morpho-structural adaptations throughout evolution of the integument of gastropod Aplysia depilans (Gmelin, 1791), ascidian Styela plicata (Lesuer, 1823), myxine hagfish Eptatretus cirrhatus (Forster, 1801) and teleost Heteropneustes fossilis (Bloch, 1794) for the first time, with special reference to sensory epidermal cells. Different types of cells could be identified that varied according to the species; including mucous cells, serous glandular cells, clavate cells, club cells, thread cells, and support cells. In all integuments of the specimens analyzed, sensory solitary cells were identified in the epidermis, immunoreactive to serotonin and calbindin. Our study provided an essential comparison of integuments, adding new information about sensory epidermal cells phylogenetic conservation and on the structural changes that invertebrates and vertebrates have undergone during evolution.

Introduction to the Development of Skin in Vertebrates

The integument of vertebrates is a complex and large organ positioned at the interface with the aquatic or terrestrial environment, and is derived from the embryonic ectoderm (epidermis) and mesoderm (dermis and hypodermis) [...].

Molecular parallelisms between pigmentation in the avian iris and the integument of ectothermic vertebrates.

Birds exhibit striking variation in eye color that arises from interactions between specialized pigment cells named chromatophores. The types of chromatophores present in the avian iris are lacking from the integument of birds or mammals, but are remarkably similar to those found in the skin of ectothermic vertebrates. To investigate molecular mechanisms associated with eye coloration in birds, we took advantage of a Mendelian mutation found in domestic pigeons that alters the deposition of yellow pterin pigments in the iris. Using a combination of genome-wide association analysis and linkage information in pedigrees, we mapped variation in eye coloration in pigeons to a small genomic region of ~8.5kb. This interval contained a single gene, SLC2A11B, which has been previously implicated in skin pigmentation and chromatophore differentiation in fish. Loss of yellow pigmentation is likely caused by a point mutation that introduces a premature STOP codon and leads to lower expression of SLC2A11B through nonsense-mediated mRNA decay. There were no substantial changes in overall gene expression profiles between both iris types as well as in genes directly associated with pterin metabolism and/or chromatophore differentiation. Our findings demonstrate that SLC2A11B is required for the expression of pterin-based pigmentation in the avian iris. They further highlight common molecular mechanisms underlying the production of coloration in the iris of birds and skin of ectothermic vertebrates.

Cretaceous amniote integuments recorded through a taphonomic process unique to resins

Fossil records of vertebrate integuments are relatively common in both rocks, as compressions, and amber, as inclusions. The integument remains, mainly the Mesozoic ones, are of great interest due to the panoply of palaeobiological information they can provide. We describe two Spanish Cretaceous amber pieces that are of taphonomic importance, one bearing avian dinosaur feather remains and the other, mammalian hair. The preserved feather remains originated from an avian dinosaur resting in contact with a stalactite-shaped resin emission for the time it took for the fresh resin to harden. The second piece shows three hair strands recorded on a surface of desiccation, with the characteristic scale pattern exceptionally well preserved and the strands aligned together, which can be considered the record of a tuft. These assemblages were recorded through a rare biostratinomic process we call “pull off vestiture” that is different from the typical resin entrapment and embedding of organisms and biological remains, and unique to resins. The peculiarity of this process is supported by actualistic observations using sticky traps in Madagascar. Lastly, we reinterpret some exceptional records from the literature in the light of that process, thus bringing new insight to the taphonomic and palaeoecological understanding of the circumstances of their origins.

On the Potential Role of the Neural Crest Cells in Integrating Pigmentation Into Behavioral and Physiological Syndromes

Melanin pigments colour the integument of vertebrates with shades of rufous, grey, and black. In numerous wild species, melanin-based colouration associates to different behavioural, physiological, and morphological traits, yet the proximate basis of such associations remains largely unknown. The neural crest/domestication syndrome hypothesis (Wilkins et al. 2014) proposes that correlated changes in pigmentary, behavioural, physiological, and morphological traits of domestic species result from deficiencies affecting the number or development of neural crest cells in the embryo. Here, we review to what extent the neural crest cells might also explain the associations observed between melanin-based colouration and distinct traits in wild vertebrate populations. Genes involved in the development of the neural crest explain colour differences in some vertebrate species, particularly, in cichlid fishes. Cells that originate from the embryo’s neural crest cells play functions that have been previously associated with melanin-based colouration in wild vertebrates and we propose the potential mechanisms through which the association of melanin-based colouration with other traits can be the consequence of having a common origin at the neural crest cells. We encourage considering the influence of the neural crest as a new avenue of research, integrating developmental biology and evolutionary ecology to better understand why and how melanin-based colouration of vertebrates become associated to multiple phenotypic aspects.

The Vertebrate Integument: Origin and Evolution, Volume 1 The Vertebrate Integument: Structure, Design and Function, Volume 2

The Vertebrate Integument: Origin and Evolution, Volume 1 The Vertebrate Integument: Structure, Design and Function, Volume 2

New occurrences of fossilized feathers: systematics and taphonomy of the Santana Formation of the Araripe Basin (Cretaceous), NE, Brazil.

Here we describe three fossil feathers from the Early Cretaceous Santana Formation of the Araripe Basin, Brazil. Feathers are the most complex multiform vertebrate integuments; they perform different functions, occurring in both avian and non-avian dinosaurs. Despite their rarity, fossil feathers have been found across the world. Most of the Brazilian feather fossil record comes from the Santana Formation. This formation is composed of two members: Crato (lake) and Romualdo (lagoon); both of which are predominantly reduced deposits, precluding bottom dwelling organisms, resulting in exceptional preservation of the fossils. Despite arid and hot conditions during the Cretaceous, life teemed in the adjacency of this paleolake. Feathered non-avian dinosaurs have not yet been described from the Crato Member, even though there are suggestions of their presence in nearby basins. Our description of the three feathers from the Crato laminated limestone reveals that, despite the small sample size, they can be referred to coelurosaurian theropods. Moreover, based on comparisons with extant feather morphotypes they can be identified as one contour feather and two downy feathers. Despite their rareness and low taxonomic potential, fossilized feathers can offer insights about the paleobiology of its owners and the paleoecology of the Araripe Basin.

The vertebrate integument, v.1: origin and evolution

The vertebrate integument, v.1: origin and evolution

Adaptive evolution of Hoxc13 genes in the origin and diversification of the vertebrate integument.

The problem of origination and diversification of integument derivatives in vertebrates is still a challenge. The homeobox (Hox) genes Hoxc13 control integument formation in vertebrate. Hoxc13 show strong expression in the integument development, are highly conserved across vertebrates, and show mutations that are associated with skin and appendages. To test whether the evolution of the integument is associated with positive selection or relaxation of Hoxc13, we obtained these genes in a wide range of vertebrates. In Hoxc13, we found evidence of diversifying selection after speciation during the origin of vertebrates. In addition, we found the glycine-rich regions in Hoxc13 protein in mammals, but not among non-mammalian taxa. Our results strongly implicate that Hoxc13 genes could have played an important role in the evolution of integument structure.

Bird population declines due to radiation exposure at Chernobyl are stronger in species with pheomelanin-based coloration

Eumelanin and pheomelanin are the most common pigments providing color to the integument of vertebrates. While pheomelanogenesis requires high levels of a key intracellular antioxidant (glutathione, GSH), eumelanogenesis is inhibited by GSH. This implies that species that possess the molecular basis to produce large amounts of pheomelanin might be more limited in coping with environmental conditions that generate oxidative stress than species that produce eumelanin. Exposure to ionizing radiation produces free radicals and depletes antioxidant resources. GSH is particularly susceptible to radiation, so that species with large proportions of pheomelanic integument may be limited by the availability of GSH to combat oxidative stress and may thus suffer more from radiation effects. We tested this hypothesis in 97 species of birds censused in areas with varying levels of radioactive contamination around Chernobyl. After controlling for the effects of carotenoid-based color, body mass and similarity among taxa due to common phylogenetic descent, the proportion of pheomelanic plumage was strongly negatively related to the slope estimates of the relationship between abundance and radiation levels, while no effect of eumelanic color proportion was found. This represents the first report of an effect of the expression of melanin-based coloration on the capacity to resist the effects of ionizing radiation. Population declines were also stronger in species that exhibit carotenoid-based coloration and have large body mass. The magnitude of population declines had a relatively high phylogenetic signal, indicating that certain groups of birds, especially non-corvid passeriforms, are particularly susceptible to suffer from the effects of radioactive contamination due to phylogenetic inertia.

Calcified Integumentary Structures in Anurans

Although often overlooked, the integument of many vertebrates is reinforced by skeletal elements. In anurans these calcified structures include osteoderms, bone‐rich elements, and the lamina calcarea, a calcified, acellular layer that lacks collagen. We investigated the histology of anuran integument with a special focus on taxa developing skeletal elements. When present, the lamina calcarea forms a thin but discrete horizon at the interface between the superficial and deep dermis. This layer is rich in glycosaminoglycans (GAG) and is structurally unlike bone and cartilage. In contrast, anuran osteoderms are composed of cellular bone with limited GAG. A partial ontogenetic series of the tree frog Phyllomedusa bicolor provides the first details of anuran osteoderms development. In P. bicolor osteoderm development begins as a cluster of small calcified centres localized over the head. With continued body growth, osteoderms become larger in size and become distributed across the dorsal and ventral surfaces of the body, and across the limbs. The leaf litter frog Brachytarsophyrys cariensis is unusual in developing both osteoderms (on the dorsal body surface) and a lamina calcarea (on the ventral body surface). Our data shows that anuran integument has the ability to form skeletal elements and suggests that these elements arose in multiple anuran lineages.Grant Funding Source: NSERC

Dermatology in the Darwin anniversary. Part 1: Evolution of the integument

The present review highlights the development of the integument and its adnexa from the primitive metazoans to man. The different stages of development represent independent, partially convergent evolutions rather than a continuous evolutionary line. The epidermis of the invertebrates (sponges, cnidaria, worms, echinoderms and arthropods) always consists of one layer of pluripotent cells. The barrier function of the integument at this level is achieved with physico-chemical barriers, toxin production, fortification of the epidermis in the form of a cuticula, a syncytium or a neodermis. The lower vertebrates (cyclostoma, fishes and amphibians) have a stratified epidermis harboring many secretory cells. In terrestrial amphibians the outermost cell layer of the epidermis is cornified, and the secretory cells are relocated in the dermis. Terminal differentiation and cornification of the epidermis in the birds and mammals result in a more uniform shape of the epithelium. Stem cells are now restricted to some basal regions of the epithelium. In the mammals the glands are located in the deeper layers of the skin. In contrast to other vertebrate integuments the human skin does not possess specialized structures such as feathers, scales or coats. However, waiving specialization allows for unique universality of the human skin compared to other vertebrates.

First investigation of the collagen D -band ultrastructure in fossilized vertebrate integument

The ultrastructure of dermal fibres of a 200Myr thunniform ichthyosaur, Ichthyosaurus, specifically the 67nm axial repeat D-banding of the fibrils, which characterizes collagen, is presented for the first time by means of scanning electron microscopy (SEM) analysis. The fragment of material investigated is part of previously described fossilized skin comprising an architecture of layers of oppositely oriented fibre bundles. The wider implication, as indicated by the extraordinary quality of preservation, is the robustness of the collagen molecule at the ultrastructural level, which presumably contributed to its survival during the initial processes of decomposition prior to mineralization. Investigation of the elemental composition of the sample by SEM-energy dispersive X-ray spectroscopy indicates that calcite and phosphate played important roles in the rapid mineralization and fine replication of the collagen fibres and fibrils. The exceedingly small sample used in the investigation and high level of information achieved indicate the potential for minimal damage to prized museum specimens; for example, ultrastructural investigations by SEM may be used to help resolve highly contentious questions, for example, 'protofeathers' in the Chinese dinosaurs.

Abundant expression of SPARC, birc2 and boca mRNAs in goldfish scale

The existence of scales in teleost is related to the protective functions of the vertebrate integument and each scale is considered to be a counterpart of hair and/or nail. However, the scales can contain about 20% of the total body Ca so that they also appear to represent an internal Ca reservoir. Thus, in order to confirm whether scales are available for calcified tissue research or keratinized tissue research, characteristics of cells in the scale were examined by identifying differentially expressed genes. Subtraction cloning was subjected to scale ‐ gill (SG) and gill ‐ scale (GS). Six clones from SG and 4 from GS were isolated. Among them, SG1, SG2, and SG3 showed the high homology to SPARC, baculoviral IAP repeat containing 2 (birc2), and bcl2‐related ovarian killer a (boka), respectively. In situ hybridization confirmed birc2 and boca expression in flat cells along the ridge. Birc2 also expressed in large cells between the ridges. SPARC expressed in flat and small cells between the ridges. Although all the cells in the scale were designated as scleroblasts, there were several types of cells with different gene expression. Co‐expression of birc2 and boka along the ridge suggested that anti‐apoptotic mechanism worked to maintain the ridges. Since the cells did not always express SPARC, Ca homeostasis and/or osteocytic manifestation seemed to be maintained by the limited number of the cells.

Water relations of tetrapod integument

The vertebrate integument represents an evolutionary compromise between the needs for mechanical protection and those of sensing the environment and regulating the exchange of materials and energy. Fibrous keratins evolved as a means of strengthening the integument while simultaneously providing a structural support for lipids, which comprise the principal barrier to cutaneous water efflux in terrestrial taxa. Whereas lipids are of fundamental importance to water barriers, the efficacy of these barriers depends in many cases on structural features that enhance or maintain the integrity of function. Amphibians are exceptional among tetrapods in having very little keratin and a thin stratum corneum. Thus, effective lipid barriers that are present in some specialized anurans living in xeric habitats are external to the epidermis, whereas lipid barriers of amniotes exist as a lipid-keratin complex within the stratum corneum. Amphibians prevent desiccation of the epidermis and underlying tissues either by evaporating water from a superficial aqueous film, which must be replenished, or by shielding the stratum corneum with superficial lipids. Water barrier function in vertebrates generally appears to be relatively fixed, although various species have ;plasticity' to adjust the barrier effectiveness facultatively. While it is clear that both phenotypic plasticity and genetic adaptation can account for covariation between environment and skin resistance to water efflux, studies of the relative importance of these two phenomena are few. Fundamental mechanisms for adjusting the skin water barrier include changes in barrier thickness, composition and physicochemical properties of cutaneous lipids, and/or geometry of the barrier within the epidermis. While cutaneous lipids have been studied extensively in the contexts of disease and cosmetics, relatively little is known about the processes of permeability barrier ontogenesis related to adaptation and environment. Advances in such knowledge have didactic significance for understanding vertebrate evolution as well as practical application to clinical dermatology.

Dermal collagen organization in Bufo ictericus and in Rana catesbeiana integument (Anuran, Amphibian) under the evaluation of laser confocal microscopy

Collagen structural organization plays an important role in the mechanical property of the vertebrate integument. Bufo ictericus and Rana catesbeiana integument was investigated by light microscopy and laser confocal microscopy. Collagenous elements of the dermis were statistical analyzed. The integument is formed by the keratinized squamous stratified epidermis supported by the dermis that is subdivided into the spongious layer with a loose arrangement, and the compact layer formed by collagenous fibers arranged compactly in a criss-crossed manner. Thick collagenous columns have a perpendicular trajectory, and are formed by the assembling of alternating collagenous lamellae in both animals. Short intercolumns of collagenous fibrils connecting collagenous lamellae obliquely or transversally are observed in R. catesbeiana dorsal integument. The present study provides evidences that B. ictericus and R. catesbeiana integument has well-organized compact dermis, constituted by collagenous lamellae in a plywood manner. The integument organization is in contrast to the literature in some aspects. This dermal arrangement is important to the biomechanical property of both anuran integuments.

Plasticity of the water barrier in vertebrate integument

Water barrier function in vertebrates is typically relatively fixed and characteristic of species. However, both plasticity and genetic adaptation can account for covariation between environment and resistance to transepidermal water passage ( R s). The capacity to adjust R s may involve phenotypic plasticity, acclimation, or developmental plasticity, and fundamental mechanisms include changes in barrier thickness, composition and physicochemical properties of cutaneous lipids, and/or geometry of the barrier within the epidermis. In amniotes, lipid barriers are structured within keratin complexes that, while important to efficacy of function, appear to constrain the possibilities and time course of plastic responses. Studies of the relative importance of plastic responses and genetic variation are few and inconclusive.

The dinosaurian origin of feathers: perspectives from dolphin (Cetacea) collagen fibers.

The early origin of birds is a hotly disputed debate and may be broadly framed as a conflict between paleontologists and ornithologists. The paleontological emphasis has shifted from Archaeopteryx and its origins to recent finds of Cretaceous birds and "feathered" dinosaurs from China. The identification of alleged feathers has, however, relied principally on the visual image. Some workers have interpreted these integumentary structures as collagen fibers. To test the latter hypothesis, using light microscopy, collagen from the hypodermis (blubber) and subdermal connective tissue sheath was examined from a dolphin that had been buried for a year as part of an experiment. Within the blubber, toward the central thicker parts of the material, the collagen fibers had compacted and the three-dimensional latticework of normal blubber had more or less collapsed. Chromatographic analysis of the blubber revealed pronounced oxidation of the unsaturated lipids, probably accounting for the collapse of the latticework. Fibers normally bound together in bundles became separated into individual fibers or smaller bundles by degradation of the glue-like substance binding them together. These degraded collagen fibers show, in many instances, feather-like patterns, strikingly reminiscent of many of those identified as either "protofeathers" or "modern" feathers in dromaeosaurid dinosaurs. The findings throw serious doubt on the virtually complete reliance on visual image by supporters of the feathered dinosaur thesis and emphasize the need for more rigorous methods of identification using modern feathers as a frame of reference. Since collagen is the main fiber type found in most supporting tissues, the results have wide implications regarding the degradation and fossilization of vertebrate integument, such as that of the ichthyosaurs, dinosaurs and birds.