Developmental anatomy of the tailed frog (As cap hits truei): a primitive frog with large eggs and slow development

The embryonic and early larval development of Ascaphus was studied by culturing embryos at 11 °C. Twenty stages of normal development are morphologically defined by using a standard staging system for anuran amphibians.
Cleavage stages are distinctive. The first cell division is unique because two separate furrows develop and then fuse to form a single cleavage plane. The large, yolky cytoplasm continues to influence cell divisions, and a blastula develops with very large vegetal cells and small, irregular shaped animal cells. There is a conspicuous, translucent blastocoel in the late blastula, and during gastrulation this structure is displayed forward by the internal migration of cells. Except for slight differences in proportions, the gastrula and neurula stages resemble the typical anuran pattern. Circulation (stage 20) begins when large vitelline veins develop on the yolk sac and direct the return of blood flow to the heart.
At hatching, the tadpole has a large yolk sac, little skin pigmentation, and no gills. Mouthparts and opercular coverings of the branchial arches develop slowly. Hind limb buds develop at the base of the tail, but they are soon concealed as a skin membrane develops and covers these structures.
The reproductive strategy for this primitive and highly aquatic frog is unusual: there is a small number of large eggs that develop slowly and produce tadpoles that continue this slow developmental process.     

Histology of melanic flank and opercular color pattern elements in the firemouth cichlid,Thorichthys meeki

Dark melanic color pattern elements, such as bars, stripes, and spots, are common in the skin of fishes, and result from the differential distribution and activity of melanin‐containing chromatophores (melanophores). We determined the histological basis of two melanic color pattern elements in the integument of the Firemouth Cichlid, Thorichthys meeki. Vertical bars on the flanks were formed by three layers of dermal melanophores, whereas opercular spots were formed by four layers (two lateral and two medial) in the integument surrounding the opercular bones. Pretreatment of opercular tissue with potassium and sodium salts effectively concentrated or dispersed intracellular melanosomes. Regional differences in epidermal structure, scale distribution, and connective tissues were also identified. J. Morphol. 2013. © 2013 Wiley Periodicals, Inc.     

Changes in the ventral dermis and development of iridophores in the anadromous sea lamprey, Petromyzon marinus, during metamorphosis: an ultrastructural study.

The ultrastructural changes that take place in the ventral dermis along with the development of iridophores were examined in the anadromous sea lamprey, Petromyzon marinus, during metamorphosis. There is a disruption of all components of the ventral dermis and a reformation that results in a structure very similar to that prior to metamorphosis. Although not a dermal component, a layer of iridophores develops directly beneath the dermis during late metamorphosis. The dermal endothelium is lost by mid metamorphosis (stage 4) and the highly organized collagenous lamellae making up the bulk of the dermis become disrupted by the migration of fibroblasts into the region. Many of these fibroblasts are involved in the degradation of the lamellae. By stage 5 of metamorphosis some fibroblasts become highly active collagen synthesizing cuboidal shaped cells that align to form a layer above the reformed dermal endothelium. New lamellae are formed by these cuboidal cells which then divide and migrate into the lamellae where they assume the characteristic attenuated appearance of fibroblasts in the adult dermal lamellae region. Iridophores first appear during stage 5 directly beneath the dermal endothelium. Reflecting platelets develop from double membraned vesicles associated with the Golgi apparatus. By late metamorphosis, stacks of trapezoidal shaped platelets fill the cytoplasm of the iridophores. The significance of the changes in the dermis during metamorphosis are discussed. This work is part of a continuing series of studies on the connective tissues in the anadromous sea lamprey.     

STRUCTURE AND FORMATION OF THE COLLAR IN CHOANOCYTES OF TETILLA SERICA (LEBWOHL), DEMOSPONGE

There are two types of collar in the choanocytes of adult Tetilla serica : one type is a continuous cytoplasmic tube and the other consists of discontinuous microvilli. The former is found in the small flagellated chamber and is considered to belong to a young choanocyte in the process of differertiation. To confirm this idea, very young choanocytes which are about to differentiate the collar were examined during embryogenesis.
The youngest choanocytes are noticed forming aggregations of small cells in 3-day larvae. Around the flagellum in each choanocyte, there is a depression which will become wider. At first, the collar is observed as a ring of cytoplasm; next this extends outward and becomes thinner, and finally it divides into microvilli. The microvillous collar is formed by the opening of vesicles and fusion of their membranes. These vesicles are considered to be derived from the Golgi complex. The process of collar formation through fusion of vesicles is discussed.     

Shift of Chloride Cell Distribution during Early Life Stages in Seawater-Adapted Killifish, Fundulus heteroclitus

The shift of chloride cell distribution was investigated during early life stages of seawater-adapted killifish (Fundulus heteroclitus). Chloride cells were detected by immunocytochemistry with an an-tiserum specific for Na(+), K(+)-ATPase in whole-mount preparations and paraffin sections. Chloride cells first appeared in the yolk-sac membrane in the early embryonic stage, followed by their appearance in the body skin in the late embryonic stage. Immunoreactive chloride cells in the yolk-sac membrane and body skin often formed multicellular complexes, as evidenced by the presence of more than one nucleus. The principal site for chloride cell distribution shifted from the yolk-sac membrane and body skin during embryonic stages to the gill and opercular membrane in larval and later developmental stages. Our observations suggest that killifish embryos and newly-hatched larvae could maintain their ion balance through chloride cells present in the yolk-sac membrane and body skin until branchial and opercular chloride cells become functional.     

An histochemical and ultrastructural analysis of the dermal chromatophores of the variant ranid blue frog

Integument from blue and green areas of the variant blue frog were analyzed biochemically for pteridines and carotenoids. Solvent extraction and absorption spectrophotometry indicated that β carotene was greatly reduced in the blue skin, and present in high quantities in the green skin of the blue frog. Thin layer and paper chromatography indicated that the pteridines were almost totally lacking in the blue skin, and present in normal quantities in the green skin of the blue frog. Light and electron microscopy indicated that the xanthophore pigment cells were either greatly altered or absent from the blue integument and present in the green integument. The fine structure of the xanthphores of the green integument contained the normal ultrastructural components of xanthopores found in regular green integument. The blue integument contained an abnormal cell type that occupied the position in the dermal chromatophore unit normally held by the xanthophores. The possibility of these cells being abnormal xanthophores or some other cell type is discussed.     

Ontogenesis and ultrastructure of seminal vesicles of the catfish,Clarias gariepinus

The ontogenesis and structural characteristics of the seminal vesicles in Clarias gariepinus (sharptooth catfish) were studied by light and electron microscopy and are described in detail. The seminal vesicles, beginning as simple protrusions from the vas efferentia, becomes more complex with age. Their distal ends become fingerlike and the bases form palm-like extensions. Juvenile male organs do not reveal any signs of seminal vesicles although spermatogenic tissue is already well delineated. The developing gonads contain clusters of large cells, close to the sperm duct and cysts of the testis, from which seminal vesicles are formed. Secretory epithelium lines the tubules of the seminal vesicles and becomes columnar as the tissue matures. Electron micro-graphs of these epithelial cells reveal two types of cells: opaque cells and cells with very vacuolized cytoplasm. Dense pinocytotic vesicles are present between the membranes of neighbouring seminal tubules and apical cell membranes facing the lumen. Maturation and onset of secretion by the secretory cells is accompanied by morphological changes. Protruding cylindrical cells become shortened, modified to cuboidal, rounded cells that send tubular extensions into the lumen. In the final stage of differentiation, only connective tissue membranes supporting the tubule walls remain intact. At the points of contact between the testis, seminal vesicles, and sperm duct, the epithelia of these organs often become confluent. The distal parts of the seminal vesicles, rarely contain sperm; during spawning sperm accumulated in the proximal tubules of the vesicles. © 1994 Wiley-Liss, Inc.     

Anatomy and ultrastructure of dermal glands in an adult water mite,Teutonia cometes (Koch, 1837) (Acariformes: Hydrachnidia: Teutoniidae)

Organization of dermal glands in adult water mites Teutonia cometes (Koch, 1837) was studied using light-optical, SEM and TEM methods for the first time. These glands are large and occur in a total number of ten pairs at the dorsal, ventral and lateral sides of the body. The slit-like external openings of the glands (glandularia) are provided with a cone-shaped sclerite, and are combined with a single small trichoid seta (hair sensillum), which is always situated slightly apart from the anterior aspect of the gland opening. Each gland is formed by an epithelium encompassing a very large lumen (central cavity) normally filled with secretion that stains in varying intensity on toluidine blue stained sections. The epithelium is composed of irregularly shaped secretory cells with an electron-dense cytoplasm and infolded basal portions. The cells possess a large irregularly shaped nucleus and are filled with tightly packed slightly dilated cisterns and vesicles of rough endoplasmic reticulum (RER) with electron lucent contents. Dense vesicles are also present in the apical cell zone. Some cells undergo dissolution, occupy an upper position within the epithelium and have a lighter cytoplasm with disorganized RER. Muscle fibers are regularly present in the deep folds of the basal cell portions and may serve to squeeze the gland and eject the secretion into the external milieu. The structure of these dermal glands is compared with the previously described idiosomal glands of the same species and a tentative correlation with the glandularia system of water mites is given. Possible functions of the dermal glands of T. cometes are discussed.     

Ultrastructural and immunocytochemical detection of keratins and extracellular matrix proteins in lizard skin cultured in vitro

The present study shows the localization of epidermal and dermal proteins produced in lizard skin cultivated in vitro. Cells from the skin have been cultured for up to one month to detect the expression of keratins, actin, vimentin and extracellular matrix proteins (fibronectin, chondroitin sulphate proteoglycan, elastin and collagen I). Keratinocytes and dermal cells weakly immunoreact for Pan-Cytokeratin but not with the K17-antibody at the beginning of the cell culture when numerous keratin bundles are present in keratinocyte cytoplasm. The dense keratin network disappears after 7-12 days in culture, and K17 becomes detectable in both keratinocytes and mesenchymal cells isolated from the dermis. While most epidermal cells are lost after 2 weeks of in vitro cultivation dermal cells proliferate and form a pellicle of variable thickness made of 3-8 cell layers. The fibroblasts of this dermal equivalent produces an extracellular matrix containing chondroitin sulphate proteoglycan, collagen I, elastic fibers and fibronectin, explaining the attachment of the pellicle to the substratum. The study indicates that after improving keratinocyte survival a skin equivalent for lizard epidermis would be feasible as a useful tool to analyze the influence of the dermis on the process of epidermal differentiation and the control of the shedding cycle in squamates.     

Neural ectoderm,neural crest,and placodes: Contribution of the otic placode to the ectodermal lining of the embryonic opercular cavity in Atlantic cod (Teleostei)

Development of neural ectoderm, neural crest, and otic placode with special reference to a new placodal derivative, the ectodermal lining of the opercular cavity, is described in a teleost fish, the Atlantic cod Gadus morhua, from a stage-by-stage examination of embryonic development. The ectodermal lining of the opercular cavity forms by invagination of the otic placode. The neural plate “infolds” by a wave of cellular rearrangement that transforms the neural plate into a neural rod. This transformation creates a distinct dorsal ectodermal cell layer. When the neural rod is arranged as monostratified columnar cells in the forebrain and midbrain, dorsal ectoderm at the midbrain level thickens lateral to the neural rod to form a cell cluster—the presumptive neural crest and placode. Upon migration of the neural crest from the postoptic midbrain, the dorsolateral area of the dorsal ectoderm thickens and segregates from the neural crest as a placode that is continuous with the presumptive lens placode. As the neural crest migrates from the hindbrain, this placode extends along the hindbrain as a single continuous cluster of cells. At the onset of formation of the lens placode, this continuous placode becomes the placode in the postoptic area of the midbrain and separates into the otic placode at the hindbrain. The otic placode gives rise to the otic neuromast and probably the otic lateral line nerves rostrally and to the ectodermal cell lining of the opercular cavity and otic vesicles caudally. The opercular cavity forms by invagination of the otic placode, creating an internal lumen lined by ectoderm that becomes continuous with evaginated endodermal pharyngeal cells. Free neuromasts are observed along the trailing edge of the external opening of the opercular cavity, which lies horizontally, ventral to the otic vesicles. As embryos develop to hatching, the opening rotates and takes up a vertical position. The adult opercular apparatus, including associated bones and muscles, forms during larval stages. The otic neuromast may be a remnant of neuromasts in the spiracle organ. The spiracle opening lies between the mandibular and hyoid arches, whereas the opercular cavity opens between the hyoid and the first branchial arches. The spiracle opening is, therefore, not homologous with the external opening of the opercular cavity, although the cell lining of the spiracle opening may be of placodal origin. J Morphol 231:231–252, 1997. © 1997 Wiley-Liss, Inc.     

Centrosome-dependent anisotropic random walk of cytoplasmic vesicles

We approach the problem of an apparently random movement of small cytoplasmic vesicles and its relationship to centrosome functioning. Motion of small vesicles in the cytoplasm of BSC-1 cells was quantified using computer-assisted microscopy. The vesicles move across the cytoplasm frequently changing their directions with negligible net displacement. The autocorrelation function for consecutive velocities of individual vesicles becomes indistinguishable from zero in 10s. Variance in the displacement is proportional to time. The motion of vesicles is anisotropic: It has diffusivity along the radii drawn from the centrosome several times higher than the tangential diffusivity. This anisotropy is abolished by ultraviolet microbeam irradiation of the centrosome when the microtubule array loses radial structure. We conclude that the motion of the vesicles in the cytoplasm can be described as diffusion-like random walk with centrosome-dependent anisotropy. The present analysis quantitatively corroborates the 'trial and error' model of vesicular transport.     

Ultrastructure of the nymphal integument in the camel tick Hyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae)

The ultrastructure of the nymphal integument in the ixodid tick Hyalomma (Hyalomma) dromedarii is compared for stages of development during and after feeding, and up to the first step of molting, apolysis. The integument comprises a cuticular layer and underlying epidermal cells. The body cuticle, which consists of both sclerotized and non-sclerotized parts, is divided into an outer, thin epicuticle, and an inner, thick, fibrillar procuticle. Pore canals in the procuticle are continuous with wax canals which traverse the epicuticle. As feeding progresses, the parallel, extensible epicuticular folds disappear due to the gut filling with ingested blood. The procuticular zone, however, becomes subdivided into an exocuticle, similar to the previously seen procuticle, and a lamellate endocuticle. Pore canals lose their parallel pattern and appear to have become deformed by stretching of the cuticle. The flat epidermal cells grow asynchronously during feeding; their cytoplasm becomes packed with well-developed rough endoplasmic reticulum (RER), while the cell apices project long microvilli extending deep into the procuticle. The RER undergoes ultrastructural changes indicating synthetic activity. Dense material released through the microvilli may serve to lyse the endocuticle and thus cause separation of the cuticle from the epidermis during apolysis. The lysed area, the exuvial cavity, is filled with lysed components which are probably withdrawn by endocytosis into the apical coated vesicles which appear in epidermal cells. Two types of integumental glands, which may participate in wax production, are observed in this study. The ultrastructure of their previously undescribed cuticular ducts is described, in addition to other hypodermal structures including epidermis-muscle attachments and sensory receptors.     

The tessellated pattern of dermal armour in the Heterostraci

The earliest and most primitive heterostraeans possessed a tessellated carapace. Isolated dentine tubercles scattered in the skin formed primordia around which concentric rings of further tubercles were laid down. This pattern of growth produced a characteristic terrazzo. From this stage the gradual elimination of tesserae can be traced in several groups. Beginning with areally growing or cyclomorial tesserae, the individual units appear simultaneously or synchronomorially, thereafter they become fused into a system of large discrete plates. Finally these synehroriomorial units appear earlier and earlier in ontogeny with progressively wider zones of cyclomorial growth being added on to them. Thus a pattern of cyclomorial plates is eventually produced.
In the psammosteids there was a redevelopment of tesserae so that the latest stages were comparable to the very early stages, although the tesserae were developed on an already existing pattern of large plates.
The possible origin of the tessellated pat tern of dermal armour is discussed. The apparent macular nature of dermal structures is not considered to be an inherent property of skin but instead due to simple physico-chemical factors.     

Light and scanning microscopic studies of integument differentiation in the grass snake Natrix natrix L. (Lepidosauria,Serpentes) during embryogenesis

We analysed the differentiation of body cover in the grass snake (Natrix natrix L.) over the full length of the embryo's body at each developmental stage. Based on investigations using both light and scanning electron microscopes, we divided the embryonic development of the grass snake integument into four phases. The shape of the epidermal cells changes first on the caudal and ventral parts of the embryo, then gradually towards the rostral and dorsal areas. In stage V on the ventral side of the embryo the gastrosteges are formed from single primordia, but on the dorsal side the epidermis forms the scale primordia in stage VII. This indicates that scalation begins on the ventral body surface, and spreads dorsally. The appearance of melanocytes between the cells of the stratum germinativum in stage VII coincides with changes in embryo colouration. The first dermal melanocytes were detected in stage XI so in this stage the definitive skin pattern is formed. In the same stage the epidermis forms the first embryonic shedding complex and the periderm layer begins to detach in small, individual flakes. This process coincides with rapid growth of the embryos.     

Brassica napus pollen development during generative cell and sperm cell formation

Summary Brassica napus pollen development during the formation of the generative cell and sperm cells is analysed with light and electron microscopy. The generative cell is formed as a small lenticular cell attached to the intine, as a result of the unequal first mitosis. After detaching itself from the intine, the generative cell becomes spherical, and its wall morphology changes. Simultaneously, the vegetative nucleus enlarges, becomes euchromatic and forms a large nucleolus. In addition, the cytoplasm of the vegetative cell develops a complex ultrastructure that is characterized by an extensive RER organized in stacks, numerous dictyosomes and Golgi vesicles and a large quantity of lipid bodies. Microbodies, which are present at the mature stage, are not yet formed. The generative cell undergoes an equal division which results in two spindle-shaped sperm cells. This cell division occurs through the concerted action of cell constriction and cell plate formation. The two sperm cells remain enveloped within one continuous vegetative plasma membrane. One sperm cell becomes anchored onto the vegetative nucleus by a long extension enclosed within a deep invagination of the vegetative nucleus. Plastid inheritance appears to be strictly maternal since the sperm cells do not contain plastids; plastids are excluded from the generative cell even in the first mitosis.     

'Nectosome': a novel cytoplasmic vesicle containing nectin in the egg of the sea urchin, Temnopleurus hardwickii

Localization of an extracellular matrix protein, Th-nectin, in the eggs and embryos of the sea urchin Temnopleurus hardwickii was examined by both immunofluorescence and immunoelectron microscopy. The protein is associated with a tubular structure packaged in rod-shaped vesicles that were designated as 'nectosomes'. In unfertilized eggs, nectosomes are distributed uniformly throughout the cytoplasm, but after fertilization, they gradually translocate to the cortical zone where they are arranged perpendicular to the plasma membrane. The migration of the nectosomes was strongly inhibited by cytochalasin B, which suggested that microfilaments play an important role in this process. Immunocytochemical and immunoblotting analyses both ascertained that nectin is secreted into the hyaline layer. Some nectosomes remain in the apical cytoplasm of dermal cells until the gastrula stage. Ultrastructural examination revealed that the accumulation of nectosomes in the oocyte cytoplasm begins quite early in oogenesis, concomitant with the accumulation of cortical vesicles.     

Integument initiation and testa development in some Cruciferae

This study has shown for the first time that the middle layer (or layers) of the outer integument is (are) of subdermal derivation in at least some taxa of the Cruciferae. The outer integument is initiated in the Cruciferae in three different ways, viz. subdermally (Brassica, Sinapis) , partly subdermally and partly dermally (Lunaria) , or completely dermally (Capsella). These differences in initiation are reflected in the structure of the mature testa. The inner integument is completely of dermal derivation and originally two cell-layers thick, but may become more than two-layered during the ovule and seed maturation by periclinal divisions of the inner cell layer. The consequences of the ontogeny of the integuments for the terminology and interpretation of the mature testa is discussed.     

The ultrastructure of the integument and proventriculus in the raptorial rotifer Cupelopagis vorax (Monogononta: Collothecaceae: Atrochidae)

Members of the sessile rotifer species Cupelopagis vorax are unusual ambush predators that live permanently attached to submerged freshwater plants. Previous light microscopical research has revealed several uncommon features in this species including a stellate‐patterned integument and an expansive foregut region called the proventriculus. In this study, we apply transmission electron microscopy to explore the ultrastructure of both the integument and foregut to determine how they differ from other rotifers. Our results reveal that the integument is covered by a thick glycocalyx and is patterned with tubercles that originate from the intracytoplasmic lamina (ICL) within the syncytial epidermis. The ICL forms an apical layer within the syncytium, is electron dense and mostly amorphous, and forms tubercles up to 2.3 μm; these tubercles probably account for the patterned appearance of the integument and are similar to what has been found in other gnesiotrochan rotifers. The basal cytoplasm is highly granular and contains two types of membrane‐bound vesicles: large ovoid vesicles (320–411 nm) with amorphous, opaque contents, and secretory bulbs (110–264 nm) with electron‐lucent cores and occasionally electron‐dense contents. Only the secretory bulbs were observed to form connections to the apical plasmalemma, and so are probably exocytotic. Internally, the proventriculus is a large distensible sack that connects the anterior pharyngeal tube to the posterior mastax. The proventricular epithelium is a thin syncytium mostly covered with a dense glycocalyx and a strong brush border of microvilli underlain by a thin terminal web. The cytoplasm contains few organelles and there is no evidence that it is either secretory or has features (e.g., ICL) that might aid in maceration. We hypothesize that the thick glycocalyx might serve a protective function against the movements of live prey and/or against enzymes released from the rotifer's gastric glands that become regurgitated during feeding.     

THE FINE STRUCTURE OF RHODOSPIRILLUM RUBRUM

The fine structure of Rhodospirillum rubrum grown under a series of defined conditions has been examined in thin sections prepared by the methods of Ryter and Kellenberger. In cells grown anaerobically at different light intensities, the abundance of 500 A membrane-bounded vesicles in the cytoplasm is inversely related to light intensity, and directly related to cellular chlorophyll content. When the chlorophyll content of the cell is low, the vesicles are exclusively peripheral in location; they extend more deeply into the cytoplasm when the chlorophyll content is high. Typical vesicles also occur, though rarely, in cells grown aerobically in the dark, which have a negligible chlorophyll content. When synthesis of the photosynthetic pigment system is induced in a population of aerobically grown cells by incubation under semianaerobic conditions in the dark, the vesicles become increasingly abundant with increasing cellular chlorophyll content, and the cells eventually acquire the cytoplasmic structure that is characteristic of cells growing anaerobically at a high light intensity. Poststaining with lead hydroxide reveals that the membranes surrounding the 500 A vesicles are indistinguishable in structure from the cytoplasmic membrane, and continuous with it in some areas of the sections. The bearing of these observations on current notions concerning the organization of the bacterial photosynthetic apparatus is discussed.