Morphology of Bufo ictericus integument (Amphibia, Bufonidae)

Bufo ictericus integument was investigated by stereoscopic, low vacuum scanning electron microscopy, transmission electron microscopy and light microscopy. The studies revealed, that the dorsal integument surface is rougher than ventral. Three types of projections are visualized: larger rounded verrucae, smaller conical cornified tubercles, and conical short spines. Prominent verrucae are observed on the dorsal surface, being flatter on the ventral surface. The tubercles are visualized only on the dorsal surface. The verrucae are separated by grooves that may contribute spreading and retention of the glandular secretion upon the integument. The pattern of the epidermal grooves is also important for water distribution, protecting the animal against desiccation. The epidermis is composed of a stratified epithelium with intraepithelial blood vessels, where keratinocytes predominate, but flask cells, and Merkel cells also occur. In the spongious dermis, cutaneous glands are visualized. The compact dermis is a series of alternating layers of bundles of collagenous fibers, and between spongious and compact dermis there are basophilic areas that correspond to Eberth-Katschenko layer. The dorsal and the ventral surfaces of B. ictericus are morphologically distinct. The integument structure is related to the physiology of each surface and represents an adaptation to habitat, reflecting a lifestyle of the animal.     

Microscopical methods promote the understanding of the integument biology of Rhinella ornata

In Rhinella ornata, the integument of different body regions was investigated using low-vacuum scanning electron and light microscopy through histochemical and immunohistochemical methods, and revealed the basic structure found in other anurans. Keratinocytes formed the keratinized squamous stratified epithelium, and flask cells occurred among the epidermal superficial layer. Just below the epidermis, the dermis was subdivided into a spongious dermis and a compact dermis. Mixed and granular glands were located in the spongious dermis, and myoepithelial cells surrounded their secretory portions. The Eberth-Katschenko (EK) layer occurred as basophilic areas between the spongious and compact dermis and throughout the spongious dermis. A series of alternating layers of bundles of collagenous fibers characterized the compact dermis, being firmly attached to a thin hypodermis. Regarding the morphological features, each integument region revealed distinct structural aspects. The dorsal integument was rougher than ventral, having conical tubercles and spines upon the verrucae. In the ventral and subgular regions, the slightly elevations are visualized, tubercles were absent but some spines occurred. The pelvic integument had a non-keratinized epidermis with an irregular profile and small poorly developed elevations. In addition, the EK layer was absent, and the presence of numerous blood vessels suggests its participation in water absorption and distribution. In the parotoid gland, the epidermis was similar to the dorsal integument; except that the reticular dermis was present, and contained three exocrine glandular types. Expression of cytokeratin was evident in the basal and intermediary layer, but absent in the cornified layer. Flask cells showed strong cytokeratin labeling. Consequently, the integument had the basic morphology, but exhibits regional characteristics, which may be related to the physiology of each surface.     

Morphological characterization of the anuran integument of the Proceratophrys and Odontophrynus genera (Amphibia, Anuran, Leptodactylidae)

The morphological characteristics of the leptodactylid integument of Proceratophrys and Odontophrynus genera were investigated by means of stereoscopic, low vacuum scanning electron and light microscopy. The integument surface of Proceratophrys boiei, Proceratophrys laticeps and Proceratophrys appendiculata exhibited several projections, while the integument of Odontophrynus americanus had rounded elevations with smooth profile. Light microscopic observations showed the basic integument morphology for all anurans, i.e., an epidermis and a dermis, which is subdivided into a spongious layer and a compact layer. The epidermis is formed by basal, intermediary and cornified layers. However, in Proceratophrys genus the cornified layer had an irregular outline, while in O. americanus the external surface was smooth. In the spongious dermis, mucous and venom exocrine glands were observed, but in O. americanus an exclusive glandular type with apocrine secretory pattern was identified. The integument morphology showed peculiar characteristics that may be helpful for genus distinction. Thus, morphological methods may be considered as an efficient means to characterize and to differentiate anuran genera.     

Histochemistry of skin glands of Trachycephalus aff. venulosus Laurenti, 1768 (Anura, Hylidae)

The skin of the adult tree frog, Trachycephalus aff. venulosus, was investigated by light microscopy. Histochemistry was carried out using various stain techniques and showed the basic integument morphology of the skin with squamous epithelium in the epidermis and connective tissue in the dermis, subdivided into spongy and compact layers. The integument observed contains two types of gland; the mucous glands and granular glands. The mucous glands are small and located in the upper layer of the stratum spongiosum of the connective tissue and contain neutral and acid sulfated glycoproteins. The granular glands are large and form a syncytial secretory compartment within the acinus, which is surrounded by smooth muscle cells. Histochemical analyses demonstrated the presence of collagen proteins in both types of gland. An unusual finding was the different densities of the granules in the granular glands. The morphological characteristics of the integument in Trachycephalus aff. venulosus was also compared with others species of Anura.     

Hyaluronan in the epidermal and the dermal extracellular matrix: its role in cutaneous hydric balance and integrity of anuran integument

New researches have revealed that hyaluronan (HA) is not a passive molecule. HA has being pointed out to participate in many processes, such as cell interactions, proliferating and migrating cell events and function as hydrate agent. The present study was undertaken to localize HA in Bufo ictericus integument providing a better understating for the role of cutaneous HA. Paraffinized sections were stained with haematoxylin-eosin and with 1% Alcian blue 8GX at pH 1.0 and 2.5. Alcianophilic reaction was visualized in both spongious dermis and Eberth-Katschenko layer. The mucus cells of mucus glands were also stained with AB methods. Besides these histological techniques, the localization of HA was performed using the FITC-labeled HA probe labeled with fluorescein isothiocyanate. In the extracellular matrix of spongious dermis, the reaction for HA was evident, being less intense in hypodermis and in pericellular keratinocyte matrix of the cornified tubercles regions. Thus, since HA was localized in the pericellular epidermal matrix and in the spongious dermis of anuran integument, it plays an important role in hydric balance, and is essential for integument integrity and functionality.     

Ultrastructural aspects of the mandibular gland of Melipona bicolor Lepeletier, 1836 (Hymenoptera: Apidae, Meliponi) in the castes

The mandibular gland in Melipona bicolor workers and queens was studied by scanning and transmission electron microscopy. There is no difference in the gland anatomy between the castes, but the transmission electron microscopy showed variation of the cellular ultrastructure according to the secretory phase of the gland in both castes. Smooth endoplasmic reticulum was abundant in the secretory cells of physogastric queens, indicating that these cells produce lipid secretion that is stored in granules with multi-lamellar bodies. Mitochondrial variations during the cell secretory cycle indicates their participation in the lipid synthesis. After secretion, release in the reservoir lumen through the collecting canals, the secretory cells contain many myelinic bodies, indicative of cellular regression.     

Dermal collagen organization in Bufo ictericus and in Rana catesbeiana integument (Anuran, Amphibian) under the evaluation of laser confocal microscopy

Collagen structural organization plays an important role in the mechanical property of the vertebrate integument. Bufo ictericus and Rana catesbeiana integument was investigated by light microscopy and laser confocal microscopy. Collagenous elements of the dermis were statistical analyzed. The integument is formed by the keratinized squamous stratified epidermis supported by the dermis that is subdivided into the spongious layer with a loose arrangement, and the compact layer formed by collagenous fibers arranged compactly in a criss-crossed manner. Thick collagenous columns have a perpendicular trajectory, and are formed by the assembling of alternating collagenous lamellae in both animals. Short intercolumns of collagenous fibrils connecting collagenous lamellae obliquely or transversally are observed in R. catesbeiana dorsal integument. The present study provides evidences that B. ictericus and R. catesbeiana integument has well-organized compact dermis, constituted by collagenous lamellae in a plywood manner. The integument organization is in contrast to the literature in some aspects. This dermal arrangement is important to the biomechanical property of both anuran integuments.     

Ultrastructural study of the salivary glands of the sugarcane spittlebug Mahanarva fimbriolata (Stal, 1854) (Euhemiptera: Cercopidae)

Spittlebugs are insects that suck sap from plants and regurgitate saliva containing toxic enzymes into the leaves. As a consequence, the conductive channels are blocked resulting in dry leaves, thus giving a burned aspect to the plantation. This work performed ultrastructural analyses of the salivary glands of the sugarcane spittlebug Mahanarva fimbriolata, since these organs produce the enzymes that are injected into the plants, thus being responsible for the economic losses in the production of sugarcane. Three kinds of secretory cells are found in principal gland, forming the lobules I-IV. The main differences among these cells relate to size, morphology and electron density of the secretory vesicles. The accessory glands contain different secretory vesicles to those in the principal gland. Muscular cells are found around the entire gland. The different secretory vesicles found in both principal and accessory glands indicate that the gland produces different substances or that the secretion in the interior of cells passes through a maturation process.     

Dehiscent organs used for defensive behavior of kamikaze termites of the genus Ruptitermes (Termitidae,Apicotermitinae) are not glands

During Isoptera evolution, the caste of soldiers disappeared in some Apicotermitinae termites as in the Neotropical Ruptitermes. Paired dorsolateral structures located between the metathorax and abdomen of foraging workers of Ruptitermes were previously denominated dehiscent glands, and are responsible for releasing an adhesive secretion that immobilizes enemies, causing their death. In this study, we investigated the morphology of dehiscent organs of workers of Ruptitermes reconditus, Ruptitermes xanthochiton, and Ruptitermes pitan and also second instar larvae of R. reconditus using light, laser scanning confocal, and transmission electron microscopy. Additionally, we performed a preliminary protein analysis using SDS-PAGE to further characterize the secretion of Ruptitermes dehiscent organs. Our results showed that the dehiscent organs do not exhibit the typical characteristics of the exocrine glandular cells class I, II or III of insects, suggesting that they constitute a new type of defensive organ. Thus, the denomination dehiscent gland was not used but dehiscent organ. Dehiscent organs in larvae are formed by fat body cells. In workers, dehiscent organs are composed by compact masses of cells that accumulate a defensive secretion and are poor in organelles related to the production of secretion. Since the dehiscent organs are not glands, we hypothesize that the dehiscent organs originate from larval fat body. The defensive secretion may have been produced at younger developmental stages of worker or the defensive compounds were absorbed from food and accumulated in the worker fat body. Histochemical techniques and SDS-PAGE revealed that the secretion of Ruptitermes dehiscent organs is constituted mainly by a protein of high molecular weight (200 kDa). In conclusion, the dehiscent organs are extremely different from the exocrine glands of termites and other insects described until now. In fact, they seem to be a specialized fat body that is peculiar and exclusive of Ruptitermes termites.     

The intramandibular gland of leaf-cutting ants (Atta sexdens rubropilosa Forel 1908)

The eusociality developed in Hymenoptera and Isoptera is driven by an efficient interaction between exocrine glands and jointed appendages, both in close interaction with the environment. In this context, the mandible of ants plays an important role, since, in addition to being the main jointed appendage, it possess glandular functions. As an example we might name the two glands associated with the mandible: the mandibular and the intramandibular glands. The intramandibular gland is found inside the mandible and consists of a hypertrophied secretory epithelium and secretory cells in the mandible's lumen. The secretion of the secretory epithelium is liberated through intracuticular ducts that open at the base of hairs at the mandible's surface. The secretion of the intramandibular gland (epithelium and secretory cells) reacted positively to tests for the detection of polysaccharides and proteins, thus suggesting that it consists of glycoproteins. The ultrastructure of the secretory epithelium presents variations related to the developmental stage of the individual, showing a large number of ribosomes and microvilli close to the cuticle in young individuals, while in the older specimens it was possible to note the formation of an intracellular reservoir. These variations of secretory epithelium, as also the interaction between the cellular groups inside the mandible, are important information about this gland in leaf-cutting ants.     

The venom gland of queens of Apis mellifera (Hymenoptera, Apidae): morphology and secretory cycle

The venom gland of queens of Apis mellifera was examined through light and transmission electron microscopy and subjected to electrophoretic analyses. Virgin queens exhibited prismatic secretory cells containing large amounts of rough endoplasmic reticulum with dilated cisternae, open secretory spaces, numerous vacuoles and granules scattered in the cytoplasm, and spherical nuclei with numerous nucleoli. The secretion produced was non-refringent under polarized light and the electrophoretic analysis of glandular extracts revealed five main protein bands. In mated queens, the venom gland exhibited a high degree of degeneration. Its secretion was refringent under polarized light and one of the main bands was absent in the electrophoretic pattern obtained. The morphological aspects observed are in agreement with the function of this gland in queens, given that virgin queens use venom in battles for the dominance of the colony, a situation that occurs as soon as they emerge, while fertilized queens rarely use venom.     

Morphology of the sternal gland in workers of Coptotermes gestroi (Isoptera, Rhinotermitidae)

The sternal gland is considered the only source of trail pheromones in termites. The morphology of the sternal gland was investigated in workers of Coptotermes gestroi using transmission and scanning electron microscopy. The results showed a small bilobed gland at the anterior part of the fifth abdominal sternite. The cuticular surface of the sternal gland showed a V-shaped structure with two peg sensilla in elevated socket and various campaniform sensilla. Pores and cuticular scale-like protuberances also occur in the glandular area. The ultrastructure showed a gland composed of class 1 cells and two different types of class 3 cells distinguished by location, different size and electron-density of secretory vesicles. Small class 3 cells (type 1) of the anterior lobe are inserted among class 1 cells and have weakly electron-dense vesicles associated with mitochondria, glycogen and smooth endoplasmic reticulum. The class 3 cells (type 2) of posterior lobe showed many round electron-lucent vesicles of secretion, abundant free ribosomes and a well-developed Golgi apparatus. Each class 3 cell is connected to the cuticle by a cuticular duct constituted by the receiving canal and the conducting canal. The secretion of class 1 cells is stored in an inner subcuticular reservoir that is delimited by the microvilli of these cells. This inner reservoir is large and crossed by the campaniform sensilla and ducts of two types of class 3 cells that open outside of the insect body. An exterior reservoir also is present between the fourth and fifth sternite. The complex structure of the sternal gland suggests multicomponents for the trail pheromone in the worker of C. gestroi.     

Ultrastructure of the skin melanophores and iridophores in paddlefish, Polyodon spathula

The ultrastructure of melanophores and iridophores of Polyodon spathula has been examined by transmission electron microscopy. In the skin, two types of chromatophores, melanophores and iridophores were founded. Melanophores were localized both in epidermis and dermis. Epidermal melanophores were present on the dorsal region of the trunk, sides, outer surface of the operculum and rostrum. Iridophores were founded in the dermis from ventral skin. The cytoplasm of iridophores is filled with reflecting platelets with variable orientation. The length of the long axis of the platelets varies from 1 to 2.10 microm.     

Male reproductive system structure and accessory glands ultrastructure of two species of Triatoma (Hemiptera,Reduviidae, Triatominae)

Morphological studies in male reproductive system of insect, and accessory glands functions, can contribute to the knowledge of reproductive aspects in Triatominae. The present study aims to describe the structure of male reproductive system under scanning electron microscopy, and the ultrastructure of accessory glands during the process of maturation, in T. brasiliensis and T. melanica of different ages.These system consist of two testis, two vas deferens, two seminal vesicles, four pair of accessory glands and one ejaculatory duct. The four mesodermal male accessory glands of T. melanica and T. brasiliensis, present secretion storage in the firsts days of adult life in starved males. During the maturation process of male accessory glands of T. brasiliensis and T. melanica, granules with different sizes, shapes and electron density were seen. In T. melanica small vacuoles are released into the gland lumen in one-day-old males, however, after three and five days few granules are found in the cell cytoplasm. The secretory granules in T. brasiliensis increase in size and amount, in the five days old adult. The secretory pathway in the male accessory glands of the two species seems to be merocrine and apocrine suggesting that different substances are being produced at different times. The cell microvilli in T. brasiliensis are longer and narrower, these can be absent in some regions of the cell surface of T. melanica. Maturation of male accessory glands in the triatomine species studied occurred without adult blood meal. Its function can led to new insights for the comprehension of reproductive aspects in Triatominae.     

Characterisation of CART-containing neurons and cells in the porcine pancreas,gastro-intestinal tract,adrenal and thyroid glands

Background  

The peptide CART is widely expressed in central and peripheral neurons, as well as in endocrine cells. Known peripheral sites of expression include the gastrointestinal (GI) tract, the pancreas, and the adrenal glands. In rodent pancreas CART is expressed both in islet endocrine cells and in nerve fibers, some of which innervate the islets. Recent data show that CART is a regulator of islet hormone secretion, and that CART null mutant mice have islet dysfunction. CART also effects GI motility, mainly via central routes. In addition, CART participates in the regulation of the hypothalamus-pituitary-adrenal-axis. We investigated CART expression in porcine pancreas, GI-tract, adrenal glands, and thyroid gland using immunocytochemistry.     

Morphology and histology of the digestive system of the vector leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae)

The vector leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae) is an important economic pest that is a serious threat to wheat in Northwest China, causing great losses to agricultural production by transmitting wheat blue dwarf (WBD) in a persistent circulative manner. Studies on morphology and ultrastructure of the digestive system were made using light, scanning and transmission electron microscopy. The gross morphology of the alimentary canal consists of esophagus (foregut), midgut, hindgut, and special filter chamber. The esophagus, a narrow and slender tube, runs through the whole thorax extending into the filter chamber and connecting with the anterior midgut. The midgut consists of three major regions, anterior, middle and posterior midgut. The anterior midgut is composed of exceptionally thick epithelial cells surrounding a large lumen. Numerous well-developed microvilli occur at the basal region of the epithelium. A large number of electron-lucent and lipid-like vesicles are observed under the microvilli. The posterior midgut is smaller than the anterior midgut in diameter. Numerous different concentric circular bodies are characteristic of the posterior midgut. The hindgut arises from the posterior midgut at the boundary of the filter chamber. It extends downward toward the anus where the hindgut enlarges to form the rectum. The rectum is formed by various cells typical of epithelium, whose nuclei are relatively smaller than those of the midgut. Relatively large muscle cells are present at the well-defined and thick basement membrane. There are two pairs of Malpighian tubules. Numerous mitochondria and lamellar rough endoplasmic reticulum in the cells of the anterior segment of the Malpighian tubules; the sub-anterior segment of the tubules resemble the distal segment which are wave-like, amount vesicles in the cells; numerous tightly packed large vesicles contain two types of brochosomes in the inflated segment of the Malpighian tubules. The salivary system of P. striatus contains one pair of salivary glands, which are made up of the principal and accessory gland. The principal glands are divided into anterior, mid- and posterior lobes. About nineteen acini are observed in the principal glands of P. striatus, and vary in structure and colour. Five cellular types are found in the principal glands by transmission electron microscopy, which are rich in secretory granules, modified in size, electron-dense and shape. Rough endoplasmic reticulum in the shape of vesicular and lamellar, mitochondria and Golgi complexes are observed. The accessory gland has only an oval or a rod-shaped acinus.     

Glycosaminoglycans and glycoconjugates in the adult anuran integument (Lithobates catesbeianus)

Glycosaminoglycans (GAGs) from the integument of Lithobates catesbeianus were biochemically characterized and histochemically localized. Moreover, carbohydrate distribution was investigated using conventional and lectin histochemistry at light microscopy. Hyaluronan (HA), dermatan sulfate (DS) and a heparanoid were found in the integument. Sulfated and carboxylated GAGs were visualized in the Eberth-Katschenko (EK) layer, in the mucous glands, in the hypodermis as well as in the mast cells. Furthermore, glucose and galactose were identified in the integument through thin layer chromatography (TLC) assays. N-Acetyl-β-glucosamine residues were identified in the mucous glandular cells, between the corneum and spinosum strata, in the subepidermal region, and in the EK layer. N-Acetyl-galactosamine residues were evident in the EK layer, corresponding to a residue of the dermatan sulfate chain, which may be related to the collagenous fiber arrangement. These glycoconjugates occurred as secretory glandular products and as dermal structural elements. Moreover, HA and DS are the predominant GAGs in the L. catesbeianus integument. Considering the importance of glycoconjugates, they play a significant role to the integrity of the skin, providing mechanical support for integument cells. In addition, they are important to the water regulation mechanisms, since L. catesbeianus is preferably aquatic.     

Fine structure of Mytella falcata (Bivalvia) gill filaments

Bivalve filter feeders are sessile animals that live in constant contact with water and its pollutants. Their gill is an organ highly exposed to these conditions due to its large surface and its involvement in gas exchanges and feeding. The bivalve Mytella falcata is found in estuaries of Latin America, on the Atlantic as well as the Pacific Coast. It is commonly consumed, and sometimes is the only source of protein of low-income communities. In this study, gill filaments of M. falcata were characterized using histology, histochemistry and transmission electron microscopy for future comparative studies among animals exposed to environmental pollutants. Gill filaments may be divided into abfrontal, intermediate and frontal zones. Filaments are interconnected by ciliary discs. In the center of filaments, haemocytes circulate through a haemolymph vessel internally lined by an endothelium and supported by an acellular connective tissue rich in polysaccharides and collagen. The abfrontal zone contains cuboidal cells, while the intermediate zone consists of a simple squamous epithelium. The frontal zone is composed of five columnar cell types: one absorptive, mainly characterized by the presence of pinocytic vesicles in the apical region of the cell; one secretory, rarely observed; and three ciliated with abundant mitochondria. All cells lining the filament exhibit numerous microvilli and seem to absorb substances from the environment. PAS staining was observed in mucous cells in the frontal and abfrontal zones. Bromophenol blue allowed the distinction of haemocytes and detection of a glycoprotein secretion in the secretory cells of the frontal region. The characteristics of M. falcata gill filaments observed in this study were very similar to those of other bivalves, especially other Mytilidae, and are suitable for histopathological studies on the effect of water-soluble pollutants.     

Application of NMR microscopy to the morphological study of the silkworm,Bombyx mori,during its metamorphosis

Zero (ZQ) and double (DQ) quantum 2D chemical shift selective and spin-echo 3D NMR imaging at microscopy resolution, has been applied to the morphological study of silkworm, Bombyx mori, during its metamorphosis. Attention has been focused on the evolution of the internal structure of the insect during its postembryonal life occurring through the larval, pupal and adult development. A major objective of this work was the characterization of the silk glands, responsible for synthesis and secretion of fibroin and sericin, through the changes of distribution and mobility of water, by imaging the water protons during postembryonal life stages. Moreover, alanine deriving from silk gland proteins was imaged during the last life stage of Bombyx mori.     

Volume-weighted mean volume of the submandibular gland acini in male and female diabetic rats

Mouth dryness is one of the complications of diabetes mellitus but there has been little work on morphological changes of the salivary glands. In the present study, the effects of diabetes mellitus on the serous and mucous acini of submandibular gland of male and female rats, 4 and 12 weeks after diabetes induction were studied. Male and female rats were divided into experimental and control subgroups. Diabetes was induced to experimental rats by streptozotocin. At the end of 4 and 12 weeks, the submandibular glands were removed, random sections obtained and volume-weighted mean acini volume was estimated by point-sampled intercepts method. The data revealed that volume reduction occurred only in serous acini in both male and female rats 12 weeks after diabetes induction and the others remained unchanged. The present research using stereological methods demonstrates that diabetes make some morphological changes in serous acini, the main exocrine part of the rat submandibular gland.