Ultrastructural study of two glandular systems in the proboscidial glandular epithelium of Riseriellus occultus (Nemertea, Heteronemertea)

 Two different types of glandular system in the proboscidial epithelium of Riseriellus occultus have been investigated by transmission electron microscopy. As expected, most of the epithelial cells are glandular in nature. With regard to differences in the ultrastructure of these gland cells and in the formation and morphology of their secretory granules, we have categorized and described four types of gland cell, indicated as G₁, G₂, G₃, and G₄. Each gland cell has a completely intraepithelial body characterized by a prominent nucleus, developed rough endoplasmic reticulum, Golgi complexes, and numerous secretory granules at different stages of maturation. These four types of gland cell appear associated in pairs forming numerous glandular systems of two types (A, B). These glandular systems are restricted to the ventral surface of the proboscis and are scattered irregularly throughout its length. Each glandular system consists of two gland cells of different types. The gland cell necks in each glandular system extend together to the epithelial surface; they protrude onto this and form a papilla where they open in a common area. The epithelial supportive cells adjacent to the glandular systems have long, stout microvilli which have a core of tonofilaments. These tonofilaments gather into dense bundles which pass vertically through the supportive cells and attach to the extracellular matrix underlaying the cells by hemidesmosomes. Moreover, a single sensory process stands close to each papilla. The ultrastructural morphology of the type A glandular systems suggests that they have an adhesive function operating in a similar way to that of the duo-gland adhesive systems in other invertebrate groups, although they are not homologous with these. The spatial arrangement of the secreted products of the type B glandular systems suggests that these may contribute to increasing the grip of the proboscis on the prey. The secretory granules (=pseudocnids) of the type G₃ gland cells are very likely an autapomorphy of the Anopla, providing a character by which the relationships within the Nemertea can be evaluated. Accepted: 9 October 1997     

The C cell system and experimental thyroid apudomas in the rat: the combined action of 2-acetylaminofluorene and chlorpromazine in inducing systemic medullary type tumors. An ultrastructural study

Previous chronic administration of chlorpromazine (CPZ) before the start of the 2-acetylaminofluorene (2-AAF) action and during the cancerogenic process systematically facilitate the growth of apudome-like medullary tumors inside the thyroid gland. Ultrastructural studies of normal C cells, adenomatous C cells and cancerous C cells derivated from the normal cellular type show various specific transformations of few intracytoplasmic organelles. In the Golgi apparatus, the path of main maturation of calcitonin-like granules seems the same in the normal and in the adenomatous cells. On the contrary, the amount of calcitonin-like granules falls in the tumoral extensive tissue even though the golgian activity turns towards a high abnormal proliferation of clathrin vesicles or coated vesicles. This new fact is directly related with numerous extensions and repairs of much damage of the neoplastic cell membranes. Finally, the large medullary tumors (showing many atypic mitosis in glandular neoplastic C cell-type) are deeply penetrated by extensive and irregular vegetative nervous fibers.     

The tarsal gland of ixodid ticks <Emphasis Type="Italic">Ixodes persulcatus</Emphasis> and <Emphasis Type="Italic">Ixodes ricinus</Emphasis> (Mesostigmata,Ixodidae)

A complicated multicellular gland is situated in all the leg tarsi, occupying from one third to half the segment. The glandular cells form a single-layer sack; the inner surface of the gland cavity is covered with the multi-layer membrane. Cuticular rods (“sinews”) of muscles moving the claw pass inside the gland cavity. The glandular cells are characterized by the presence of numerous microvilli on their apical surfaces and by the presence of secretory vacuoles. The basal part of each secretory cell is characterized by accumulations of lipid vacuoles and glycogen granules. Problems concerning the possible role of tarsal gland in the production of the trace pheromone are discussed.     

AN ELECTRON MICROSCOPE STUDY OF MATURE AND DIFFERENTIATING PANETH CELLS IN THE RAT, ESPECIALLY OF THEIR ENDOPLASMIC RETICULUM AND LYSOSOMES

In an electron microsope study, the morphology of mature Paneth cells from the small intestine of adult rats is compared with that of differentiating Paneth cells from young rats 2 to 4 weeks old. All mature cells exhibit a marked polarity similar to that of other exocrine gland cells and contain a well developed endoplasmic reticulum, an elaborate Golgi complex, and numerous large secretory granules; they also possess an abundance of lysosomes. The most conspicuous occurrence in the process of differentiation is the development of the endoplasmic reticulum. The most immature Paneth cells possess an endoplasmic reticulum of the vesicular type, which, during maturation, is replaced by the characteristic lamellated ergastoplasm of the mature cell. At a certain stage of differentiation the cavities of the developing cisternae show numerous communications with the perinuclear space, suggesting an outgrowth of the ergastoplasm from the nuclear envelope. Furthermore, the cavities and the perinuclear space at this particular stage contain a material which shows a remarkable intrinsic periodicity. An identical periodicity was exhibited by material contained in Golgi cisternae and secretory granules. Lysosomes are also present in the differentiating cells.     

The amylase-secreting cells of the stomach of the scallop, Pecten maximus: ultrastructural, immunohistochemical and immunocytochemical characterizations

(1) alpha-amylase was extracted and purified from the stomach/digestive gland complex of the scallop Pecten maximus and an anti-serum was induced against the purified amylase by rabbit immunization. (2) The anti scallop amylase was used to localize the amylase-secreting cells in the stomach of Pecten maximus by immunofluorescence and immunogold labelling. The amylase-secreting cells are glandular cells particularly numerous in the main sorting area of the stomach. Their secretory granules were found strongly positive for anti-amylase. Three types of glandular cells were observed, actually corresponding to the three stages of the glandular-cell activity, synthesis, secretion and excretion. (3) The synthesizing cell shows the characteristic features of a protein-synthesizing cell: a conspicuous nucleolus and abundant granular endoplasmic reticulum. In the secretory cell, the secretory granules are formed by the Golgi apparatus and accumulate in the apical part of the cell. The secretory cell is filled with two types of secretory granules which are released in the stomach lumen by apocrine excretion. (4) The present study brings the first demonstration of the synthesis and extracellular release of amylase by glandular cells of the stomach epithelium of a bivalve.     

The harderian gland of the frog, Rana esculenta, during the annual cycle: histology, histochemistry and ultrastructure

The Harderian gland in Rana esculenta has been studied during the annual cycle at the histological, histochemical and ultrastructural levels. The Harderian gland has an acinar structure and is the only orbital gland in anuran amphibia. It develops at the medial corner of the orbit from the conjunctival epithelium at the premetamorphic stage. In the adult the glandular secretion reaches a maximum during the months of July and August, drops in September and resumes slowly from October onwards. The secretion is seromucoid and the secretory granules are released into the acinar lumen, mainly by exocytosis. Porphyrins were not detected. No sexual dimorphism was observed in the glandular cells. The resumption of secretory activity in October and the enhancement of secretion in May are marked by the appearance of "blue nuclei" (Mallory stain) in a relatively high percentage of glandular cells. This unusual blue colour, using the Mallory stain (by which nuclei stain red), disappears after digestion of paraffin sections with RNAase, but not with DNAase and trypsin. The blue staining may, therefore, indicate an increased amount of nuclear RNA. The Harderian gland in the frog most probably serves to lubricate and moisten the eye in the absence of the lacrimal gland. However, the gland may also represent an immunoactive organ owing to the presence of numerous mast cells and plasma cells in the interacinar spaces.     

An ultrastructural study of the prostate gland of megascolicid earthworm Lampito mauritii (Kinberg)

The ultrastructure of prostate gland of Lampito mauritii revealed two types of secretory cells. Type 1 cells with a broad basal region and a long apical region contain electron dense oval secretory granules with an increased density at the core region. Numerous electron lucent granules with fine filamentous and electron dense amorphous materials also occur at the basal region of these cells. Type 2 cells contain electron lucent mucous-like secretory granules. This cell type contains exceptionally large Golgi complexes having 20-23 stacked cisternae. Both cell types open into a common lumen and numerous microtubules are visible at the apical end. Junctional complexes, such as desmosomes and septate junctions, are observed in this glandular tissue.     

Secretory and basal cells of the epithelium of the tubular glands in the male Mullerian gland of the caecilian Uraeotyphlus narayani (Amphibia: Gymnophiona)

Caecilians are exceptional among the vertebrates in that males retain the Mullerian duct as a functional glandular structure. The Mullerian gland on each side is formed from a large number of tubular glands connecting to a central duct, which either connects to the urogenital duct or opens directly into the cloaca. The Mullerian gland is believed to secrete a substance to be added to the sperm during ejaculation. Thus, the Mullerian gland could function as a male accessory reproductive gland. Recently, we described the male Mullerian gland of Uraeotyphlus narayani using light and transmission electron microscopy (TEM) and histochemistry. The present TEM study reports that the secretory cells of both the tubular and basal portions of the tubular glands of the male Mullerian gland of this caecilian produce secretion granules in the same manner as do other glandular epithelial cells. The secretion granules are released in the form of structured granules into the lumen of the tubular glands, and such granules are traceable to the lumen of the central duct of the Mullerian gland. This is comparable to the situation prevailing in the epididymal epithelium of several reptiles. In the secretory cells of the basal portion of the tubular glands, mitochondria are intimately associated with fabrication of the secretion granules. The structural and functional organization of the epithelium of the basal portion of the tubular glands is complicated by the presence of basal cells. This study suggests the origin of the basal cells from peritubular tissue leukocytes. The study also indicates a role for the basal cells in acquiring secretion granules from the neighboring secretory cells and processing them into lipofuscin material in the context of regression of the Mullerian gland during the period of reproductive quiescence. In these respects the basal cells match those in the epithelial lining of the epididymis of amniotes.     

Ultrastructure and maturation of a sex pheromone gland in the female German cockroach, Blattella germanica

A volatile sex pheromone is produced in an adult female-specific gland located on the anterior of the last abdominal tergite of the female German cockroach, Blattella germanica (L.). In this area, the cuticle forms deep depressions in which a large number of cuticular orifices are located. The cuticular orifices are connected to secretory cells via cuticular ducts surrounded by duct cells. The pheromone gland exhibits a clear developmental maturation in relation to sexual maturation of the female. The secretory cells of a newly formed gland in the imaginal female are small and contain few secretory vesicles. The amount of extractable pheromone in the gland is low on day-0 but it increases with age and peaks on day-6. The secretory cells in a mature day-6 gland are characterized by a large number of electron-lucid secretory vesicles. abundant RER and SER, a large nucleus and a long, convoluted end apparatus which is lined with numerous microvilli. The contents of the secretory vesicles are exocytosed into extracellular reservoirs at the base of microvilli and then transported to the cuticular surface through the long ducts. The supportive function of the duct cell in the glandular organization and developmental regulation of the gland are discussed.     

Ultrastructural changes in the pars distalis of the maturing male rat

Anterior pituitary glands of male rats (2, 3, 5, 8, 12, 25, 36, 52, 56, and 62 days of age) were processed for electron microscopy. During early postnatal stages secretory cells are found in various stages of differentiation and comparatively few secretory granules are seen. Nuclei are mostly irregular, and the nucleo-cytoplasmic ratio is large. Many free ribosomes are present; the endoplasmic reticulum is generally sparse and the Golgi complex small or invisible. Cells are of variable shape, and numerous cytoplasmic processes project into large intercellular spaces. Many electron-dense cells which often contain myelinlike figures are seen. Lysosomes and lysosomal precursors are frequently found in secretory cells, predominantly in somatotrophs, of all immature glands. Mitotic figures are numerous in early stages after brith and decrease in number as the gland grows in size. A gradual increase in cytoplasmic volume with concomitant differentiation of cytoplasmic components as well as accumulation of secretory granules, accompanied by loss of myelin-like figures and decrease in the number of electron-dense cells, is observed as the animal reaches the prepuberal stage. Few lysosomes are seen in cells of mature glands. At 36 days of age all secretory cells seem to have differentiated, and morphological features as well as granule content show little change until puberty is reached. Gonadotrophs attain their characteristic morphology later than other cells. Cilia are observed in all developmental stages but are relatively infrequent in the mature gland. The described ultrastructural characteristics reflect the degree of maturation as well as the functional capacities of secretory cells at particular stages of development.     

Morphological study of the development of the disseminate prostate in intact and castrated male pigs aged two to six months.

The light-microscopical and ultrastructural differentiation of the disseminate prostate was studied in 12 boars and 8 barrows, ranging in age from 8 to 28 and 12 to 27 weeks, respectively. The barrows had been castrated at the age of 8 weeks. In young boars the prostate is organized in an inner zone and an outer zone. The glandular tubules of the latter are more differentiated than those in the inner zone. Sexual maturation in the prostate is characterized by almost complete reduction of the inner zone and concomitant enlargement of the outer zone. This differentiation of the inner zone occurs progressively and is manifested by a higher density of the glandular tubules, by a decrease in the number of basal cells and by an increase in the secretory tubular cells containing more microvilli, organelles and secretory vesicles. A few weeks after castration, the characteristic morphology of the undifferentiated inner zone is found throughout the entire glandular layer. The number of glandular tubules in barrows has decreased, and their cells show minimal secretory activity, nuclear indentations and a decrease in their organelles. A further castration effect is vacuolization of the perinuclear cytoplasm in numerous glandular cells.     

Histo- and cytophysiology of the lactating mammary gland of the African elephant (Loxodonta africana)

The lactating mammary gland of the African elephant (Loxodonta africana) has been studied with a panel of morphological techniques focusing on (1) the functional changes during the secretory process, (2) proliferative process [by application of proliferating cell nuclear antigen (PCNA) immunohistochemistry] and apoptotic phenomena [by use of the TUNEL technique] in the individual lobules, and (3) components of milk and milk-fat-globule membrane. In the lactating gland, the lobules are variably differentiated; within a lobule, however, the alveoli are usually similarly differentiated. The morphology of their alveoli suggests a classification of the lobules into types 1–3. Lobules of type 1 are composed of immature tubular alveoli with mitotic figures and numerous PCNA-positive nuclei; advanced type 1 alveoli contain abundant glycogen and specific secretory granules. Lobules of type 2 are further subdivided. In type 2a lobules, the epithelial cells of the alveoli form tall apical protrusions, which in part are occupied by small lipid droplets and which are pinched off in an apocrine fashion. The number of lysosomes varies considerably. Type 2b is the most common type, with striking basal membrane foldings, abundant rough endoplasmic reticulum cisterns, large Golgi apparatus, numerous mitochondria, lipid droplets, and protein vesicles with 30- to 90-nm-wide casein micelles. The lipid droplets are pinched off with minimal amounts of cytoplasm. Type 2c is composed of alveoli with a cuboidal epithelium and few signs of secretory activity. Increasing expression of peanut-agglutinin-binding sites parallels the maturation and differentiation of the glandular cells. Type 3 lobules are marked by numerous TUNEL-positive nuclei and large lipid droplets and are apparently degenerating structures. Cytokeratin (CK) 14 is usually present in the myoepithelial cells; CK 19 and CK 7 mark ductal and immature alveolar epithelia. Milk protein content varies between 2.6% and 6.3%, and casein micelles range from 35 to 90 nm in diameter. The diameter of intra-alveolar milk fat globules ranges from 5 to 25 µm and the membranes bear a filamentous surface coat composed of membrane-anchored mucins; gel-electrophoretic analysis of these mucins from different individuals demonstrates the presence of mucin MUC 1, which is expressed with considerable genetic heterogeneity.     

Epidermal glands in Squamata: morphology and histochemistry of the pre-cloacal glands in Amphisbaena alba (Amphisbaenia)

Summary Pre-cloacal glands occur in some species of amphisbaenians. Although these glands are important in systematics, their biology and chemistry are little known. The pre-cloacal glands of Amphisbaena alba are of the holocrine type. They are made up of a glandular body and a duct. The glandular body is conical to elongate and is formed of clongatc lobules separated one from another by collagen septa. Each lobule is composed, at its periphery, of germinative cells, and within of polyhedral secretory cells, of different degrees of differentiation. The germinative cells, set on a basal lamina, are basophilic and their cytoplasm is fairly electron dense. The polyhedral cells display bulky cytoplasm, filled with spherical granules, wrapped in membranes and differing in their electron densities. Towards the lumen of the gland, these granules are increasingly eosinophilic and have an affinity for orange G. The secretion is discharged into the duct leading to the pore, which is situated in the central region of the scale. This secretion shows positive histochemical results for mucopolysaccharides and proteins. The similarity between the epidermal glands of lizards and those of A. alba raises the suggestion that the glands have equivalent functions, possibly in the course of intra- or interspecific communication.     

The formation of glandular secretion in larval Austramphilina elongata (Amphilinidea)

The ultrastructure of three types of gland cells of embryos and free-swimming larvae of Austramphilina elongata is described. Type I gland cells contain large, more or less round electron-dense granules which are formed by numerous Golgi complexes. Type II gland cells contain thread-like, membrane-bound secretory granules with longitudinally arranged microtubules inside the granules; secretory droplets are produced by Golgi complexes and the microtubules apparently condense in the cytoplasm or in the droplets. Type III gland cells contain irregular-ovoid membrane-bound granules with coiled up microtubules which have an electron-dense core; the granules are formed by secretionderived from Golgi complexes and the microtubules aggregate around and migrate into the secretion; microtubules are at first hollow and the early secretory granules have a central electron-dense region.     

An insight into the skin glands,dermal scales and secretions of the caecilian amphibian Ichthyophis beddomei

The caecilian amphibians are richly endowed with cutaneous glands, which produce secretory materials that facilitate survival in the hostile subterranean environment. Although India has a fairly abundant distribution of caecilians, there are only very few studies on their skin and secretion. In this background, the skin of Ichthyophis beddomei from the Western Ghats of Kerala, India, was subjected to light and electron microscopic analyses. There are two types of dermal glands, mucous and granular. The mucous gland has a lumen, which is packed with a mucous. The mucous-producing cells are located around the lumen. In the granular gland, a lumen is absent; the bloated secretory cells, filling the gland, are densely packed with granules of different sizes which are elegantly revealed in TEM. There is a lining of myo-epithelial cells in the peripheral regions of the glands. Small flat disk-like dermal scales, dense with squamulae, are embedded in pockets in the dermis, distributed among the cutaneous glands. 1–4 scales of various sizes are present in each scale pocket. Scanning electron microscopic observation of the skin surface revealed numerous glandular openings. The skin gland secretions, exuded through the pores, contain fatty acids, alcohols, steroid, hydrocarbons, terpene, aldehyde and a few unknown compounds.     

Concanavalin a binding and cytodifferentiation in pancreatic acinar carcinoma of rat

The binding of concanavalin A to the plasmalemma of acinar carcinoma cells was characterized by electron microscopy utilizing horseradish peroxidase. Heavy labeling due to specific concanavalin A binding was detected on the plasmalemma of undifferentiated carcinoma cells lacking zymogen maturation, neoplastic cells of intermediate differentiation with only occasional zymogen granules, and highly differentiated acinar carcinoma cells containing numerous cytoplasmic zymogen granules. The plasmalemma of acinar carcinoma cells was also compared to the normal pancreatic acinar cell plasmalemma by measurement of specific ¹²⁵I-labeled concanavalin A binding. Although only about one-third of pancreatic acinar carcinoma cells demonstrate mature zymogen differentiation, the acinar carcinoma had a full complement of normal plasmalemma receptors for ¹²⁵I-labeled concanavalin A. It is concluded that, unlike normal pancreas, the presence of concanavalin A receptors on the plasmalemma of acinar carcinoma cells is not a specific membrane marker for differentiated cells containing zymogen granules.     

Ultrastructural changes in the oviduct of the laying hen during the laying cycle

The ultrastructural changes occurring in the fully functional oviduct of Isa Brown laying hens were studied during various stages of the laying cycle. Hens were killed at different positions of the egg in the oviduct. The oviduct was lined by ciliated and non-ciliated cells (also referred to as granular cells). The granular cells in the infundibulum contributed to secretion during egg formation, whereas ciliated cells showed little evidence of secretion. Ultrastructural changes were recorded in the granular and glandular cells of the distal infundibulum. In the magnum, the surface ultrastructure revealed glandular openings associated with the ciliated and granular cells. Cyclic changes were recorded in the glandular cells of the magnum. With respect to the three observed types of glands, the structure of gland type A and C cells varied at different egg positions in the oviduct, whereas type B cells represented a different type of gland cell containing amorphous secretory granules. The surface epithelium of the isthmus was also lined by mitochondrial cells. Two types of glandular cell (types 1 and 2) were recorded in the isthmus during the laying cycle. Intracisternal granules were found in type 2 cells of the isthmus. A predominance of glycogen particles occurred in the tubular shell gland. The granular cells in the shell gland contain many vacuoles. During egg formation, these vacuoles regressed following the formation of extensive rough endoplasmic reticulum; the reverse also occurred. The disintegrated material found in the vacuoles may have been derived from the disintegrating granules. The Physiology Teaching Unit, University of New England, provided financial support to K. Chousalkar for this study.     

Organogenesis of the pituitary, adrenal, and lung at birth in the wallaby, Macropus rufogriseus

The ultrastructure of the pituitary, the adrenal, and the lung was examined in the newborn wallaby, Macropus rufogriseus. Tissue from six wallaby neonates (less than 8 hr of age), two near-term fetuses (26 days after removal of suckling pouch young [RPY]), and a two-day-old pouch young was examined; and tissue levels of cortisol in the adrenal glands of five neonates and a near-term fetus (26 days) were measured by radioimmunoassay. At birth the adenohypophysis comprised the bulk of the pituitary gland. The pars distalis was well vascularized and many cells contained electron-dense, membrane-bound granules. The adrenal glands lacked specific zones but comprised two distinct populations of cells. The cytoplasm of one cell type contained electron-dense, membrane-bound granules, similar to those observed inside catecholamine-secreting cells of the adrenal medulla; the other cell type possessed large amounts of smooth endoplasmic reticulum and mitochondria with tubulo-vesicular cristae. These features are characteristic of cells which are actively synthesizing steroid hormones. The concentration of cortisol was 0.58 ng/adrenal in the wallaby at birth. The fetal lungs near term were at the glandular stage of development, and epithelial differentiation of type I and type II pneumocytes was imminent although attenuation was not evident. The canalicular neonatal lung did not contain true alveoli, but type II pneumocytes contained osmiophilic lamellar inclusions of surfactant. The fetal pituitary and adrenal are functional at birth and are thus capable of initiating parturition and of influencing lung maturation in the fetus.     

Postnatal development of the Mongolian gerbil uterine glands

The development of gerbil uterine glands has been described. Gland formation starts about day 6 postnatally (p.n.). At day 10 p.n. the glandular lumen contains already some fluid, and the glandular cells are in an activated state. The apical shape of the glandular cells depends mainly on the width of the lumen. Secretory vesicles in early developmental stages are electron-lucent. As sexual maturity is reached, secretory granules become electron-dense. This change in secretory granular structure indicates a change in the quality of uterine gland fluid.