Spermatogenesis and ultrastructure of the spermatozoa of Seison nebaliae (Syndermata)

 The spermatozoa of Seison nebaliae are characterized by an elongated sperm body, a filiform nucleus, and an anteriorly inserting external cilium with a 9×2+2 axoneme pattern. In the sperm body a frontal, middle, and hind region can be distinguished. The frontal region contains an acrosomal vesicle, a perforatorium, a basal body, and a pair of apical dense bodies; an accessory centriole is absent. The middle region is characterized by several so-called filamental plates. One large mitochondrion and one pair of accessory tubular structures are located in the middle and hind region. The hind region also contains two rows of dense bodies. Accessory tubular structures and filamental plates are autapomorphies of S. nebaliae. The shared appearance of the dense bodies in spermatozoa of species of the taxa Seison and the Acanthocephala founds their sister-group relationship, while the anterior insertion of the cilium in the spermatozoa of these taxa and in the Rotifera confirms the monophylum Syndermata Ahlrichs, 1995. Accepted: 5 August 1998     

Musculature of Seison nebaliae Grube, 1861 and Paraseison annulatus (Claus, 1876) revealed with CLSM: a comparative study of the gnathiferan key taxon Seisonacea (Rotifera)

The somatic muscular systems of two species of Seisonacea (Rotifera), Seison nebaliae and Paraseison annulatus, are described using fluorescently labelled phalloidin in combination with confocal laser scanning microscopy. Their overall muscular arrangement is similar and consists of segmentally organised longitudinal fibres that extend the length of the body and are surrounded by semi-circular (= incomplete) bands. However, differences in the musculature between the two species are present and possibly reflect specific adaptations in feeding strategy and locomotion related to the occupation of individual niches on their host, the leptostracan crustacean N. bipes. For example, S. nebaliae has semi-circular muscles in the head region only, while P. annulatus possesses incomplete circular muscles also in the trunk region; furthermore, there are also differences in the arrangements and number of longitudinal muscles. The muscular systems of all rotifer species examined so far are compared in order to establish the ground pattern of the last putative ancestor as well as to seek for traits of systematic importance. Results from both species corroborate earlier hypotheses on the arrangement of muscles in the putative common ancestor of Rotifera, which suggested an orthogonal arrangement consisting of a series of probably continuous (not segmental) inner longitudinal muscles, surrounded by semi-circular fibres, ventrally opened. However, significant morphological and ecological variations among taxa investigated so far show that a consistent correlation between muscular traits and specific ecological features and/or phylogeny is still far from being clear. Hence, musculature of additional taxa, representing the systematic width and occupying a diverse range of habitats, should be investigated.     

Ultrastructure of the protonephridia of Seison annulatus (Rotifera)

Summary Each of the two protonephridial systems of Seison annulatus consists of three sections which are separated by cell borders with septate junctions: (a) a terminal syncytium with eight terminal organs and a capillary canal, (b) a canal syncytium which is divided into a multiciliary canal region and a main canal region, and (c) a nephroporus cell. The terminal syncytium is branched and linked twice to the canal syncytium. The supporting structure of each filtration barrier is a hollow cylinder which is perforated by pores and lacks microvilli (pillars). A protonephridial spine is situated in the multiciliary canal region and stabilizes the neck region. The ored, hollow cylinder and the protonephridial spine are new characteristics for the Rotifera.     

Spermiogenesis in Seison nebaliae (Rotifera, Seisonidea): further evidence of a rotifer-acanthocephalan relationship

The spermatozoa of Seison nebaliae are filiform cells about 70 mum long with a diameter of 0.6 mum. They have a slightly enlarged head, 2.5 mum long, followed by a long cell body. The flagellum starts from the head, and runs parallel to the cell body, contained in a groove along it. The head contains an acrosome, two large, paired para-acrosomal bodies, the basal body of the flagellum and the anterior thin extremity of the nucleus. The cell body contains the main portion of the nucleus, a single mitochondrion located in its distal portion, and many accessory bodies with different shapes. The flagellum has a 9 + 2 axoneme. The study of spermiogenesis shows the Golgian origin of the acrosome and the para-acrosomal bodies and reveals some peculiarities: a folding of the perinuclear cisterna is present between the proacrosome and the basal body of the flagellum in early spermatids and the flagellum runs in a canal inside the spermatid cytoplasm. The basal body migrates anteriorly. These characters are shared partly by the Rotifera Monogononta and, to a large extent, by the Acanthocephala studied so far. Many details of the spermiogenetic process are identical to those of Acanthocephala, thus suggesting that the processes in the two taxa are homologous.     

Comparative ultrastructure of the epidermal ciliary rootlets and associated structures in species of the Nemertodermatida and Acoela (Plathelminthes)

 The fine morphology of epidermal ciliary structures in four species of the Nemertodermatida and four species of the Acoela was studied, with emphasis on Meara stichopi (Nemertodermatida). The cilium of M. stichopi has a distal shelf and is proximally separated from the basal body by a cup-shaped structure. The bottom of the cup consists of a bilayered dense plate, or basal plate. The basal body consists of peripheral microtubule doublets continuous with those of the cilium. In the upper part of the basal body, the doublets are set at an angle and are anchored to the enclosing cell membrane by Y-shaped structures. The lower part of the basal body tapers eventually. The striated main rootlet arises on the anterior face of the basal body, initially like a flattened strap, and continues along the basal body shaped as a tube which further down becomes solid. The hour-glass-shaped posterior rootlet arises on the posterior face of the basal body. Contrary to the main rootlet, the striations in the proximal part of the posterior rootlet run parallel to the microtubule doublets of the basal body. A pair of microtubule bundles lead from the posterior rootlet to the two main rootlets in the hind ciliary row, and follow these to their lower tip. In the other species of the Nemertodermatida studied, the structure of the ciliary basal body and the ciliary rootlets is similar to that of M. stichopi. Structural differences in the species of the Acoela are that the lowermost end of the basal body is narrow and bent forwards, the proximal part of the main rootlet is trough-shaped, the main rootlet is accompanied by a pair of lateral rootlets and the posterior rootlet with associated microtubule bundles is thin. The epidermal ciliary structures in species of the Nemertodermatida and Acoela have a number of shared characters which are unique within the Plathelminthes. However, almost all of these characters are found in Xenoturbella bocki (Xenoturbellida), and some even in species of other ”phyla” of the ”lower” Metazoa. Hence, these characters cannot be considered apomorphic for the Acoelomorpha. A character seemingly present only in species of the Nemertodermatida and Acoela is the bilayered dense plate. This feature might represent an autapomorphic character state for the Acoelomorpha. Accepted: 7 March 1997     

Epidermal stem cells: the cradle of epidermal determination, differentiation and wound healing

The field of epidermal stem cells has dramatically advanced in the last decade, leading to a better understanding of the molecular factors, signalling pathways and cellular events that identify and characterize stem cells, thus revealing their immense potential for therapeutic use. Furthermore, multipotent epidermal stem cells present the major advantage of easy accessibility with the discovery of their specific location within the bulge of the hair follicle. This review focuses on the most recent findings on epidermal stem cells, and their potential role in initial epidermal commitment, differentiation and wound healing processes in the skin.     

Epidermal keratinization in the salamander and a comparison with other amphibia

In a variety of amphibians examined the stratum corneum was one cell in depth, although in Xenopus it was up to three cells deep. The flattened horny cells were closely fused together along their lateral membranes to form a continuous sheet. Disulphide bonds of keratin were most concentrated in the peripheral cytoplasm, but the interiors of the cornified cells were sufficiently well keratinized to prevent more than slight enzymatic cytolysis of the normal cell components. Characteristically large, weakly stainable, non-shrunken nuclear remnants were found in the salamander and frog horny layers, but the clawed toad had small pyknotic (parakeratotic) nuclei. The mature amphibian keratinocytes contained free fats, bound phospholipids, calcium and sulphydryl groups, together with acid phosphatase and non-specific esterase. Cornification appears to begin by a process of separate individual cell keratinization and lateral membranes of neighbouring cells only later become fused together. This differs from the process in higher vertebrates in which the cells undergoing keratinization form a uniform transitional layer in the epidermis. In the amphibian epidermis neighbouring cells occur in different stages of keratinization.     

Epidermal growth factor-induced truncation of the epidermal growth factor receptor

NIH-3T3 cells expressing the human epidermal growth factor (EGF) receptor were used in experiments to determine the fate of the EGF receptor in cells continuously exposed to EGF. EGF receptor was immunoprecipitated from cells labeled for 12 h with [35S] methionine in the absence or presence of 10 nM EGF. As expected, a single Mr = 170,000 polypeptide representing the mature EGF receptor was immune-precipitated from control cells. Surprisingly, immune precipitates from EGF-treated cells contained a prominent Mr = 125,000 receptor species, in addition to the Mr = 170,000 mature receptor. The Mr = 125,000 species was shown to be derived from the Mr = 170,000 form by pulse-chase experiments, in which the Mr = 170,000 receptor chased into the Mr = 125,000 form when EGF was included during the chase and by partial proteolysis. Both proteins became extensively phosphorylated on tyrosine residues in immune precipitate kinase assays. Treatment of immune precipitates with endoglycosidase F changed the apparent molecular weight of the Mr = 170,000 receptor to Mr = 130,000 and of the Mr = 125,000 form to Mr = 105,000, indicating that the appearance of the Mr = 125,000 protein was probably due to proteolysis. Antibody against the carboxyl terminus of the mature EGF receptor recognized the Mr = 125,000 protein, whereas antibody against the amino terminus did not. Incubation of cells with leupeptin prior to and during EGF addition inhibited processing to the Mr = 125,000 species. Methylamine and low temperature also inhibited the EGF-induced processing to the Mr = 125,000 form. These data suggest a possible role for proteolysis of the EGF receptor in receptor function.     

Isolation, characterization and comparison of mammalian epidermal prekeratins

1. The lower living layers of mammalian epidermis contain a cytoplasmic tonofilament protein, prekeratin, believed to be the precursor of the keratin which is found in the outer dead cell layer or stratum corneum. 2. Prekeratin is distinguished by its property of being extractable from epidermis homogenized in the presence of citric acid trisodium citrate buffer pH 2.65. 3. In the present study we have compared the epidermal prekeratins from ten mammalian species and have shown them to be of similar amino acid composition. 4. Conditions have been established for studying the immunology of these insoluble proteins and examination of their immunological properties has shown that they are similar to one another but that their antigenic determinants are different from those of callus keratin. 5. The SDS polyacrylamide gel electrophoretic patterns of these proteins differ widely and we have also demonstrated anatomical site variation by this method.     

Epidermal ciliary ultrastructure of adult and larval sipunculids (Sipunculida)

The ultrastructure of the ciliary apparatus of multiciliated epidermal cells in larval and adult sipunculids is described and the phylogenetic implications discussed. The pelagosphera of Apionsoma misakianum has a dense cover of epidermal cilia on the head region. The cilia have a long, narrow distal part and two long ciliary rootlets, one rostrally and one vertically orientated. The adult Phascolion strombus has cilia on the nuchal organ and on the oral side of the tentacles. These cilia have a narrow distal part as in the A. misakianum larva, but the ciliary rootlets have a different structure. The first rootlet on the anterior face of the basal body is very short and small. The second, vertically orientated rootlet is long and relatively thick. The two ciliary rootlets present in the larval A. misakianum are similar to the basal metazoan type of ciliary apparatus of epidermal multiciliated cells and thus likely represent the plesiomorphic state. The minute first rootlet in the adult P. strombus is viewed as a consequence of a secondary reduction. No possible synapomorphic character with the phylogenetically troublesome Xenoturbella was found.     

Morphology and evolution of the nervous system in Gnathostomulida (Gnathifera,Spiralia)

Within Spiralia, Gnathifera may represent the deepest branching lineage comprising the jaw worms Gnathostomulida and their sister group Micrognathozoa + Syndermata. Yet, very few nervous system studies have been conducted on this lineage of microscopic, jaw-bearing worms, limiting our understanding of the evolution of this organ system in Spiralia. The nervous system of representatives from all major groups of Gnathostomulida was here mapped using confocal laser scanning microscopy and immunohistochemistry. Their intra-epidermal, unsegmented nervous systems comprise an anterior brain and three to five ventral and two to four dorsal longitudinal nerves, connected by few transverse commissures. Neurites of the stomatogastric nervous system were found lining the pharynx and connecting to a prominent buccal ganglion. Supposedly, sensory ciliated cells in the pharynx and the gut were documented for the first time. Based on these morphological results, primary homologies of neural structures in Gnathostomulida and other Gnathifera were hypothesized and thereafter tested using parsimony. This first neurophylogeny of Gnathostomulida resulted in a topology congruent with molecular data, supporting the monophyly of Bursovaginoidea, Conophoralia, and Scleroperalia. From this topology, the evolution of the gnathostomulid nervous system was reconstructed. It suggests a specialization and diversification of cords and serotonin-like immunoreactive cell patterns from a plesiomorphic neuroarchitecture of three unsegmented nerve cords and a compact anterior brain and buccal ganglion. These plesiomorphic states resemble the nervous system of Micrognathozoa, and possibly the ancestral states of Spiralia.     

Epidermal growth factor and the onset of epithelial epidermal wound healing.

At 10 days in ovo the embryonic chick epidermis acquires the ability to spread as a cohesive epithelial sheet when wounded. A tissue culture system has been constructed that supports epidermal cell outgrowth consistent with epidermal behaviour in vivo and permits experimental manipulation of the isolated tissue with growth factors and other hormones. This culture system consists of embryonic chick epidermis isolated at days 8, 10, and 12 of development, serum-free, chemically-defined culture medium, and the inner surface of the vitelline membrane of the hen's egg as the culture substratum. At 8 days the cellular outgrowth is mesenchymal in the absence of exogenous EGF. The 8 day tissues responds to added EGF by exhibiting precocious epithelial outgrowth. The results suggest that sensitivity to EGF or EGF-like growth factors is part of the mechanism underlying the developmental onset of epidermal wound healing in skin. The epidermal origin of the outgrowth is determined by antibody staining for specific cytokeratins. The epithelial character of the outgrowth is determined by visualizing actin microfilament distribution. The normal epithelial outgrowth shows apical/basal polarization of the sheet except at the edge. From 10 days on, the isolated epidermis exhibits epithelial outgrowth from explants in culture in the absence of exogenous EGF, suggesting endogenous production of an EGF-like factor. Glucocorticoid and mineralocorticoid hormones both produce a reduced amount of epithelial outgrowth. This retardation of the early outgrowth by glucocorticoids and mineralocorticoids could result from a reduced ability of the cut edge of the epidermis to 'disorganize' and assume the unpolarized migratory form required for rapid epidermal wound healing.     

Epidermal and dermal integumentary structures of ankylosaurian dinosaurs

Ankylosaurian dinosaurs are most notable for their abundant and morphologically diverse osteoderms, which would have given them a spiky appearance in life. Isolated osteoderms are relatively common and provide important information about the structure of the ankylosaur dermis, but fossilized impressions of the soft‐tissue epidermis of ankylosaurs are rare. Nevertheless, well‐preserved integument exists on several ankylosaur fossils that shows osteoderms were covered by a single epidermal scale, but one or many millimeter‐sized ossicles may be present under polygonal, basement epidermal scales. Evidence for the taxonomic utility of ankylosaurid epidermal scale architecture is presented for the first time. This study builds on previous osteological work that argues for a greater diversity of ankylosaurids in the Dinosaur Park Formation of Alberta than has been traditionally recognized and adds to the hypothesis that epidermal skin impressions are taxonomically relevant across diverse dinosaur clades. J. Morphol. 275:39–50, 2014. © 2013 Wiley Periodicals, Inc.     

Further structures in the jaw apparatus of Limnognathia maerski (Micrognathozoa), with notes on the phylogeny of the Gnathifera

The jaws of Limnognathia maerski, Micrognathozoa, were investigated with light- and scanning electron microscopy. The study yielded several new structures and sclerites, including the ventral part of main jaw, the pharyngeal lamellae, the manus, the dorsal and ventral fibularium teeth, and a reinterpretation of the fibularium compartmentalization. Furthermore, it was shown that several jaw elements are composed of densely packed rods. Comparison with Rotifera and Gnathostomulida suggested that the micrognathozoan main jaw is homologous with the rotifer incus and the gnathostomulid articularium and that the pseudophalangids (the ventral jaws) and their associated sclerites correspond to the rotifer mallei. These results imply that Micrognathozoa is more closely related to Rotifera than to Gnathostomulida.     

Anatomy and ultrastructure of epidermal cells in the haustorium of a parasitic flowering plant,Cuscuta japonica, during attachment to the host

Changes of epidermal cells in the haustorium of the parasiticCuscuta japonica during its attachment to the host plantimpatiens balsamina were studied with light and electron microscopy. In the transverse sections of dodder stems not in contact with the host, epidermal cells had rounded outlines. However, when haustorial initials developed in the cortex of the parasite stem at the contact site, the epidermal cells had more dense cytoplasm and conspicuous nuclei than before, and their outline was flat in the longitudinal section. As meristem cells developed from those initials, the epidermal cells became more elongated. When the haustorium was fully matured, the apical tips of the elongated epidermal cells at the contact site branched like toes, producing numerous projections via cell wall invaginations. This event caused spaces to form between the projections; coincidently, the surface area of the apical ends of the epidermal cells increased. The dense cytoplasm at those projections contained prominent nuclei and abundant other organelles, suggesting a active metabolism. Osmiophilic particles, releasing into the cell walls from the cytoplasm, were though to be associated with the loosening and elongating of the epidermal cell walls. Dense and homogeneous materials were secreted within the spaces between the projections. These materials could play an important role in cementing the haustorium onto the surface of the host organ.     

Epidermal growth factor induces rapid, reversible aggregation of the purified epidermal growth factor receptor

Epidermal growth factor (EGF) receptor from A-431 cells was purified by affinity chromatography with monoclonal anti-receptor antibodies. The purified radiolabeled receptor was incubated with EGF and then analyzed by gel electrophoresis under nondenaturing conditions. In these gels, the EGF receptor migrates in two forms: a fast-migrating (low) form and an EGF-induced slow-migrating (high) form. On the basis of the various control and calibration experiments described, it is concluded that the low form represents the monomeric 170-kilodalton EGF receptor and the high form represents an EGF receptor dimer. The binding of EGF causes a rapid, temperature-sensitive dimerization of the EGF receptor. Receptor dimerization is fully reversible and involves saturable, noncovalent interactions that are stable at neutral pH and in nonionic detergents. Both the monomeric and dimeric forms of the receptor bind EGF and undergo self-phosphorylation. The dimeric form of the receptor may possess higher ligand binding affinity, and it seems to be phosphorylated earlier than the monomeric form following the addition of EGF and [gamma-32P]ATP. On the basis of these results, it is concluded that receptor oligomerization is an intrinsic property of the occupied EGF receptor and that it may play a role in the activation of the kinase function and the subsequent transmembrane signaling process.     

Evolution of dinosaur epidermal structures

Spectacularly preserved non-avian dinosaurs with integumentary filaments/feathers have revolutionized dinosaur studies and fostered the suggestion that the dinosaur common ancestor possessed complex integumentary structures homologous to feathers. This hypothesis has major implications for interpreting dinosaur biology, but has not been tested rigorously. Using a comprehensive database of dinosaur skin traces, we apply maximum-likelihood methods to reconstruct the phylogenetic distribution of epidermal structures and interpret their evolutionary history. Most of these analyses find no compelling evidence for the appearance of protofeathers in the dinosaur common ancestor and scales are usually recovered as the plesiomorphic state, but results are sensitive to the outgroup condition in pterosaurs. Rare occurrences of ornithischian filamentous integument might represent independent acquisitions of novel epidermal structures that are not homologous with theropod feathers.     

Epidermal focussing and the light microenvironment within leaves of Medicago sativa

The light microenvironment within leaves of Medicago sativa L. cv. Armor was related to the anatomy of the epidermis. Leaf epidermal cells had a convex shape and appeared to act as lenses that focussed light within the upper region of the palisade. In leaves irradiated with collimated light, epidermal focussing was demonstrated by ray tracing, photomicrography and fiber optic probe measurements, where lens signatures were observed. No relationship was observed between the location of focal spots within the palisade and chloroplast positioning. Epidermal focussing could be largely eliminated by irradiating leaves with diffuse light or by coating their surface with a thin layer of mineral oil that closely matched the refractive index of the cell walls. Measurement of variable chlorophyll fluorescence after elimination of epidermal focussing on the adaxial leaf surface caused a 19 and 11% decrease in the initial fluorescence level (F₀) and maximum fluorescence (F_max), respectively, whereas similar measurements from the abaxial surface were twice as large. These results suggest that some of the chloroplasts within the leaf may be adapted to local high-light conditions created by the epidermis.     

Cuticular ultrastructure of Chordodes nobilii Camerano, 1901, with a comparison of cuticular ultrastructure in horsehair worms (Nematomorpha)

The cuticle of Chordodes nobilii Camerano, 1901 is composed of a proximal layer with about 30 sheets of large fibers in alternating orientations and a distal layer, which mainly forms the surface structures, the areoles. The three different types of areoles, simple, tubercle and crowned areoles, are formed mainly by material of strong and medium electron density. The tubercle areoles have a basal constriction and paired spherical structures of unknown function below the base. Irregularly distributed are paired, cushion-like structures, from which projections traverse the cuticle and run into the epidermis. In the crowned areoles, these cushions also send projections into the apical filaments. A comparison with the few ultrastructurally described cuticles from other species reveals some similarities between the cuticles of C. nobilii and Paragordius varius, making it probable that the cushion-like structures are homologous. However, Pseudochordodes bedriagae, which is more closely related to Chordodes than Paragordius, lacks the cushions. Problematic is the interpretation of different cuticular structures in Gordius. If correctly determined, areoles are present in some Gordius species and resemble in their structure areoles from other species. If areoles have to be regarded as homologous, the absence of a distal layer and areoles would have to be interpreted as a secondary reduction in species such as Gordius aquaticus.     

Epidermal ultrastructure of Anoplodium stichopi (Plathelminthes,Dalyellioida), a flatworm endosymbiotic in Stichopus tremulus (Holothurioida)

Summary The epidermal ultrastructure of Anoplodium stichopi Bock 1925 (Platyhelminthes, Dalyellioida, Umagillidae) was studied using transmission and scanning electron microscopy. The species lives in the perivisceral coelom of the aspidochirote holothurian Stichopus tremulus Gunnerus 1767. Two types of cells were observed in the epidermis of A. stichopi: ciliated cuboidal epithelial cells and nonciliated pear-shaped cells. The surface of the ciliated epidermal cells is folded into anastomosing ridges. Numerous coated vesicles are subjacent to the surface folds and mitochondria are abundant just below them. Observations indicate that A. stichopi takes up nutrients pinocytically from the coelomic fluid of the host. The ciliation of A. stichopi is sparse.