Decoupled evolution of ventral and dorsal scales in agamid lizards: ventral keels are associated with arboreality.

Arboreality has evolved in all major vertebrate lineages and is often associated with morphological adaptations and increased diversification concomitant with accessing novel niche space. In squamate reptiles, foot, claw, and tail morphology are well-studied adaptations shown to be associated with transitions to arboreality. Here, we examined a less well understood trait-the keeled scale-in relation to microhabitat, climate, and diversification dynamics across a diverse lizard radiation, Agamidae. We found that the ancestral agamid had keeled dorsal but not ventral scales; further, dorsal and ventral keels are evolutionarily decoupled. Ventral keeled scales evolved repeatedly in association with arboreality and may be advantageous in reducing wear or by promoting interlocking when climbing. We did not find an association between keeled scales and diversification, suggesting keels do not allow finer-scale microhabitat partitioning observed in other arboreal-associated traits. We additionally found a relationship between keeled ventral scales and precipitation in terrestrial species where we posit that the keels may function to reduce scale degradation. Our results suggest that keeled ventral scales facilitated transitions to arboreality across agamid lizards, and highlight a need for future studies that explore their biomechanical function in relation to microhabitat and climate.

Scale-based landmark for internal organs assessments of common carp (Cyprinus carpio) during digital radiography and ultrasonography imaging.

Medical imaging techniques such as digital radiography and ultrasonography are non-invasive and provide precise results for examining internal organs and structures within fish. Their effectiveness can be further enhanced by using body parts like scales as markers for the organs beneath them. This study utilized the number of scales as landmarks in digital radiography and ultrasonography to non-invasively evaluate the muscles, bones, and images of internal and reproductive organs of common carp (Cyprinus carpio). Digital radiography was performed in the dorsoventral and lateral views of the fish, whereas ultrasonography was conducted in longitudinal and transverse views on sequence scale numbers with brightness and colour Doppler-modes. Digital radiography of the common carp revealed the whole-body morphology, including the bony parts from the head, pectoral fins, dorsal fins, pelvic fins, anal fins, and vertebrae to the tail that appeared radiopaque. Internal organs were also observed, with the swim bladder and heart appeared radiolucent, while the intestines, liver, testes, and ovaries appeared radiopaque. Ultrasonography in brightness mode displayed the digestive organs, reproductive organs, and muscle thickness. Additionally, colour Doppler mode demonstrated blood flow within the heart's ventricle.

Digestion and absorption characteristics of iron-chelating silver carp scale collagen peptide and insights into their chelation mechanism.

Iron deficiency is widespread throughout the world, supplementing sufficient iron or improving the bioavailability of iron is the fundamental strategy to solve the problem of iron scarcity. Herein, we explored a new form of iron supplement, iron chelates of silver carp scales (SCSCP-Fe) were prepared from collagen peptide of silver carp scales (SCSCP) and FeCl2·4H2O, the effects of external environment and simulated gastrointestinal digestive environment on the stability of SCSCP-Fe and the structural changes of peptide iron chelates during digestion were investigated. The results of in vitro iron absorption promotion showed that the iron bioavailability of SCSCP-Fe was higher than that of FeSO4. Two potential high iron chelating peptides DTSGGYDEY (DY) and LQGSNEIEIR (LR) were screened and synthesized from the SCSCP sequence by molecular dynamics and LC-MS/MS techniques. The FTIR results displayed that the binding sites of DY and LR for Fe2+ were the carboxyl group, the amino group, and the nitrogen atom on the amide group on the peptide. ITC results indicated that the chelation reactions of DY and LR with Fe2+ were mainly dominated by electrostatic interactions, forming chelates in stoichiometric ratios of 1:2 and 1:1, respectively. Both DY and LR had a certain ability to promote iron absorption. The transport of DY-Fe chelate may be a combination of the three pathways: PepT1 vector pathway, cell bypass, and endocytosis, while LR-Fe chelate was dominated by bivalent metal ion transporters. This study is expected to provide theoretical reference and technical support for the high-value utilization of silver carp scales and the development of novel iron supplements.

Eco-friendly engineering of micro composite-based hydroxyapatite bio crystal and polyaniline for high removal of OG dye from wastewater: Adsorption mechanism and RSM@BBD optimization.

The presence of harmful substances such as dyes in water systems poses a direct threat to the quality of people's lives and other organisms living in the ecosystem. Orange G (OG) is considered a hazardous dye. The existing paper attempts to evaluate a low-cost adsorbent for the effective removal of OG dye. The developed adsorbent Polyaniline@Hydroxyapatite extracted from Cilus Gilberti fish Scale (PANI@FHAP) was elaborated through the application of the in situ chemical polymerization method to incorporate PANI on the surface of naturally extracted hydroxyapatite FHAP. The good synthesis of PANI@FHAP was evaluated through multiple techniques including X-ray diffraction (XRD), Scanning electron microscopy coupled with energy dispersive X-ray spectrometry (SEM/EDS), Fourier Transforms Infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA) coupled with thermal differential analysis (DTA) analysis. The results reveal a highly ordered disposition of PANI chains on FHAP, resulting in a well-coated FHAP in the PANI matrix. Furthermore, the presence of functional groups on the surface of PANI such as amine (-NH2) and imine (=NH) groups would facilitate the removal of OG dye from contaminated water. The adsorption of OG onto PANI@FHAP was conducted in batch mode and optimized through response surface methodology coupled with box-Behnken design (RSM/BBD) to investigate the effect of time, adsorbent dose, and initial concentration. The outcomes proved that OG adsorption follows a quadratic model (R2 = 0.989). The kinetic study revealed that the adsorption of OG fits the pseudo-second-order model. On the other hand, the isotherm study declared that the Freundlich model is best suited to the description of OG adsorption. For thermodynamic study, the adsorption of OG is spontaneous in nature and exothermic. Furthermore, the regeneration-reusability study indicates that PANI@FHAP could be regenerated and reused up to five successive cycles. Based on the FTIR spectrum of PANI@FHAP after OG adsorption, the mechanism governing OG adsorption is predominantly driven by π-π interaction, electrostatic interaction, and hydrogen bonding interactions. The obtained results suppose that PANI@FHAP adsorbent can be a competitive material in large-scale applications.

Spatio-temporal expression patterns of glycine-rich beta proteins and cysteine-rich beta proteins in setae development of Gekko japonicus.

BACKGROUND: Setae on the pad lamellae of the Japanese gecko Gekko japonicus (Schlegel, 1836), a vital epidermal derivative, are primarily composed of cornified beta-proteins (CBPs) and play a pivotal role in adhesion and climbing. The amino acid composition of CBPs might be a determining factor influencing their functional properties. However, the molecular mechanisms governed by CBP genes with diverse amino acid compositions in setae development remain unexplored. RESULTS: Based on RNA-seq analyses, this study confirmed that all G. japonicus CBPs (GjCBPs) are involved in setae formation. Cysteine-rich CBPs encoding genes (ge-cprp-17 to ge-cprp-26) and glycine-rich CBPs encoding genes (ge-gprp-17 to ge-gprp-22) were haphazardly selected, with quantitative real-time PCR revealing their expression patterns in embryonic pad lamellae and dorsal epidermis. It is inferred that glycine-rich CBPs are integral to the formation of both dorsal scales and lamellar setae, cysteine-rich CBPs are primarily associated with setae development. Additionally, fluorescence in situ hybridization revealed spatiotemporal differences in the expression of a glycine-rich CBP encoding gene (ge-gprp-19) and a cysteine-rich CBP encoding gene (ge-cprp-17) during dorsal scales and/or lamellar development. CONCLUSIONS: All 66 CBPs are involved in the formation of setae. Glycine-rich CBPs hold a significant role in the development of dorsal scales and lamellar setae, whereas most cysteine-rich CBPs appear to be essential components of G. japonicus setae. Even GjCBPs with similar amino acid compositions may play diverse functions. The clear spatio-temporal expression differences between the glycine-rich and cysteine-rich CBP encoding genes during epidermal scale and/or setae formation were observed. Embryonic developmental stages 39 to 42 emerged as crucial phases for setae development. These findings lay the groundwork for deeper investigation into the function of GjCBPs in the development of G. japonicus setae.

Effect and mechanism of natural composite hydrogel from fish scale intercellular matrix on diabetic chronic wound repair.

Diabetes mellitus is a chronic metabolic disease with prolonged low-grade inflammation and impaired cellular function, leading to poor wound healing. The treatment of diabetic wounds remains challenging due to the complex wound microenvironment. In view of the prominence of fish scales in traditional Chinese medicine and their wide application in modern medicine, we isolated the intercellular components in the scales of sea bass, obtained a natural composite hydrogel, fish scales gel (FSG), and applied it to diabetic chronic wounds. FSG was rich in collagen-like proteins, and possessed low-temperature gelation properties. In vitro, FSG was biocompatible and promoted fibroblast proliferation by approximately 40 %, endothelial cell migration by approximately 20 % and activated the M1 macrophages. In addition, FSG restored the function of fibroblasts and vascular endothelial cells damaged by high glucose. Importantly, FSG normalized the acute inflammatory response to impaired macrophages in a high-glucose microenvironment. Transcriptome analysis implies that this mechanism may involve enhanced cell signaling and cellular communication, improved sensitivity to cytokines, and activation of the TNF signaling pathway. Animal experiments confirmed that FSG significantly improved wound closure by approximately 15 % in diabetic rats, showing similar effects to acute wounds. In conclusion, the regulation of multiple cellular functions by FSG, especially the counterintuitive ability to induce acute inflammation, promoted diabetic wound healing and provides a novel therapeutic strategy for wound repair in diabetic patients.

The age and growth information of a ctenoid scale fossil from the Upper Cretaceous Nenjiang Formation in Songliao Basin, China.

The study of morphological characteristics and growth information in fish scales is a crucial component of modern fishery biological research, while it has been less studied in fossil materials. This paper presents a detailed morphological description and growth analysis of a fossil ctenoid scale obtained from the Upper Cretaceous Campanian lacustrine deposits in northeastern China. The morphological features of this fossil scale are well-preserved and consistent with the structures found in ctenoid scales of extant fish species and display prominent ring ornamentation radiating outward from the central focus, with grooves intersecting the rings. A comparative analysis of the morphological characteristics between the fossil ctenoid scale and those well-studied extant fish Mugilidae allows us to explore the applicability of modern fishery biological research methods to the field of fossil scales. The scale length, scale width, the vertical distance from the focus to the apex of the scale, and the total number of radii have been measured. The age of the fish that possessed this ctenoid scale has been estimated by carefully counting the annuli, suggesting an age equal to or more than seven years. The distribution of growth rings on the scale potentially reflects the warm paleoclimatic condition and fish-friendly paleoenvironment prevalent during that period. This paper, moreover, serves as a notable application of fishery biological methods in the examination of fossil materials.

Cellular structure of dinosaur scales reveals retention of reptile-type skin during the evolutionary transition to feathers.

Fossil feathers have transformed our understanding of integumentary evolution in vertebrates. The evolution of feathers is associated with novel skin ultrastructures, but the fossil record of these changes is poor and thus the critical transition from scaled to feathered skin is poorly understood. Here we shed light on this issue using preserved skin in the non-avian feathered dinosaur Psittacosaurus. Skin in the non-feathered, scaled torso is three-dimensionally replicated in silica and preserves epidermal layers, corneocytes and melanosomes. The morphology of the preserved stratum corneum is consistent with an original composition rich in corneous beta proteins, rather than (alpha-) keratins as in the feathered skin of birds. The stratum corneum is relatively thin in the ventral torso compared to extant quadrupedal reptiles, reflecting a reduced demand for mechanical protection in an elevated bipedal stance. The distribution of the melanosomes in the fossil skin is consistent with melanin-based colouration in extant crocodilians. Collectively, the fossil evidence supports partitioning of skin development in Psittacosaurus: a reptile-type condition in non-feathered regions and an avian-like condition in feathered regions. Retention of reptile-type skin in non-feathered regions would have ensured essential skin functions during the early, experimental stages of feather evolution.

Zebrafish Scale Regeneration In Toto and Ex Vivo Culture of Scales.

Skeletal diseases are often complex in their etiology and affect millions of people worldwide. Due to the aging population, there is a need for new therapeutics that could ease the burden on healthcare systems. As these diseases are complex, it is difficult and expensive to accurately model bone pathophysiology in a lab setting. The challenge for the field is to establish a cost-effective, biologically relevant platform for modeling bone disease that can be used to test potential therapeutic compounds. Such a platform should ideally allow dynamic visualization of cell behaviors of bone-building osteoblasts and bone-degrading osteoclasts acting in their mineralized matrix environment. Zebrafish are increasingly used as models due to the availability of genetic tools, including transgenic reporter lines, and the fact that some skeletal tissues (including the scales) remain translucent to adulthood, allowing dynamic imaging options. Since zebrafish scales have both osteoblasts and osteoclasts and are highly abundant, they provide an easily accessible and abundantly available resource of independent bone units. Moreover, once removed, adult zebrafish scales fully regenerate, therefore offering a way to study the spatiotemporal growth of mineralized tissue in vivo. Here, we detail protocols for harvesting and tracking the regeneration of the scales. Lastly, a protocol for stable culture of scales ex vivo for a week and following the healing response after controlled damage to the mineralized matrix of the scale over time is also presented.

The actin cytoskeleton plays multiple roles in structural colour formation in butterfly wing scales.

Vivid structural colours in butterflies are caused by photonic nanostructures scattering light. Structural colours evolved for numerous biological signalling functions and have important technological applications. Optically, such structures are well understood, however insight into their development in vivo remains scarce. We show that actin is intimately involved in structural colour formation in butterfly wing scales. Using comparisons between iridescent (structurally coloured) and non-iridescent scales in adult and developing H. sara, we show that iridescent scales have more densely packed actin bundles leading to an increased density of reflective ridges. Super-resolution microscopy across three distantly related butterfly species reveals that actin is repeatedly re-arranged during scale development and crucially when the optical nanostructures are forming. Furthermore, actin perturbation experiments at these later developmental stages resulted in near total loss of structural colour in H. sara. Overall, this shows that actin plays a vital and direct templating role during structural colour formation in butterfly scales, providing ridge patterning mechanisms that are likely universal across lepidoptera.

A diadectid skin impression and its implications for the evolutionary origin of epidermal scales.

Corneous skin appendages are not only common and diverse in crown-group amniotes but also present in some modern amphibians. This raises the still unresolved question of whether the ability to form corneous skin appendages is an apomorphy of a common ancestor of amphibians and amniotes or evolved independently in both groups. So far, there is no palaeontological contribution to the issue owing to the lack of keratin soft tissue preservation in Palaeozoic anamniotes. New data are provided by a recently discovered ichnofossil specimen from the early Permian of Poland that shows monospecific tetrapod footprints associated with a partial scaly body impression. The traces can be unambiguously attributed to diadectids and are interpreted as the globally first evidence of horned scales in tetrapods close to the origin of amniotes. Taking hitherto little-noticed scaly skin impressions of lepospondyl stem amniotes from the early Permian of Germany into account, the possibility has to be considered that the evolutionary origin of epidermal scales deeply roots among anamniotes.

Taxonomical use of scale ornamentation: Challenges by intraspecific and intra-individual variations in four adult specimens of Polypterus bichir.

Many actinopterygian fish groups, including fossil and extant polypteriforms and lepisosteiforms, fossil halecomorphs, and some basal teleosts, have stout bony scales covered by layers of ganoin-an enamel layer ornamented with minute tubercles. Ganoid scales preserve well as disarticulated remains and notably constitute most of the fossil record for polypteriform in both South America and Africa. Based on two variables (tubercle size and distance between tubercles), some authors reported that the ganoin tubercle ornamentation in these scales is constant within a species and differs between species and allows distinguishing species or at least groups of species. However, despite its promising potential for assessing polypteriform paleodiversity, this tool has remained unused, probably because the variables are not well defined, and intraspecific variation does not seem to have been considered. To address this gap, we aimed to test the intraspecific and intra-individual variation in the ornamentation of ganoid scales in the type species Polypterus bichir. We propose three different parameters to describe the tubercle ornamentation: the distance between contiguous tubercles centers, their density, and their relative spatial organization. With these parameters, we investigate the variation in ganoin ornamentation among four specimens and across different regions of the body. Our results show that the distribution of the tubercles is highly variable within a same species, regardless of the body region, and sometimes even between different sectors of a same scale. Moreover, the variation observed in P. bichir overlaps with the distribution described in the literature for several extant and fossil species. Thus far, the ornamentation of ganoid scales is not a reliable diagnostical feature for polypterids.

Mechanical, optical, chemical, and biological evaluations of fish scale-derived scaffold for corneal replacements: A systematic review.

Corneal blindness is commonly treated through corneal replacement with allogeneic corneal donors, which may face shortage. Regarding this issue, xenogeneic alternatives are explored. Fish scale-derived scaffolds (FSSs) are among the alternatives due to the lower risk of infection and abundant sources of raw materials. Unfortunately, the information about mechanical, optical, chemical, and biological performances of FSSs for corneal replacements is still scattered, as well as about the fabrication techniques. This study aims to gather scattered pieces of information about the mentioned performances and fabrication techniques of FSSs for corneal replacements. Sorted from four scientific databases and using the PRISMA checklist, eleven relevant articles are collected. FSSs are commonly fabricated using decellularization and decalcification processes, generating FSSs with parallel multilayers or crossed fibers with topographic microchannels. In the collected studies, similar mechanical properties of FSSs to native tissues are discovered, as well as good transparency, light remittance, but poorer refractive indexes than native tissues. Biological evaluations mostly discuss histology, cell proliferations, and immune responses on FSSs, while only a few studies examine the vascularization. No studies completed comprehensive evaluations on the four properties. The current progress of FSS developments demonstrates the potential of FSS use for corneal replacements.

Vertebrate mineralized tissues: A modular structural analysis.

Vertebrate mineralized tissues, present in bones, teeth and scales, have complex 3D hierarchical structures. As more of these tissues are characterized in 3D using mainly FIB SEM at a resolution that reveals the mineralized collagen fibrils and their organization into collagen fibril bundles, highly complex and diverse structures are being revealed. In this perspective we propose an approach to analyzing these tissues based on the presence of modular structures: material textures, pore shapes and sizes, as well as extents of mineralization. This modular approach is complimentary to the widely used hierarchical approach for describing these mineralized tissues. We present a series of case studies that show how some of the same structural modules can be found in different mineralized tissues, including in bone, dentin and scales. The organizations in 3D of the various structural modules in different tissues may differ. This approach facilitates the framing of basic questions such as: are the spatial relations between modular structures the same or similar in different mineralized tissues? Do tissues with similar sets of modules carry out similar functions or can similar functions be carried out using a different set of modular structures? Do mineralized tissues with similar sets of modules have a common developmental or evolutionary pathway? STATEMENT OF SIGNIFICANCE: 3D organization studies of diverse vertebrate mineralized tissues are revealing detailed, but often confusing details about the material textures, the arrangements of pores and differences in the extent of mineralization within a tissue. The widely used hierarchical scheme for describing such organizations does not adequately provide a basis for comparing these tissues, or addressing issues such as structural components thought to be characteristic of bone, being present in dermal tissues and so on. The classification scheme we present is based on identifying structural components within a tissue that can then be systematically compared to other vertebrate mineralized tissues. We anticipate that this classification approach will provide insights into structure-function relations, as well as the evolution of these tissues.

The anti-cancer effect and mechanism of animal scale-derived extract on malignant melanoma cells.

Melanoma is a type of cancer with abnormal proliferation of melanocytes and is one of the most diagnosed cancer types. In traditional Chinese medicine, pangolin scales have been used to treat various diseases, including human cancers. However, its efficacy has not been scientifically proven. Here we studied the anticancer effect and mechanism of pangolin scale extract (PSE) on melanoma cell lines using scientific approaches. Our cell viability assay shows that PSE exhibits up to approximately 50-80% inhibition on SK-MEL-103 and A375 melanoma cell lines. Mechanically, PSE inhibits melanoma cell proliferation, migration, and causes changes in cell morphology. The apoptosis assay showed a significant chromosomal condensation inside the PSE-treated melanoma cells. The sequencing and analysis of A375 melanoma cell transcriptomes revealed 3077 differentially expressed genes in the 6 h treatment group and 8027 differentially expressed genes in the 72 h treatment group. Transcriptome analysis suggests that PSE may cause cell cycle arrest in melanoma cells and promote apoptosis mainly by up-regulating the p53 signaling pathway and down-regulating the PI3K-Akt signaling pathway. In this study, the anticancer effect of PSE was demonstrated by molecular biological means. PSE shows a significant inhibition effect on melanoma cell proliferation and cell migration in vitro, causes cell cycle arrest and promotes apoptosis through p53 and PI3K-AKT pathways. This study provides better insights into the anti-cancer efficacy and underlying mechanism of PSE and a theoretical basis for mining anticancer compounds or the development of new treatments for melanoma in the future. It is worth noting that this study does not advocate the use of the pangolin scale for disease treatment, but only to confirm its usefulness from a scientific research perspective and to encourage subsequent research around the development of active compounds to replace pangolin scales to achieve the conservation of this endangered species.

A new elpistostegalian from the Late Devonian of the Canadian Arctic.

A fundamental gap in the study of the origin of limbed vertebrates lies in understanding the morphological and functional diversity of their closest relatives. Whereas analyses of the elpistostegalians Panderichthys rhombolepis, Tiktaalik roseae and Elpistostege watsoni have revealed a sequence of changes in locomotor, feeding and respiratory structures during the transition1-9, an isolated bone, a putative humerus, has controversially hinted at a wider range in form and function than now recognized10-14. Here we report the discovery of a new elpistostegalian from the Late Devonian period of the Canadian Arctic that shows surprising disparity in the group. The specimen includes partial upper and lower jaws, pharyngeal elements, a pectoral fin and scalation. This new genus is phylogenetically proximate to T. roseae and E. watsoni but evinces notable differences from both taxa and, indeed, other described tetrapodomorphs. Lacking processes, joint orientations and muscle scars indicative of appendage-based support on a hard substrate13, its pectoral fin shows specializations for swimming that are unlike those known from other sarcopterygians. This unexpected morphological and functional diversity represents a previously hidden ecological expansion, a secondary return to open water, near the origin of limbed vertebrates.

Mechanistic bottom-up estimation of passive drug absorption from the gastrointestinal tract: Comparison among primary cultured human intestinal cells, Caco-2 cells, artificial membrane, and animal scale-up.

OBJECTIVE: The fraction of drug absorbed (Fa) from the intestine is an important parameter to characterize the pharmacokinetics of a drug. We aimed to search for an experimental system that provides the best parameters for estimating the effective permeability (Peff) used for the bottom-up prediction of Fa. MATERIALS AND METHODS: The absorption kinetics of 12 passively absorbed drugs were simulated by a compartment absorption transit (CAT) model using absorption parameters from four different experimental systems: human intestinal epithelial cell (HIEC) monolayer, Caco-2 monolayer, parallel artificial membrane permeability assay (PAMPA), and in situ rat intestinal perfusion. All absorption parameters were obtained from the literature. The in vitro apparent permeability coefficient (Papp) and rat in situ Peff were converted to human Peff using a bottom-up approach for each region, based on the morphological features of the human intestine. The simulated Fa values were compared to the respective observed values. Furthermore, plasma concentration profiles of the drugs were simulated by convolution using the time-course of the absorption rate simulated using the Peff values calculated from the HIEC Papp. RESULTS: The Fa values were best predicted by using the Peff values calculated from HEIC, within a 1.3-fold range of observed Fa in 11 out of 12 drugs. The simulated Cmax values of pharmacokinetic simulation using HIEC Papp fell within a 1.5-fold range of observed values for all the drugs examined. CONCLUSION: The HIEC monolayer was identified as the most suitable permeation parameter for estimating Fa and Cmax using a morphological feature-based bottom-up approach.

Single-cell transcriptomics defines keratinocyte differentiation in avian scutate scales.

The growth of skin appendages, such as hair, feathers and scales, depends on terminal differentiation of epidermal keratinocytes. Here, we investigated keratinocyte differentiation in avian scutate scales. Cells were isolated from the skin on the legs of 1-day old chicks and subjected to single-cell transcriptomics. We identified two distinct populations of differentiated keratinocytes. The first population was characterized by mRNAs encoding cysteine-rich keratins and corneous beta-proteins (CBPs), also known as beta-keratins, of the scale type, indicating that these cells form hard scales. The second population of differentiated keratinocytes contained mRNAs encoding cysteine-poor keratins and keratinocyte-type CBPs, suggesting that these cells form the soft interscale epidermis. We raised an antibody against keratin 9-like cysteine-rich 2 (KRT9LC2), which is encoded by an mRNA enriched in the first keratinocyte population. Immunostaining confirmed expression of KRT9LC2 in the suprabasal epidermal layers of scutate scales but not in interscale epidermis. Keratinocyte differentiation in chicken leg skin resembled that in human skin with regard to the transcriptional upregulation of epidermal differentiation complex genes and genes involved in lipid metabolism and transport. In conclusion, this study defines gene expression programs that build scutate scales and interscale epidermis of birds and reveals evolutionarily conserved keratinocyte differentiation genes.

Morphology and distribution of scales, dermal ossifications, and other non-feather integumentary structures in non-avialan theropod dinosaurs.

Modern birds are typified by the presence of feathers, complex evolutionary innovations that were already widespread in the group of theropod dinosaurs (Maniraptoriformes) that include crown Aves. Squamous or scaly reptilian-like skin is, however, considered the plesiomorphic condition for theropods and dinosaurs more broadly. Here, we review the morphology and distribution of non-feathered integumentary structures in non-avialan theropods, covering squamous skin and naked skin as well as dermal ossifications. The integumentary record of non-averostran theropods is limited to tracks, which ubiquitously show a covering of tiny reticulate scales on the plantar surface of the pes. This is consistent also with younger averostran body fossils, which confirm an arthral arrangement of the digital pads. Among averostrans, squamous skin is confirmed in Ceratosauria (Carnotaurus), Allosauroidea (Allosaurus, Concavenator, Lourinhanosaurus), Compsognathidae (Juravenator), and Tyrannosauroidea (Santanaraptor, Albertosaurus, Daspletosaurus, Gorgosaurus, Tarbosaurus, Tyrannosaurus), whereas dermal ossifications consisting of sagittate and mosaic osteoderms are restricted to Ceratosaurus. Naked, non-scale bearing skin is found in the contentious tetanuran Sciurumimus, ornithomimosaurians (Ornithomimus) and possibly tyrannosauroids (Santanaraptor), and also on the patagia of scansoriopterygids (Ambopteryx, Yi). Scales are surprisingly conservative among non-avialan theropods compared to some dinosaurian groups (e.g. hadrosaurids); however, the limited preservation of tegument on most specimens hinders further interrogation. Scale patterns vary among and/or within body regions in Carnotaurus, Concavenator and Juravenator, and include polarised, snake-like ventral scales on the tail of the latter two genera. Unusual but more uniformly distributed patterning also occurs in Tyrannosaurus, whereas feature scales are present only in Albertosaurus and Carnotaurus. Few theropods currently show compelling evidence for the co-occurrence of scales and feathers (e.g. Juravenator, Sinornithosaurus), although reticulate scales were probably retained on the mani and pedes of many theropods with a heavy plumage. Feathers and filamentous structures appear to have replaced widespread scaly integuments in maniraptorans. Theropod skin, and that of dinosaurs more broadly, remains a virtually untapped area of study and the appropriation of commonly used techniques in other palaeontological fields to the study of skin holds great promise for future insights into the biology, taphonomy and relationships of these extinct animals.

Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle.

We developed a multiphoton imaging method to capture neural structure and activity in behaving flies through the intact cuticle. Our measurements showed that the fly head cuticle has surprisingly high transmission at wavelengths >900nm, and the difficulty of through-cuticle imaging is due to the air sacs and/or fat tissue underneath the head cuticle. By compressing or removing the air sacs, we performed multiphoton imaging of the fly brain through the intact cuticle. Our anatomical and functional imaging results show that 2- and 3-photon imaging are comparable in superficial regions such as the mushroom body, but 3-photon imaging is superior in deeper regions such as the central complex and beyond. We further demonstrated 2-photon through-cuticle functional imaging of odor-evoked calcium responses from the mushroom body γ-lobes in behaving flies short term and long term. The through-cuticle imaging method developed here extends the time limits of in vivo imaging in flies and opens new ways to capture neural structure and activity from the fly brain.