Structure and functions of keratin proteins in simple, stratified, keratinized and cornified epithelia.

Historically, the term 'keratin' stood for all of the proteins extracted from skin modifications, such as horns, claws and hooves. Subsequently, it was realized that this keratin is actually a mixture of keratins, keratin filament-associated proteins and other proteins, such as enzymes. Keratins were then defined as certain filament-forming proteins with specific physicochemical properties and extracted from the cornified layer of the epidermis, whereas those filament-forming proteins that were extracted from the living layers of the epidermis were grouped as 'prekeratins' or 'cytokeratins'. Currently, the term 'keratin' covers all intermediate filament-forming proteins with specific physicochemical properties and produced in any vertebrate epithelia. Similarly, the nomenclature of epithelia as cornified, keratinized or non-keratinized is based historically on the notion that only the epidermis of skin modifications such as horns, claws and hooves is cornified, that the non-modified epidermis is a keratinized stratified epithelium, and that all other stratified and non-stratified epithelia are non-keratinized epithelia. At this point in time, the concepts of keratins and of keratinized or cornified epithelia need clarification and revision concerning the structure and function of keratin and keratin filaments in various epithelia of different species, as well as of keratin genes and their modifications, in view of recent research, such as the sequencing of keratin proteins and their genes, cell culture, transfection of epithelial cells, immunohistochemistry and immunoblotting. Recently, new functions of keratins and keratin filaments in cell signaling and intracellular vesicle transport have been discovered. It is currently understood that all stratified epithelia are keratinized and that some of these keratinized stratified epithelia cornify by forming a Stratum corneum. The processes of keratinization and cornification in skin modifications are different especially with respect to the keratins that are produced. Future research in keratins will provide a better understanding of the processes of keratinization and cornification of stratified epithelia, including those of skin modifications, of the adaptability of epithelia in general, of skin diseases, and of the changes in structure and function of epithelia in the course of evolution. This review focuses on keratins and keratin filaments in mammalian tissue but keratins in the tissues of some other vertebrates are also considered.

The structure of the cornified claw sheath in the domesticated cat (Felis catus): implications for the claw-shedding mechanism and the evolution of cornified digital end organs.

The morphology of cornified structures is notoriously difficult to analyse because of the extreme range of hardness of their component tissues. Hence, a correlative approach using light microscopy, scanning electron microscopy, three-dimensional reconstructions based on x-ray computed tomography data, and graphic modeling was applied to study the morphology of the cornified claw sheath of the domesticated cat as a model for cornified digital end organs. The highly complex architecture of the cornified claw sheath is generated by the living epidermis that is supported by the dermis and distal phalanx. The latter is characterized by an ossified unguicular hood, which overhangs the bony articular base and unguicular process of the distal phalanx and creates an unguicular recess. The dermis covers the complex surface of the bony distal phalanx but also creates special structures, such as a dorsal dermal papilla that points distally and a curved ledge on the medial and lateral sides of the unguicular process. The hard-cornified external coronary horn and proximal cone horn form the root of the cornified claw sheath within the unguicular recess, which is deeper on the dorsal side than on the medial and lateral sides. As a consequence, their rate of horn production is greater dorsally, which contributes to the overall palmo-apical curvature of the cornified claw sheath. The external coronary and proximal cone horn is worn down through normal use as it is pushed apically. The hard-cornified apical cone horn is generated by the living epidermis enveloping the base and free part of the dorsal dermal papilla. It forms nested horn cones that eventually form the core of the hardened tip of the cornified claw. The sides of the cornified claw sheath are formed by the newly described hard-cornified blade horn, which originates from the living epidermis located on the slanted face of the curved ledge. As the blade horn is moved apically, it entrains and integrates the hard-cornified parietal horn on its internal side. It is covered by the external coronary and proximal cone horn on its external side. The soft-cornified terminal horn extends distally from the parietal horn and covers the dermal claw bed at the tip of the uniguicular process, thereby filling the space created by the converging apical cone and blade horn. The soft-cornified sole horn fills the space between the cutting edges of blade horn on the palmar side of the cornified claw sheath. The superficial soft-cornified perioplic horn is produced on the internal side of the unguicular pleat, which surrounds the root of the cornified claw sheath. The shedding of apical horn caps is made possible by the appearance of microcracks in the superficial layers of the external coronary and proximal cone horn in the course of deformations of the cornified claw sheath, which is subjected to tensile forces during climbing or prey catching. These microcracks propagate tangentially through the coronary horn and do not injure the underlying living epidermal and dermal tissues. This built-in shedding mechanism maintains sharp claw tips and ensures the freeing of the claws from the substrate.

3D MRI Modeling of Thin and Spatially Complex Soft Tissue Structures without Shrinkage: Lamprey Myosepta as an Example.

3D imaging techniques enable the nondestructive analysis and modeling of complex structures. Among these, MRI exhibits good soft tissue contrast, but is currently less commonly used for nonclinical research than X-ray CT, even though the latter requires contrast-staining that shrinks and distorts soft tissues. When the objective is the creation of a realistic and complete 3D model of soft tissue structures, MRI data are more demanding to acquire and visualize and require extensive post-processing because they comprise noncubic voxels with dimensions that represent a trade-off between tissue contrast and image resolution. Therefore, thin soft tissue structures with complex spatial configurations are not always visible in a single MRI dataset, so that standard segmentation techniques are not sufficient for their complete visualization. By using the example of the thin and spatially complex connective tissue myosepta in lampreys, we developed a workflow protocol for the selection of the appropriate parameters for the acquisition of MRI data and for the visualization and 3D modeling of soft tissue structures. This protocol includes a novel recursive segmentation technique for supplementing missing data in one dataset with data from another dataset to produce realistic and complete 3D models. Such 3D models are needed for the modeling of dynamic processes, such as the biomechanics of fish locomotion. However, our methodology is applicable to the visualization of any thin soft tissue structures with complex spatial configurations, such as fasciae, aponeuroses, and small blood vessels and nerves, for clinical research and the further exploration of tensegrity. Anat Rec, 301:1745-1763, 2018. © 2018 Wiley Periodicals, Inc.

The Human Shoulder Suspension Apparatus: A Causal Explanation for Bilateral Asymmetry and a Fresh Look at the Evolution of Human Bipedality.

The combination of large mastoid processes and clavicles is unique to humans, but the biomechanical and evolutionary significance of their special configuration is poorly understood. As part of the newly conceptualized shoulder suspension apparatus, the mastoid processes and clavicles are shaped by forces exerted by the musculo-fascial components of the cleidomastoid and clavotrapezius muscles as they suspend the shoulders from the head. Because both skeletal elements develop during infancy in tandem with the attainment of an upright posture, increased manual dexterity, and the capacity for walking, we hypothesized that the same forces would have shaped them as the shoulder suspension apparatus evolved in ancestral humans in tandem with an upright posture, increased manual dexterity, and bipedality with swinging arms. Because the shoulder suspension apparatus is subjected to asymmetrical forces from handedness, we predicted that its skeletal features would grow asymmetrically. We used this prediction to test our hypothesis in a natural experiment to correlate the size of the skeletal features with the forces exerted on them. We (1) measured biomechanically relevant bony features within the shoulder suspension apparatus in 101 male human specimens (62 of known handedness); and (2) modeled and analyzed the forces within the shoulder suspension apparatus from X-ray CT data. We identified eight right-handed characters and demonstrated the causal relationship between these right-handed characters and the magnitude and direction of forces acting on them. Our data suggest that the presence of the shoulder suspension apparatus in humans was a necessary precondition for human bipedality.

Assessment of the mass, length, center of mass, and principal moment of inertia of body segments in adult males of the brown anole (Anolis sagrei) and green, or carolina, anole (Anolis carolinensis).

This study provides a morphometric data set of body segments that are biomechanically relevant for locomotion in two ecomorphs of adult male anoles, namely, the trunk-ground Anolis sagrei and the trunk-crown Anolis carolinensis. For each species, 10 segments were characterized, and for each segment, length, mass, location of the center of mass, and radius of gyration were measured or calculated, respectively. The radii of gyration were computed from the moments of inertia by using the double swing pendulum method. The trunk-ground A. sagrei has relatively longer and stockier hindlimbs and forelimbs with smaller body than A. carolinensis. These differences between the two ecomorphs demonstrated a clear relationship between morphology and performance, particularly in the context of predator avoidance behavior, such as running or jumping in A. sagrei and crypsis in A. carolinensis. Our results provide new perspectives on the mechanism of adaptive radiation as the limbs of the two species appear to scale via linear factors and, therefore, may also provide explanations for the mechanism of evolutionary changes of structures within an ecological context.

Do geese fully develop brood patches? A histological analysis of lesser snow geese (Chen caerulescens caerulescens) and Ross's geese (C. rossii).

Most birds develop brood patches before incubation; epidermis and dermis in the brood patch region thicken, and the dermal connective tissue becomes increasingly vascularized and infiltrated by leukocytes. However, current dogma states that waterfowl incubate without modifications of skin within the brood patch region. The incubation periods of lesser snow geese (Chen caerulescens caerulescens; hereafter called snow geese) and Ross's geese (C. rossii) are 2-6 days shorter than those of other goose species; only females incubate. Thus, we hypothesized that such short incubation periods would require fully developed brood patches for sufficient heat transfer from incubating parents to eggs. We tested this hypothesis by analyzing the skin histology of abdominal regions of snow and Ross's geese collected at Karrak Lake, Nunavut, Canada. For female snow geese, we found that epidermis and dermis had thickened and vascularization of dermis was 14 times greater, on average, than that observed in males (n=5 pairs). Our results for Ross's geese (n=5 pairs) were more variable, wherein only one of five female Ross's geese fully developed a brood patch. Our results are consistent with three hypotheses about brood patch development and its relationship with different energetic cost-benefit relationships, resulting from differences in embryonic development and body size.

Development and evolution of the amniote integument: current landscape and future horizon.

This special issue on the development and evolution of the amniote integument begins with a discussion of the adaptations to terrestrial conditions, the acquisition of water-impermeability of the reptilian integument, and the initial formation of filamentous integumentary appendages that prepare the way towards avian flight. Recent feather fossils are reviewed, and a definition of feathers is developed. Hierarchical models are proposed for the formation of complex structures, such as feathers. Molecular signals that alter the phenotype of integumentary appendages at different levels of the hierarchy are presented. Tissue interactions and the roles of keratins in evolution are discussed and linked to their bio-mechanical properties. The role of mechanical forces on patterning is explored. Elaborate extant feather variants are introduced. The regeneration/gene mis-expression protocol for the chicken feather is established as a testable model for the study of biological structures. The adaptations of the mammalian distal limb end organs to terrestrial, arboreal and aquatic conditions are discussed. The development and cycling of hair are reviewed from a molecular perspective. These contributions reveal that the structure and function of diverse integumentary appendages are variations that are superimposed on a common theme, and that their formation is modular, hierarchical and cyclical. They further reveal that these mechanisms can be understood at the molecular level, and that an integrative and organismal approach to studying integumentary appendages is called for. We propose that future research should foster interdisciplinary approaches, pursue understanding at the cellular and molecular level, analyze interactions between the environment and genome, and recognize the contributions of variation in morphogenesis and evolution.

The role of mechanical forces on the patterning of the avian feather-bearing skin: A biomechanical analysis of the integumentary musculature in birds.

The integumentary musculature of birds consists of three distinct components. The smooth musculature comprises feather and apterial muscles, which form a continuous musculo-elastic layer within the dermis. The feather muscles, which consistently include at least erectors and depressors, interconnect contour feathers within pterylae (i.e., feather tracts) along gridlines that are oriented diagonally to the longitudinal and transverse axes of the body. The apterial muscles interconnect pterylae by attaching to the contour feathers along their peripheries. The striated musculature is composed of individual subcutaneous muscles, most of which attach to contour feathers along the caudal periphery of pterylae A new integrative functional analysis of the integumentary musculature proposes how apterial muscles stabilize the pterylae and modulate the tension of the musculo-elastic layer, and how subcutaneous muscles provide the initial stimulus for erector muscles being able to ruffle the contour feathers within pterylae. It also shows how the arrangement of the contour feathers and integumentary muscles reflects the stresses and strains that act on the avian skin. These mechanical forces are in effect not only in the adult, especially during flight, but may also be active during feather morphogenesis. The avian integument with its complex structural organization may, therefore, represent an excellent model for analyzing the nature of interactions between the environment and genetic material. The predictions of our model are testable, and our study demonstrates the relevance of integrated analyses of complex organs as mechanically coherent systems for evolutionary and developmental biology.