Oberhautchen Layer Research Articles

Microscopic structure and immunolabeling of extremely overlapped scales in some scincid, anguid, and pygopod lizards.

Skink, anguid, and pygopod lizards possess an extremely flat skin, imparting a compact and solid body and shining surface that facilitates their slider and/or fossorial movements. The present morphological study, conducted using immunohistochemistry and electron microscopy, has analyzed the microscopical morphology of extremely overlapped scales in different lizards, including species with limb reduction (scincids such as Lerista bougainvilli, Scincella lateralis, Lampropholis delicata) or legless (pygopods such as Lialis burtonis and Delma molleri and the anguid Anguis fragilis). The outer surface of the epidermis shows different micro-structures of the Oberhautchen layer containing corneous beta-proteins (CBPs) with variable immunoreactivity for these proteins. The beta-layer is relatively thick in most of these species, probably in relation to the resistance against strong mechanical forces acting on scales during the movements on harsh substrates. The scincid and anguid lizards also possess and regenerate osteoderms that reinforce scales flatness and mechanical resistance during the serpentiform or fossorial movements of these reptiles. Osteoderms are absent in pygopods. Roundish cells with a granular content are detected in the deep hinge region of scales in Lerista and Lampropholis skinks. Whether these cells may secrete substances that facilitate scale anti-friction and also determine shining of the skin surface remains to be shown.

Skin structure of the slow worm lizard Anguis fragilis (Anguidae, Sauria, Reptilia) with emphasis on the epidermal micro‐ornamentation in relation to the animal movements

AbstractSkin structure of the slow worm lizard Anguis fragilis (Anguidae, Sauria, Reptilia) with emphasis on the epidermal micro‐ornamentation in relation to the animal movements (Acta Zoologica, Stockholm). The structure of the skin and superficial micro‐ornamentation in the slow worm Anguis fragilis, a limbless lizard with a fossorial activity, was examined using histology, immunofluorescence, scanning and transmission electron microscopy. The scales, with a triangular to trapezoidal shape, are very overlapped and interlocked to form a smooth surface and are reinforced by osteoderms. The epidermis shows a thin Oberhautchen layer merged with a thicker beta‐layer that contains corneous beta‐proteins. The SEM survey detects a smooth surface made of tile‐like patterned Oberhautchen cells with irregular perimeters that form an interlocking surface. Disk‐like sensory organs of 15–20 μm diameter are observed only on the head scales, the first to sense the environment and contact the ground. Numerous Oberhautchen denticles, namely corneous thorns of about 0.2–0.3 μm, adorn the caudally directed perimeter of Oberhautchen cells in the ventral scales of the trunk and tail. This microstructure may determine gripping and increased friction with the substrate during the lateral undulating and forward movements of the slow worm. TEM observations reveal sparse short serrated protrusions of Oberhautchen cells that are largely merged with the underlying beta‐cells. Altogether, the scale surface of the slow worm efficiently suites this limbless lizard to its environment and lifestyle.

Bristles formation in adhesive pads and sensilli of the gecko Tarentola mauritanica derive from a massive accumulation of corneous material in Oberhautchen cells of the epidermis

Among lizards, geckos possess special digital scales modified as hairy-like lamellae that allow attachment to vertical substrates for the movement using adhesive nanoscale filaments called setae. The present study shows new ultrastructural details on setae formation in the gecko Tarentula mauritanica. Setae derive from the special differentiation of an epidermal layer termed Oberhauchen and can reach 30–60 µm in length. Oberhautchen cells in the adhesive pad lamellae becomes hypertrophic and rest upon 2 layers of non-corneous and pale cells instead of beta-cells like in the other scales. Only 1–2 beta-layers are formed underneath the pale layer. Setae derive from the accumulation of numerous roundish and heterogenous beta-packets with variable electron-density in Oberhautchen cells, possibly indicating a mixed protein composition. Immunofluorescence and immunogold labeling for CBPs show that beta-packets merge at the base of the growing setae forming long corneous bundles. Pale cells formed underneath the Oberhautchen layer contain small vesicles or tubules with a likely lipid content, sparse keratin filaments and ribosomes. In mature lamellae these cells merge with Oberhautchen and beta-cells forming a thin electron-paler layer located between the Oberhautchen and the thin beta-layer, a variation of the typical sequence of epidermal layers present in other scales. The formation of a softer pale layer and of a thin beta-layer likely determines a flexible corneous support for the adhesive setae. The specific molecular mechanism that stimulates the cellular changes observed during Oberhautchen hypertrophy and the alteration of the typical epidermal stratification in the pad epidermis remains unknown.

Micro-ornamentation patterns in different areas of the epidermis in the gecko Tarentola mauritanica reflect variations in the accumulation of corneous material in Oberhautchen cells.

Micro-ornamentations characterize the surface of scales in lepidosaurians and are summarized in four main patterns, i.e., spinulated, lamellated, lamellate-dentate, and honeycomb, although variations of these patterns are present in different species. Although geckos are known to possess a spinulated pattern derived from the Oberhautchen layer, also other pattern variations of the spinulated micro-ornamentation are present such as those indicated as dendritic ramification, corneous belts, and small bare patches. The present study mainly describes the variation of micro-ornamentations present in scales of different skin regions in the Mediterranean gecko Tarentula mauritanica using scannig and transmission electron microscopy. The study reports that the accumulation of corneous material in Oberhautchen cells is not homogenous in different areas of body scales and, when mature, this process gives rise to different sculpturing on the epidermal surface generating not only spinulae but also transitional zones leading to the other main patterns. It is hypothesized that spinulae formation derives from the vertical and lateral symmetric growth of tubercolate, non-overlapped scales of geckos. Sparse areas also result smooth or with serpentine-ridges likely revealing the beta-layer located underneath and merged with the Oberhautchen. The eco-functional role of this variable micro-ornamentation in the skin of lizards however remains largely speculative.

Immunolocalization of corneous proteins including a serine-tyrosine-rich beta-protein in the adhesive pads in the tokay gecko.

Adhesive pads of geckos contain many thousands of nanoscale spatulae for the adhesion and movement along vertical or inverted surfaces. Setae are composed of interlaced corneous bundles made of small cysteine-glycine-rich corneous beta proteins (CBPs, formerly indicated as beta-keratins), embedded in a matrix material composed of cytoskeletal proteins and lipids. Negatively charged intermediate filament keratins (IFKs) and positively charged CBPs likely interact within setae, aside disulphide bonds, giving rise to a flexible and resistant corneous material. Using differernt antibodies against CBPs and IFKs an updated model of the composition of setae and spatulae is presented. Immunofluorescence and ultrastructural immunogold labeling reveal that one type of neutral serine-tyrosine-rich CBP is weakly localized in the setae while it is absent from the spatula. This uncharged protein is mainly present in the thin Oberhautchen layer sustaining the setae, although with a much lower intensity with respect to the cysteine-rich CBPs. These proteins in the spatula likely originate a positively charged or neutral contact surface with the substrate but the influence of lipids and cytoskeletal proteins present in setae on the mechanism of adhesion is not known. In the spatula, protein-lipid complexes may impart the pliability for the attachment and adapt to irregular surfaces. The presence of cysteine-glycine medium rich CBPs and softer IFKs in alpha-layers sustaining the setae forms a flexible base for compliance of the setae to substrate and improved adhesion.

Gecko Adhesion in Space and Time: A Phylogenetic Perspective on the Scansorial Success Story.

An evolutionary perspective on gecko adhesion was previously hampered by a lack of an explicit phylogeny for the group and of robust comparative methods to study trait evolution, an underappreciation for the taxonomic and structural diversity of geckos, and a dearth of fossil evidence bearing directly on the origin of the scansorial apparatus. With a multigene dataset as the basis for a comprehensive gekkotan phylogeny, model-based methods have recently been employed to estimate the number of unique derivations of the adhesive system and its role in lineage diversification. Evidence points to a single basal origin of the spinulate oberhautchen layer of the epidermis, which is a necessary precursor for the subsequent elaboration of a functional adhesive mechanism in geckos. However, multiple gains and losses are implicated for the elaborated setae that are necessary for adhesion via van der Waals forces. The well-supported phylogeny of gekkotans has demonstrated that convergence and parallelism in digital design are even more prevalent than previously believed. It also permits the reexamination of previously collected morphological data in an explicitly evolutionary context. Both time-calibrated trees and recently discovered amber fossils that preserve gecko toepads suggest that a fully-functional adhesive apparatus was not only present, but also represented by diverse architectures, by the mid-Cretaceous. Further characterization and phylogenetically-informed analyses of the other components of the adhesive system (muscles, tendons, blood sinuses, etc.) will permit a more comprehensive reconstruction of the evolutionary pathway(s) by which geckos have achieved their structural and taxonomic diversity. A phylogenetic perspective can meaningfully inform functional and performance studies of gecko adhesion and locomotion and can contribute to advances in bioinspired materials.

Review: mapping proteins localized in adhesive setae of the tokay gecko and their possible influence on the mechanism of adhesion.

The digital adhesive pads that allow gecko lizards to climb vertical surfaces result from the modification of the oberhautchen layer of the epidermis in normal scales. This produces sticky filaments of 10-100μm in length, called setae that are composed of various proteins. The prevalent types, termed corneous beta proteins (CBPs), have a low molecular weight (12-20kDa) and contain a conserved central region of 34 amino acids with a beta-conformation. This determines their polymerization into long beta-filaments that aggregate into corneous beta-bundles that form the framework of setae. Previous studies showed that the prevalent CBPs in the setae of Gekko gecko are cysteine-rich and are distributed from the base to the tip of adhesive setae, called spatulae. The molecular analysis of these proteins, although the three-dimensional structure remains undetermined, indicates that most of them are charged positively and some contain aromatic amino acids. These characteristics may impede adhesion by causing the setae to stick together but may also potentiate the van der Waals interactions responsible for most of the adhesion process on hydrophobic or hydrophilic substrates. The review stresses that not only the nanostructural shape and the high number of setae present in adhesive pads but also the protein composition of setae influence the strength of adhesion to almost any type of substrate. Therefore, formulation of dry materials mimicking gecko adhesiveness should also consider the chemical nature of the polymers utilized to fabricate the future dry adhesives in order to obtain the highest performance.

Presence of a glycine-cysteine-rich beta-protein in the oberhautchen layer of snake epidermis marks the formation of the shedding layer.

The complex differentiation of snake epidermis largely depends on the variation in the production of glycine-cysteine-rich versus glycine-rich beta-proteins (beta-keratins) that are deposited on a framework of alpha-keratins. The knowledge of the amino acid sequences of beta-proteins in the snake Pantherophis guttatus has allowed the localization of a glycine-cysteine-rich beta-protein in the spinulated oberhautchen layer of the differentiating shedding complex before molting takes place. This protein decreases in the beta-layer and disappears in mesos and alpha-layers. Conversely, while the mRNA for a glycine-rich beta-protein is highly expressed in differentiating beta-cells, the immunolocalization for this protein is low in these cells. This discrepancy between expression and localization suggests that the epitope in glycine-rich beta-proteins is cleaved or modified by posttranslational processes that take place during the differentiation and maturation of the beta-layer. The present study suggests that among the numerous beta-proteins coded in the snake genome to produce epidermal layers with different textures, the glycine-cysteine-rich beta-protein marks the shedding complex formed between alpha- and beta-layers that allows for molting while its disappearance between the beta- and alpha-layers (mesos region for scale growth) is connected to the formation of the alpha-layers.

Immunolocalization of Keratin‐Associated Beta‐Proteins (Beta‐Keratins) in Pad Lamellae of Geckos Suggest that Glycine–Cysteine‐Rich Proteins Contribute to Their Flexibility and Adhesiveness

The epidermis of digital pads in geckos comprises superficial microornamentation from the oberhautchen layer that form long setae allowing these lizards to climb vertical surfaces. The beta-layer is reduced in pad lamellae but persists up to the apical free margin. Setae are made of different proteins including keratin-associated beta-proteins, formerly indicated as beta-keratins. In order to identify specific setal proteins the present ultrastructural study on geckos pad lamellae analyzes the immunolocalization of three beta-proteins previously found in the epidermis and adhesive setae of the green anolis. A protein rich in glycine but poor in cysteine (HgG5-like) is absent or masked in gecko pad lamellae. Another protein rich in glycine and cysteine (HgGC3-like) is weakly present in setae, oberhautchen and beta-layer. A glycine and cysteine medium rich beta-protein (HgGC10-like) is present in the lower part of the beta-layer but is absent in the oberhautchen, setae, and mesos layer. The latter two proteins may form intermolecular bonds that contribute to the flexibility of the corneous material sustaining the setae. The pliable alpha-layer present beneath the thin beta-layer and in the hinge region of the pad lamellae also contains HgGC10-like proteins. Based on the possibility that some HgGC3-like or other cys-rich beta-proteins are charged in the setae it is suggested that their charges influence the mechanism of adhesion increasing the induction of dipoles on the substrate and enhancing attractive van der Waals forces.

The Epidermis of Scales in Gecko Lizards Contains Multiple Forms of β-Keratins Including Basic Glycine-Proline-Serine-Rich Proteins

The epidermis of scales of gecko lizards comprises alpha- and beta-keratins. Using bidimensional electrophoresis and immunoblotting, we have characterized keratins of corneous layers of scales in geckos, especially beta-keratins in digit pad lamellae. In the latter, the formation of thin bristles (setae) allow for the adhesion and climbing vertical or inverted surfaces. alpha-Keratins of 55-66 kDa remain in the acidic and neutral range of pI, while beta-keratins of 13-18 kDa show a broader variation of pI (4-10). Some protein spots for beta-keratins correspond to previously sequenced, basic glycine-proline-serine-rich beta-keratins of 169-191 amino acids. The predicted secondary structure shows that a large part of the molecule has a random-coiled conformation, small alpha helix regions, and a central region with 2-3 strands (beta-folding). The latter, termed core-box, shows homology with feather-scale-claw keratins of birds and is involved in the formation of beta-keratin filaments. Immunolocalization of beta-keratins indicates that these proteins are mainly present in the beta-layer and oberhautchen layer, including setae. The sequenced proteins of setae form bundles of keratins that determine their elongation. This process resembles that of feather-keratin on the elongation of barbule cells in feathers. It is suggested that small proteins rich in glycine, serine, and proline evolved in reptiles and birds to reinforce the mechanical resistance of the cytokeratin cytoskeleton initially present in the epidermis of scales and feathers.

Differentiation of snake epidermis, with emphasis on the shedding layer

Little is known about specific proteins involved in keratinization of the epidermis of snakes. The presence of histidine-rich molecules, sulfur, keratins, loricrin, transglutaminase, and isopeptide-bonds have been studied by ultrastructural autoradiography, X-ray microanalysis, and immunohistochemistry in the epidermis of snakes. Shedding takes place along a shedding complex, which is composed of two layers, the clear and the oberhautchen layers. The remaining epidermis comprises different layers, some of which contain beta-keratins and others alpha-keratins. Weak loricrin, transglutaminase, and sometimes also iso-peptide-bond immunoreactivities are seen in some cells, lacunar cells, of the alpha-layer. Tritiated histidine is mainly incorporated in the shedding complex, especially in dense beta-keratin filaments in cells of the oberhautchen layer and to a small amount in cells of the clear layer. This suggests the presence of histidine-rich, matrix proteins among beta-keratin bundles. The latter contain sulfur and are weakly immunolabeled for beta-keratin at the beginning of differentiation of oberhautchen cells. After merging with beta cells, the dense beta-keratin filaments of oberhautchen cells become immunopositive for beta-keratin. The uptake of histidine decreases in beta cells, where little dense matrix material is present, while pale beta-keratin filaments increase. During maturation, little histidine labeling remains in electron-dense areas of the beta layer and in those of oberhautchen spinulae. Some roundish dense granules of oberhautchen cells rich in sulfur are negative to antibodies for alpha-keratin, beta-keratin, and loricrin. The granules eventually merge with beta-keratin, and probably contribute to the formation of the resistant matrix of oberhautchen cells. In conclusion, beta-keratin, histidine-rich, and sulfur-rich proteins contribute to form snake microornamentations.

Synthesis of interkeratin matrix in differentiating lizard epidermis: an ultrastructural autoradiographic study after injection of tritiated proline and histidine.

During epidermal differentiation in mammals, keratins and keratin-associated matrix proteins rich in histidine are synthesized to produce a corneous layer. Little is known about interkeratin proteins in nonmammalian vertebrates, especially in reptiles. Using ultrastructural autoradiography after injection of tritiated proline or histidine, the cytological process of synthesis of beta-keratin and interkeratin material was studied during differentiation of the epidermis of lizards. Proline is mainly incorporated in newly synthesized beta-keratin in beta-cells, and less in oberhautchen cells. Labeling is mainly seen among ribosomes within 30 min postinjection and appears in beta-keratin packets or long filaments 1-3 h later. Beta-keratin appears as an electron-pale matrix material that completely replaces alpha-keratin filaments in cells of the beta-layer. Tritiated histidine is mainly incorporated into keratohyalin-like granules of the clear layer, in dense keratin bundles of the oberhautchen layer, and also in dense keratin filaments of the alpha and lacunar layer. The detailed ultrastructural study shows that histidine-labeling is localized over a dense amorphous material associated with keratin filaments or in keratohyalin-like granules. Large keratohyalin-like granules take up labeled material at 5-22 h postinjection of tritiated histidine. This suggests that histidine is utilized for the synthesis of keratins and keratin-associated matrix material in alpha-keratinizing cells and in oberhautchen cells. As oberhautchen cells fuse with subjacent beta-cells to form a syncytium, two changes occur : incorporation of tritiated histidine, but uptake of proline increases. The incorporation of tritiated histidine in oberhautchen cells lowers after merging with cells of the beta-layer, whereas instead proline uptake increases. In beta-cells histidine-labeling is lower and randomly distributed over the cytoplasm and beta-keratin filaments. Thus, change in histidine uptake somehow indicates the transition from alpha- to beta-keratogenesis. This study indicates that a functional stratum corneum in the epidermis of amniotes originates only after the association of matrix and corneous cell envelope proteins with the original keratin scaffold of keratinocytes.

Ultrastructural autoradiographic and immunocytochemical analysis of setae formation and keratinization in the digital pads of the gecko Hemidactylus turcicus (Gekkonidae, Reptilia)

The modified subdigital scales of some lizards allow them to climb vertical surfaces. This is due to the action of millions of tiny setae present in the digital pads. Setae are mainly composed of beta-keratin which may have some modality of aggregation similar to that of barbs and barbules of feathers. Keratins and associated proteins are involved in the organization of setae. The formation of setae in the climbing pad lamellae of the gecko Hemidactylus turcicus has been analyzed under the electron microscope after injection of tritiated histidine and immunocytochemistry for a chick scale beta-keratin. Setae are made up of dense and pale filaments, both oriented along the longer axis of setae. Beta-keratin is present in the oberhautchen layer and in the growing setae which are highly modified oberhautchen cells. Most of the immunolabeling concentrated in the central part of setae. This cross-reactivity suggests that some epitopes in chick beta-keratin are also present in gecko setae. Four hours after injection of tritiated histidine, the labeling is localized over setae, in particular in the dense filaments and less in the pale filaments. Some labeling is also seen in the keratinaceous material present in the cytoplasm of clear cells, which are believed to mold setae. The present observations suggest that both beta-keratin and denser matrix proteins, possibly incorporating histidine, are packed into growing setae. These proteins may be mixed to form pale and dense filaments oriented along the longer axis of setae, a pattern resembling that of barb and barbule cells of feathers. The role of matrix material in the orientation of the deposited beta-keratin during setal outgrowth is discussed with the problem of barb and barbule differentiation in avian feathers.

Epidermal differentiation during ontogeny and after hatching in the snake Liasis fuscus (Pythonidae, Serpentes, Reptilia), with emphasis on the formation of the shedding complex.

Differentiation and localization of keratin in the epidermis during embryonic development and up to 3 months posthatching in the Australian water python, Liasis fuscus, was studied by ultrastructural and immunocytochemical methods. Scales arise from dome-like folds in the skin that produce tightly imbricating scales. The dermis of these scales is completely differentiated before any epidermal differentiation begins, with a loose dermis made of mesenchymal cells beneath the differentiating outer scale surface. At this stage (33) the embryo is still unpigmented and two layers of suprabasal cells contain abundant glycogen. At Stage 34 (beginning of pigmentation) the first layers of cells beneath the bilayered periderm (presumptive clear and oberhautchen layers) have not yet formed a shedding complex, within which prehatching shedding takes place. At Stage 35 the shedding complex, consisting of the clear and oberhautchen layers, is discernible. The clear layer contains a fine fibrous network that faces the underlying oberhautchen, where the spinulae initially contain a core of fibrous material and small beta-keratin packets. Differentiation continues at Stage 36 when the beta-layer forms and beta-keratin packets are deposited both on the fibrous core of the oberhautchen and within beta-cells. Mesos cells are produced from the germinal layer but remain undifferentiated. At Stage 37, before hatching, the beta-layer is compact, the mesos layer contains mesos granules, and cells of the alpha-layer are present but are not yet keratinized. They are still only partially differentiated a few hours after hatching, when a new shedding complex is forming underneath. Using antibodies against chick scale beta-keratin resolved at high magnification with immunofluorescent or immunogold conjugates, we offer the first molecular confirmation that in snakes only the oberhautchen component of the shedding complex and the underlying beta cells contain beta-keratin. Initially, there is little immunoreactivity in the small beta-packets of the oberhautchen, but it increases after fusion with the underlying cells to produce the syncytial beta layer. The beta-keratin packets coalesce with the tonofilaments, including those attached to desmosomes, which rapidly disappear in both oberhautchen and beta-cells as differentiation progresses. The labeling is low to absent in forming mesos-cells beneath the beta-layer. This study further supports the hypothesis that the shedding complex in lepidosaurian reptiles evolved after there was a segregation between alpha-keratogenic cells from beta-keratogenic cells during epidermal renewal.

Histidine uptake in the epidermis of lizards and snakes in relation to the formation of the shedding complex.

Mammalian epidermis utilizes histidine-rich proteins (filaggrins) to aggregate keratin filaments and form the stratum corneum. Little is known about the involvement of histidine-rich proteins during reptilian keratinization. The formation of the shedding complex in the epidermis of snakes and lizards, made of the clear and the oberhautchen layers, determines the cyclical epidermal sloughing. Differently from snakes, keratohyalin-like granules are present in the clear layer of lizards. The uptake of tritiated histidine into the epidermis of two lizards and one snake has been studied by autoradiography in sections at progressive post-injection periods. At 40 min and 1 hr post-injection keratohyalin-like granules were not or poorly labeled. At 3-22 hr post-injection most of the labeling was present over suprabasal cells destined to form the shedding complex, in keratohyalin-like granules of the clear layer, and in the forming a-layer but was low in the forming b-layer, and in superficial keratinized layers. The analysis of the shedding complex in the pad lamellae (a specialized scale used for climbing) of a gecko showed that the setae and the cytoplasm of clear cells among them are main sites of histidine uptake at 4 hr post-injection. In the snake most of the labeling at 4 hr post-injection was localized in the shedding complex along the boundary between the clear and oberhautchen layers. The present study suggests that, in the epidermis of lepidosaurian reptiles, the synthesis of a histidine-rich protein is involved in the formation of the shedding layer and, as in mammals, in a-keratinization.

Epidermal differentiation in the developing scales of embryos of the Australian scincid lizard Lampropholis guichenoti.

Formation of the first epidermal layers in the embryonic scales of the lizard Lampropholis guichenoti was studied by optical and electron microscopy. Morphogenesis of embryonic scales is similar to the general process in lizards, with well-developed overlapping scales being differentiated before hatching. The narrow outer peridermis is torn and partially lost during scale morphogenesis. A second layer, probably homologous to the inner peridermis of other lizard species, but specialized to produce lipid-like material, develops beneath the outer peridermis. Two or three lipogenic layers of this type develop in the forming outer surface of scales near to the hinge region. These layers form a structure here termed "sebaceous-like secretory cells." These cells secrete lipid-like material into the interscale space so that the whole epidermis is eventually coated with it. This lipid-like material may help to reduce friction and to reduce accumulation of dirt between adjacent extremely overlapping scales. At the end of their differentiation, the modified inner periderm turns into extremely thin cornified cells. The layer beneath the inner peridermis is granulated due to the accumulation of keratohyalin-like granules, and forms a shedding complex with the oberhautchen, which develops beneath. Typically tilted spinulae of the oberhautchen are formed by the aggregation of tonofilaments into characteristically pointed cytoplasmic outgrowths. Initially, there is little accumulation of beta-keratin packets in these cells. During differentiation, the oberhautchen layer merges with cells of the beta-keratin layer produced underneath, so that a typical syncytial beta-keratin layer is eventually formed before hatching. Between one-fourth distal and the scale tip, the dermis under epidermal cells is scarce or absent so that the mature scale tip is made of a solid rod of beta-keratinized cells. At the time of hatching, differentiation of a mesos layer is well advanced, and the epidermal histology of scales corresponds to Stage 5 of an adult shedding cycle. The present study confirms that the embryonic sequence of epidermal stratification observed in other species is basically maintained in L. guichenoti.

Keratohyalin-like granules in embryonic and regenerating epidermis of lizards and Sphenodon punctatus (Reptilia, Lepidosauria)

AbstractEpidermal shedding in lepidosaurians is determined by the formation of an intraepidermal shedding complex, made of an upper clear layer which interdigitates with a lower oberhautchen layer. The latter produces species-specific microornamentations. The ultrastructure of embryonic and regenerating epidermis in seven species of lizards representing five different squamate families (Gekkonidae, Agamidae, Lacertidae, Iguanidae and Scincidae), and on the regenerating caudal epidermis of the tuatara (Sphenodon punctatus, suborder Sphenodontia), has shown a broad variation in dimension and morphology of keratohyalin-like granules present in the clear layer of the shedding complex. Where the oberhautchen ornamentations are well developed, like in the setae of the climbing pads, keratohyalin-like granules appear more irregular, larger or more numerous. The present study suggests that keratohyalin-like granules might be involved in the formation of a fibrous network around the forming oberhautchen microornamentations.

Morphogenesis of the digital pad lamellae in the embryo of the lizard Anolis lineatopus

The present study in the embryo of the lizard Anolis lineatopus describes the modality of cell proliferation responsible for the morphogenesis of the digital pad lamellae and of the epidermal stratification. After tritiated thymidine and 5‐bromodeoxy‐uridine administration, autoradiographic and immunocytochemical methods have been used. The lamellae originate as long, slightly slanted, undulations of the epidermis of fingers and toes. At an early stage, the epidermis consists of an outer periderm and a basal layer. Cell hypertrophy, and the prevalent cell proliferation in the longer side of the undulation with respect to the shorter side, generate the surface of the outer lamella. Under the peridermis, a shedding complex, composed by clear and oberhautchen layers, is formed and later determines the first intraepidermal shed. The first subperidermal layer derived from the basal layer is a clear layer and the first shed epidermis in the embryo is represented by periderm and clear layer. The heavily granulated clear layer in Anolis lineatopus represents the first epidermal layer produced in the embryonic epidermis, and is connected with the process of shedding. The spinulae of the underlying oberhautchen in the outer scale surface become long setae which grow toward the upper clear layer. Under the shedding complex a β‐layer is produced. Autoradiographical study shows that the radioactivity stays in the basal layer for about 4 days before cells move to upper layers. At 6–8 days post‐injection labelled cells are visible in the differentiated clear, oberhautchen and β‐layers. Under the β‐layer differentiating mesos cells are visible before the embryo hatches.

Microstructure, ontogeny, and evolution of scale surfaces in xenosaurid lizards.

Both scanning electron and light microscopy were used to examine the epidermal structure of scales taken from several ontogenetic stages of Xenosaurus grandis and Shinisaurus crocodilurus. In addition, scales from all xenosaurid species were examined by scanning electron microscopy to determine scale surface variation among genera, species, and subspecies. A varied and phylogenetically informative morphology characterizes the scale surfaces of xenosaurid lizards. Scale surface morphology is conservative among the species and subspecies of Xenosaurus, but is more variable between the two xenosaurid genera. Their scale surfaces are characterized by folds in the oberhautchen, beta, mesos, and alpha epidermal layers, forming polygonal ridges of a type previously described for the Iguania. The three species of Xenosaurus possess lenticular scale organs, whereas Shinisaurus has scale organs with spikes (bristles). The spikes of Shinisaurus are formed by the beta and oberhautchen layers, with the alpha layer forming a dome-shaped cap over a dermal papilla. Shinisaurus crocodilurus exhibits a dramatic ontogenetic change in scale surface morphology, that is here reported for the first time in any lizard. © 1993 Wiley-Liss, Inc.

Scanning electron microscopy of scales from different body regions of three lizard species

The Oberhautchen of scales from the dorsal, parietal, and ventral regions of Sceloporus occidentalis (Iguanidae), Gerrhonotus multicarinatus (Anguinidae), and Anniella pulchra (Anniellidae) were examined with a scanning electron microscope. At low magnification, all scales of S. occidentalis exhibit well-defined outlines of cells belonging to the Oberhautchen layer and the previously overlying clear layer. The dorsal and parietal cells of this species exhibit a minutely dentate Oberhautchen that forms tooth-like spinules 0.2 to 0.5 μ long and arranged in irregular rows. Minute pits 0.1 to 0.3 μ in diameter characterize the Oberhautchen of a ventral scale. Cell outlines are not evident on the scales of G. multicarinatus. The Oberhautchen of dorsal and parietal scales of this species is prominently laminated. Laminae are less prominent on scales of the lateral fold, and no intrinsic surface structure is evident on a ventral scale. In contrast, the fossorial anguinomorph Anniella pulchra exhibits Oberhautchen surfaces with practically no intrinsic microornamentation. However, what appear to be outlines of Oberhautchen cells are visible on the dorsal and ventral scales. These observations suggest that modifications of Oberhautchen microornamentation may have evolved to reduce friction with the substrate or other scales. The lack of pronounced microornamentation of the Oberhautchen on some body scales may indicate that a complex interdigitation between clear layer and Oberhautchen cells is not essential to the sloughing process.