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.     

Ultrastructural studies of the mandibular glands of the minima, media and soldier ants of Atta sexdens rubropilosa (Forel 1908) (Hymenoptera: Formicidae)

The mandibular glands of Hymenoptera are structures associated with the mandibles and constitute part of the salivary glands system. Histological studies in workers of Atta sexdens rubropilosa revealed that this gland contains two portions: a secretory and a storage portion or reservoir. Both portions are connected by means of canaliculi. The object of the present work was the study of the ultratructure of the mandibular glands of minima, media and soldier ant of A. s. rubropilosa by TEM techniques. The glands, in the three castes studied, possess a reservoir, constituted by a simple pavementous epithelium surrounded by the cuticular intima and the secretory portion is constituted by cells of rounded shape. The secretory cells, mainly of minima and soldier, were rich in smooth endoplasmic reticulum. The media worker and soldier presented a large number of mitochondria, of varying shape. Well-developed Golgi complexes were also present in the soldiers. The secretory cells in minima, media and soldier were provided with collecting intracellular canaliculi, which were linked to the reservoir through the extracellular portion. The cytoplasm of the canaliculi-forming cell was poor in organelles. In the individuals of the three castes of A. s. rubropilosa, the presence of lipid secretion granules suggested, beyond the other functions, also a possible pheromonal action. The different roles executed by the different insect castes are directly dependent on the glandular products and, consequently, on the secretory cellular characteristics.     

Salivary system in leaf-cutting ants (Atta sexdens rubropilosa Forel, 1908) castes: a confocal study

The salivary system in ants is not only limited to digestory functions, but also has important role in the communication. The glands which compose the salivary complex are: the post-pharyngeal, hypopharyngeal, mandibular, and thoracic salivary gland, showing peculiar features which may vary according to the castes the individuals belong to and according to the functions they develop. The present study compared the morphological differences among the glands of Atta sexdens rubropilosa workers, males and queens focused on the organization of microfilaments and microtubules in these ants salivary system. Although the post-pharyngeal gland appeared to be more developed in queens, there were no significant gland differences among the analyzed castes. In what regards to the secretory units of the hypopharyngeal and mandibular glands, the association of F-actin with the collector duct seemed to be strong, being surrounded by a microtubules arrangement. The use of a laser scanning confocal microscopy with immunofluorescence whole mounting preparations revealed itself an efficient instrument for the understanding of the internal morphology of insects.     

Structure of the sexually dimorphic gland of Cycloramphus fuliginosus (Amphibia, Anura, Cycloramphidae)

Cycloramphus fuliginosus males (Amphibia, Leptodactylidae) have discoid glandular elevations on the abdominal inferior body region which are considered a characteristic of genus. In this work, this sexually dimorphic gland of C. fuliginosus was investigated by low vacuum scanning electron microscopy, light microscopy, and transmission electron microscopy. In this cycloramphid, the covered integument exhibits the basic structure: the epidermis, a keratinized squamous stratified epithelium with flask cells, and the dermis. The dermis is subdivided into spongious and compact dermal layers. In the iliac gland region, an aggregate of small mucous glands and larger syncytial tubuloalveolar glands occur in the spongious dermis, and they do not go beyond the Eberth-Katschenko layer limit. The adenocytes of the mucous gland produce neutral glycoproteins, in contrast to the larger tubuloalveolar glands that elaborate a proteinaceous secretion. Myoepithelial cells surround the alveoli, and play an important role in the secretion extrusion. The final secretion, elaborated by the iliac gland, is a mixture of mucus and protein. Both secretions are important to the cycloramphid biology, and may act as pheromone and/or as chemical parental care.     

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.     

Temporal and morphological differences in post-embryonic differentiation of the mushroom bodies in the brain of workers, queens, and drones of Apis mellifera (Hymenoptera, Apidae)

The mushroom bodies are structures present in the insect brain described as centers for the neural basis of learning, memory, and other higher functions. Honeybees (Apis mellifera) are insects with a sophisticated system of spatial orientation and possess well-developed learning and memory capabilities, which are associated with neural and brain structures. Thus, the present study aimed to compare the mushroom bodies during post-embryonic development and in newly emerged males, workers, and queens using light and transmission electron microscopy to examine how differential morphological characteristics are established during development. Measurements of structures were also taken in several post-embryonic developmental phases in order to evaluate size differences during the process and in the adult organs. The results show that workers, queens, and males exhibit temporal and size differences during the post-embryonic development of mushroom bodies, probably as adaptations to differences in behavior complexity. The mushroom bodies of workers are precociously formed and are larger than those of queens and drones. Thus, workers have the largest mushroom bodies resulting from differential development during metamorphosis.     

Alterations induced by the juvenile hormone in glandular cells of the Apis mellifera venom gland: applications on newly emerged workers (Hymenoptera, Apidae)

Histological and histochemical analyses were carried out in order to evaluate the influence of the topical application of a synthetic juvenile hormone on the secretory cycle and degeneration of the venom gland of Apis mellifera. Newly emerged workers received the topical application of synthetic hormone and the results were compared to the normal development of the secretory cycle in virgin and mated queens. The first worker group received the juvenile hormone diluted in hexane (2 microg/microL), the second received only 1 microL of hexane, and the third did not receive any kind of application. After the application the workers were returned to the colony and collected at the ages of 14 and 25 days of adult life. The groups with virgin queens and the other with mated queens, did not receive the treatment. The results show that the individuals treated with juvenile hormone and with pure hexane presented differences in the histological and cytochemical aspects of the secretory cells of the venom gland. The data indicate that both the juvenile hormone and hexane accelerate the activity of the secretory cycle and the degeneration of the venom gland; however, the juvenile hormone proved to be more effective than hexane.     

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.     

Venom gland of Pachycondyla striata worker ants (Hymenoptera: Ponerinae). Ultrastructural characterization

Morphological data concerning the venom gland of worker ants of Pachycondyla striata revealed that this gland consists of three distinct regions: an external secretory portion, composed by a secretory filament that bifurcates in order to give rise to other two filaments; an internal secretory portion, represented by the convoluted gland; and a storage portion, represented by a sac-shaped reservoir. The ultrastructural analysis showed that the reservoir is enveloped by a simple pavementous epithelium, coated internally with a cuticle. The external secretory portion is composed by cells forming a simple cubic epithelium, in which the apical portion presents numerous microvilli while the basal portion of the cells shows infoldings of the plasma membrane containing numerous mitochondria. The convoluted gland possesses cells of irregular morphology with nuclei containing condensed chromatin, suggesting inactivity. However, these cells are in fact undergoing secretory activity, which is probably added to the final secretion produced by the gland. The cytoplasm of these cells contains several elements distributed therein, such as ribosomes and polyribosomes, lipid droplets, and protein inclusions in the form of crystals, thus suggestive of protein storage, which would be used by the insect when metabolically required.     

Ultrastructure of the seminal vesicle and sperm storage in Panorpidae (Insecta: Mecoptera)

The histology and ultrastructure of the seminal vesicle, male accessory gland and the epididymis in Panorpa and Sinopanorpa were observed using light microscopy and scanning and transmission electron microscopy. The seminal vesicle consists of a mono-layered elongated columnar epithelium, which contains abundant electron-dense granules and secretory vesicles, and a small central lumen. In the apical region of the epithelium of the seminal vesicle, the intense secretory activity seems to be effected by means of merocrine mechanisms. The epithelium of the accessory gland is rich in rough endoplasmic reticulum and Golgi complex, and secretes seminal fluid into the lumen via both apocrine and merocrine mechanisms. The seminal vesicle is similar to the accessory gland in the epithelium structure and their secretory activity, mainly serving a secretory function rather than storing sperms. Instead, the sperms are stored in the epididymis, whose epithelium secretes nutrients into the large lumen by merocrine mechanisms. The secretory activity and function of the seminal vesicle are briefly discussed.     

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.     

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.     

Hypokinetic stress and neuronal porosome complex in the rat brain: the electron microscopic study

Porosomes are the universal secretory machinery in cells, where membrane-bound secretory vesicles transiently dock and fuse to release intravesicular contents to the outside of the cell during cell secretion. Studies using atomic force microscopy, electron microscopy, electron density and 3D contour mapping, provided rich nanoscale information on the structure and assembly of proteins within the neuronal porosome complex in normal brain. However it remains uncertain whether pathological conditions that alter process of neurotransmission, provoke alterations in the porosome structure also. To determine if porosomes are altered in disease states, the current study was undertaken for first time using high resolution electron microscope. One of pathologies that produce subtle alteration at the presynaptic terminals has been demonstrated to be hypokinetic stress. The central nucleus of amygdale is the brain region, where such alterations are mostly expressed. We have examined the width and depth of the neuronal porosome complex and their alterations provoked by chronic hypokinetic stress in above mentioned limbic region. Specifically, we have demonstrated that despite alterations in the presynaptic terminals and synaptic transmission provoked by this pathological condition in this region, the final step/structure in neurosecretion--the porosome--remains unaffected: the morphometric analysis of the depth and diameter of this cup-shaped structure at the presynaptic membrane point out to the heterogeneity of porosome dimensions, but with unchanged fluctuation in norm and pathology.     

Ultrastructural and cytochemical characterization of T1 and T2 secretory bodies from the tegument of Echinostoma paraensei

Trematodes are lined by a syncytial layer that is named the tegument and contains small mitochondria and two different kinds of secretory inclusions. The structure and size of these bodies differs among genera and species. In a previous study, we observed many secretory bodies in the tegument of Echinostoma paraensei and named these bodies the T1 and T2 secretory bodies. No previous studies analyzed the secretory bodies of trematodes from the genus Echinostoma. Thus, the aim of this work was to use electron microscopy and cytochemistry to characterize these secretory bodies and to provide a detailed ultrastructural and morphological picture of these bodies, which are found in the tegument of E. paraensei. After ultrastructural cytochemistry analysis, we showed that both the T1 and T2 secretory bodies of E. paraensei were formed by glycoconjugates. 3D reconstruction confirmed the ovoid form of T1 secretory bodies and the biconcave and thin form of T2 secretory bodies.     

Differences in mushroom bodies morphogenesis in workers,queens and drones of Apis mellifera: Neuroblasts proliferation and death

Apis mellifera is an interesting model to neurobiological studies. It has a relatively small brain that commands the complex learning and memory tasks demanded by the social organization. An A. mellifera colony is made up of a queen, thousands of workers and a varying number of drones. The latter are males, whereas the former are the two female castes. These three phenotypes differ in morphology, physiology and behavior, correlated with their respective functions in the society. Such differences include the morphology and architecture of their brains. To understand the processes generating such polymorphic brains we characterized the cell division and cell death dynamics which underlie the morphogenesis of the mushroom bodies, through several methods suitable for evidence the time and place of occurrence. Cell death was detected in mushroom bodies of last larval instar and mainly in black-eyed pupae. Cell division was observed in mushroom bodies, primarily at the start of metamorphosis, exhibiting temporal differences among workers, queens and males.     

Ultrastructure of spined conidia and hyphae of the rice false smut fungus Ustilaginoidea virens

Spined conidia and hyphae of Ustilaginoidea virens were examined by light and electron microscopy. Bright-field light microscopy showed that conidia were round to elliptical and warty on the surface with diameters approximately ranging from 3 to 5 microm. Scanning electron microscopy revealed the globose to irregularly rounded and ornamented conidia with prominent spines. The spines were pointed at the apex or irregularly curved, and approximately 200-500 nm long. Ultrastructure of spined conidia and hyphae revealed by transmission electron microscopy showed lipid globules and vacuoles in the cytoplasm enclosed by an electron-transparent cell wall. Conspicuous electron-dense spines were evident on the surface of conidia, and had obclavate or irregularly protruding shapes with varying heights along the conidial cell wall. Microfibrillar structures with stretching or branching patterns were evident in the spine matrix. Some conidia were interconnected by spines from the neighboring conidia by their extended outgrowth. Hyphae had concentric bodies that showed an electron-transparent core surrounded by an electron-dense layer. One or more intrahyphal hyphae were found in hyphal cytoplasm. The fungus is thought to form concentric bodies and intrahyphal hyphae as survival mechanisms against the water- and nutrient-deficient environments that may occur in the necrotic regions of host plants.     

Morphology and ultrastructure of the venom apparatus in the endoparasitic wasp Pteromalus puparum (Hymenoptera: Pteromalidae)

The venom apparatus of the endoparasitic wasp Pteromalus puparum (Hymenoptera: Pteromalidae) was studied with light and electron microscope and was subjected to the electrophoretic and immunohistochemical analyses. Typically its venom apparatus consists of an unbranched venom gland and a venom reservoir, which is associated with a Dufour gland. The venom gland is lined by a series of secretory units. Each secretory unit comprises a secretory cell and a duct cell. The secretory cell is associated with an end apparatus to collect its secretions into the gland lumen. Secretory cells in the venom gland are characterized by extensive rough endoplasmic reticulum and numerous electron-dense vesicles in the distal and middle parts. They also exhibit several secretory granules and vacuoles. The venom reservoir presents three distinct regions: an external layer, composed by numerous fine muscle fibers; an internal layer, represented by epithelial cell with large nucleus; and an intima portion, represented by thin and uniform organization. The morphological aspect of numerous well-developed organelles responsible for protein generation observed is in agreement with the electrophoretic and immunohistochemical results which reveal that the rich proteinaceous components are present in the venom gland and venom reservoir. The venom proteins are first mainly produced in the secretory unit of venom gland, then drained to the lumen through the end apparatus, and are finally collected and stored in the venom reservoir.     

A scanning and transmission electron microscopic study of the development of the surface structure of neuroepithelial bodies in the mouse lung

Neuroepithelial bodies (NEBs) are groups of neuroepithelial (NE) cells that are localized on mounds on the bronchiolar epithelium of the lung. The present study examined NEBs in mice ranging in age from 2 days before birth to 80 days after birth. The position and surface architecture of NEBs was examined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In foetal mice, 2 days before birth, NEBs were distinguished from the rest of the bronchiolar epithelium by a slight elevation of non-ciliated Clara-like cells arranged in a cobbestone-like pattern. The exposed surface of the NEB was identified by small protrusions with regular microvilli intermittenly located at the base of deep clefts between the Clara-like cells. The surface of the Clara-like cells had fewer and smaller microvilli and could be easily distinguished from the apical surface of the NEB. Before birth, the surface of all of the apical cells was covered by regularly placed microville, however after birth some of the more prominently positional apical cells revealed a bare patch at the centre of the portion of apical cell exposed to the lumen of the lung. As the mice aged there was an increase in the number of apical cell protusions observed with centrally positioned bare patches. These two morphologically distinct surfaces of apical cells may have separate specialized functions. The exposed surfaces of apical cells were often observed in pairs and this feature has been observed in various sensory organs providing support for chemoreceptive function. However small bright spheres resembling vesicles were frequently observed on the lumenal surface of apical cells of the centrally placed bare patch. Transmission electron microscopy confirmed the presence of vesicles on the surface of apical cells and due to their location these vesicles were thought to contain a substance secreted into the lumen of the lung by apical cells. The significance of the bare region on the apical cells is not clear in terms of the proposed chemoreceptive function usually attributed to NEBs. It may be possible that the morphological changes observed in apical cells after birth are more appropriate for secretion of than for chemoreception. This is supported by the observation in the present study of vesicles lying on the lumenal surface of the bare region of the apical cell, however the mechanism for secretion of whoel vesicles is not clear and requires further investigation.     

A morphological view of the relationship between indirect flight muscle maturation and the flying needs of two species of advanced eusocial bees

This paper describes the flight muscles changes in relation to the age/function of the adult members of the colonies of two advanced species of eusocial bees: Apis mellifera (Apini) and Scaptotrigona postica (Meliponini). Here, are reported the results obtained through transmission electron microscopy studies, first describing a general overview of the flight muscle ultrastructure and second reporting on the ultrastructural changes that occur along the life stages/functions of workers, queens and males. The workers emerge with immature flight muscles, and the maturation takes about 20 days. In contrast, queens and males emerged with more advanced muscle differentiation, similar to workers after the 20 days of maturation. In both forager workers and laying queens, flight muscles showed signs of senescence, but not in sexually mature males. The differences among life phases, individual classes and species are discussed in relation to their functions in the colony.     

Mitochondrial and peroxisomal population in post-pharyngeal glands of leaf-cutting ants after lipid supplementation

The post-pharyngeal gland (PPG) occurs in ants and some Sphecidae wasps. Among its several roles is the storage of lipids from food. In order to investigate the effect of lipids on the cell, especially on mitochondria and peroxisomes, the present study was aimed at examining the peroxisomal and mitochondrial population in the PPG of Atta sexdens rubropilosa after lipid supplementation by confocal laser scanning microscopy and transmission electron microscopy. Soybean oil provided as lipid supplement was not toxic for A. sexdens rubropilosa workers for the first 48 h and 120 h. However, the ultrastructural cytochemical analysis revealed an accumulation of lipid droplets in the PPGs of ants after lipid supplementation at 48 h and 120 h, and smaller lipid droplets in the basal membrane of the PPG epithelium, showing lipid mobilization from the PPG to the hemolymph. The lipid supplementation reduces the life expectancy of medium workers, probably due to the high lipid metabolism. Most importantly, the PPGs of medium workers of leaf-cutting A. sexdens rubropilosa is probably a specialized gland in the lipid metabolism, due to the increased mitochondrial and peroxisomal population inside cells after lipid supplementation; participation of peroxisomal population in the β-oxidation of long chain fatty acids into smaller chains and participation of mitochondrial population in the β-oxidation of fatty acids for energy, or mobilization of lipid derivatives from the PPG to hemolymph, a process that requires energy. However, the hypothesis that the PPGs convert lipids from food in aldehydes and/or hydrocarbons must be better investigated.