The Harderian gland of the Cheesman's gerbil (Gerbillus cheesmani ) of the Kuwaiti desert

The Harderian gland is a large orbital structure. Several functions have been ascribed to the gland such as lubrication of the eye, a source of pheromones, thermoregulartory lipids and photoprotective secretions and a part of the retinal-pineal axis. In the present study, the Harderian gland of the Cheesman's gerbil, Gerbillus cheesmani, is described for the first time. The gland is located around the posterior portion of the eyeball. The gland is compound tubular, surrounded by a thin connective tissue capsule. Only one secretory epithelial cell type was recognized, characterized by the presence of lipid vacuoles and cytoplasmic slashes in high numbers; the former being more concentrated towards the apical part while the latter being more concentrated towards the central and basal parts. Some of the cytoplasmic slashes contained electron dense filamentous structures. Similar structures were observed in the lipid vacuoles. Thus, a functional relationship between the cytoplasmic slashes and the lipid vacuoles is suggested. A unique structure was observed, termed dome-like cells, located between the epithelial cells and the basement membrane. These cells were characterized by the extensive presence of pleomorphic mitochondria and compact lamellae of granular endoplasmic reticulum (GER) in the form of finger prints. The gland was found to be actively secreting porphyrins as well as lipids. Cellular debris was also seen in the tubular lumina. Myoepithelial cells with their spindle shape and elongated nuclei were evident between the basement membrane and the secretory epithelium. Sparse interstitial tissue was observed in-between the gland tubules of both male and female gerbils. Macrophages, dendritic melanocytes and lymphocytes are the most represented cellular components of the interstitium. Further studies are required to investigate the function of the dome-like cells as well as the role of lymphocytes in the rodents Harderian gland.     

Histology,histochemistry, and ultrastructure of the harderian gland of the snake Coluber viridiflavus

Analyses of the histology, histochemistry, and ultrastructre of the Harderian gland of Coluber viridiflavus prove the gland to be compound acinar and to produce a seromucous secretion. Acinar cells (type I) contain secretory granules that are composite, consisting ultrastructurally of three distinct parts that are sharply separated. They are similar to the “special secretory granules” described in the cells of the Harderian gland of the lizard Podarcis s. sicula. Some acini of the most anterior and posterior parts of the gland are mucous. Acinar cells (type II) of this type contain secretory granules that are Alcian blue/PAS positve. At the ultrastructural level, they appear homogeneous and of low density, characteristic of mucous secretions. These mucus-secreting anterior and posterior parts of the Harderian gland may by considered as regions of intial differentiation of the anterior and posterior lacrimal galnds.     

Ultrastructure of the APUD-like cells of the frog thymus gland

Within the thymus gland of the European common frog, Rana temporaria, cells with endocrine- like appearance have been found. At the ultrastructural level the most characteristic feature of their cytoplasm is the presence of secretory granules. Some cells possess irregular electron lucent granules with an eccentrically located dense core while others possess smaller electron dense granules. The cytoplasm contains also cisterns of rough endoplasmic reticulum, free ribosomes, and small mitochondria. The cells possess irregular nuclei with the pronounced nucleoli. These endocrine-like cells are connected by desmosomes with neighbouring non-granulated epithelial cells. Ultrastructural features of the cells described here resemble those seen in polypeptide hormone-secreting cells belonging to the family of cells of the APUD (Amine Precursor Uptake and Decarboxylation) series.     

Autophagy facilitates secretion and protects against degeneration of the Harderian gland

The epithelial derived Harderian gland consists of 2 types of secretory cells. The more numerous type A cells are responsible for the secretion of lipid droplets, while type B cells produce dark granules of multilamellar bodies. The process of autophagy is constitutively active in the Harderian gland, as confirmed by our analysis of LC3 processing in GFP-LC3 transgenic mice. This process is compromised by epithelial deletion of Atg7. Morphologically, the Atg7 mutant glands are hypotrophic and degenerated, with highly vacuolated cells and pyknotic nuclei. The mutant glands accumulate lipid droplets coated with PLIN2 (perilipin 2) and contain deposits of cholesterol, ubiquitinated proteins, SQSTM1/p62 (sequestosome 1) positive aggregates and other metabolic products such as porphyrin. Immunofluorescence stainings show that distinct cells strongly aggregate both proteins and lipids. Electron microscopy of the Harderian glands reveals that its organized structure is compromised, and the presence of large intracellular lipid droplets and heterologous aggregates. We attribute the occurrence of large vacuoles to a malfunction in the formation of multilamellar bodies found in the less abundant type B Harderian gland cells. This defect causes the formation of large tertiary lysosomes of heterologous content and is accompanied by the generation of tight lamellar stacks of endoplasmic reticulum in a pseudo-crystalline form. To test the hypothesis that lipid and protein accumulation is the cause for the degeneration in autophagy-deficient Harderian glands, epithelial cells were treated with a combination of the proteasome inhibitor and free fatty acids, to induce aggregation of misfolded proteins and lipid accumulation, respectively. The results show that lipid accumulation indeed enhanced the toxicity of misfolded proteins and that this was even more pronounced in autophagy-deficient cells. Thus, we conclude autophagy controls protein and lipid catabolism and anabolism to facilitate bulk production of secretory vesicles of the Harderian gland.     

Autophagy facilitates secretion and protects against degeneration of the Harderian gland

The epithelial derived Harderian gland consists of 2 types of secretory cells. The more numerous type A cells are responsible for the secretion of lipid droplets, while type B cells produce dark granules of multilamellar bodies. The process of autophagy is constitutively active in the Harderian gland, as confirmed by our analysis of LC3 processing in GFP-LC3 transgenic mice. This process is compromised by epithelial deletion of Atg7. Morphologically, the Atg7 mutant glands are hypotrophic and degenerated, with highly vacuolated cells and pyknotic nuclei. The mutant glands accumulate lipid droplets coated with PLIN2 (perilipin 2) and contain deposits of cholesterol, ubiquitinated proteins, SQSTM1/p62 (sequestosome 1) positive aggregates and other metabolic products such as porphyrin. Immunofluorescence stainings show that distinct cells strongly aggregate both proteins and lipids. Electron microscopy of the Harderian glands reveals that its organized structure is compromised, and the presence of large intracellular lipid droplets and heterologous aggregates. We attribute the occurrence of large vacuoles to a malfunction in the formation of multilamellar bodies found in the less abundant type B Harderian gland cells. This defect causes the formation of large tertiary lysosomes of heterologous content and is accompanied by the generation of tight lamellar stacks of endoplasmic reticulum in a pseudo-crystalline form. To test the hypothesis that lipid and protein accumulation is the cause for the degeneration in autophagy-deficient Harderian glands, epithelial cells were treated with a combination of the proteasome inhibitor and free fatty acids, to induce aggregation of misfolded proteins and lipid accumulation, respectively. The results show that lipid accumulation indeed enhanced the toxicity of misfolded proteins and that this was even more pronounced in autophagy-deficient cells. Thus, we conclude autophagy controls protein and lipid catabolism and anabolism to facilitate bulk production of secretory vesicles of the Harderian gland.     

Immunofluorescence-microscopic demonstration of myosin and actin in salivary glands and exocrine pancreas of the rat

Summary Actin and myosin were localized in various salivary glands (parotid, submandibular, sublingual, lingual and Harderian gland) and the exocrine pancreas of rats by indirect immunofluorescence microscopy using specific rabbit antibodies against chicken gizzard myosin and actin. A bright immunofluorescent staining with both antibodies was observed at three main sites: (1) In myoepithelial cells of all salivary glands, (2) in secretory gland cells underneath the cell membrane bordering the acinar lumen (except Harderian and mucous lingual gland), and (3) in epithelial cells of the various secretory ducts (of all glands) in similar distribution as in acinar cells. The present immunohistochemical findings in acinar cells could lend further support to a concept suggesting that myosin and actin are involved in the process of transport and exocytosis of secretory granules.Supported by grants form Deutsche Forschungsgemeinschaft (Dr. 91/1, Ste. 105/19 and U. 34/4). We thank Mrs. Ursula König, Mrs. Christine Mahlmeister and Miss Renate Steffens for excellent technical assistance.     

Ultrastructural studies of Austramphilina elongata (Cestoda,Amphilinidea)

Summary The fine structure of larval Austramphilina elongata is described using serial semithin and ultrathin sections. Densely packed germ cells with many ribosomes and mitochondria and with large Golgi complexes fill the middle third of the body. Some necrotic nuclei were observed near the anterior end. The neodermis consists of a subepidermal syncytium connected to pericarya in the parenchyma by means of cytoplasmic processes containing peripheral microtubules; electron-dense ovoid bodies condense in these processes. Myoblasts are connected to muscle fibres by means of cytoplasmic connections rich in mitochondria. Twelve (exceptionally eleven) type I gland cells containing large secretory granules and extensive granular endoplasmic reticulum are located in the dorso-posterior part of the body; they open through 12 (or 11) discrete ducts into an anterior invagination of the tegument which is covered by epidermis and not connected to the outside. Ten type II gland cells containing elongate secretory granules with regularly arranged longitudinal microtubules are located ventral to the type I cell bodies; they open on a ventral papilla a short distance behind the anterior end. Ten type III gland cells containing irregularly round-oval secretory granules with coiled microtubules are located anterior and ventral to the type I gland cells; they open through five discrete ventro-anterior openings on each side of the body. Ducts of all gland cells have mitochondria and microtubules. The spermatozoon has a basic pattern of two axonemes, each with a single central filament, a mitochondrion (mitochondria), and a row of surface microtubules interrupted by the axonemes. In the tips of epidermal cilia, doublet 1 and doublets adjacent to it lose their microtubules B first and close in on the central pair of filaments in a spiral fashion, enclosing an electron-dense rod. Presence of a neodermis and ultrastructure of the spermatozoon support the validity of the taxa Neodermata Ehlers and Trepaxonemata Ehlers and are strong evidence against a phylogenetic relationship of the cestodarians — cestodes with the Acoelomorpha; this is also indicated by the ultrastructure of sense receptors and epidermal ciliary rootlets.     

The mucigenous glandular mucosa in the comples stomach of two new-world camelids, the llama and guanaco

In contrast to the so-called true ruminants, the compartmentalized stomach of these camelids contained an extensive mucigenous glandular mucosa. This mucosal epithelium was studied with the light and electron microscope. Surface, foveolar, isthmic, and end-piece regions were identified. Undifferentiated cells with many free ribosomes and few mucigen granules were found in the gland isthmus. More fully differentiated mucigenous cells with fewer free ribosomes, an extensive Golgi complex and a large heterogeneous population of secretory granules were observed in the subjacent end-piece. These cells were compared with cardiac and other gastric glandular epithelia. The cells in the foveola contained a more extensive granular reticulum, a prominent Golgi complex, and large numbers of mucigen granules and mitochondria. In the upper foveolar cells, large supranuclear and narrow apical accumulations of mucigen granules were separated by an intervening mitochondrial mass. In the tall surface cells there was a diminution in the number of mucigen granules and a concomitant supranuclear massing of mitochondria. Basally, these cells were often separated by prominent intercellular spaces. Effete surface cells were also noted. These lacked desmosomal attachments and sometimes appeared partially extruded. These findings suggested that cells derived from the undifferentiated isthmic cells moved up the foveola and onto the luminal surface. During this migration, these cells appeared to undergo sequential cytologic differentiation. The possible functional significance of these differentiations was considered.     

Ultrastructure of the epithelial cells of the ventral prostate from the hopping mouse Notomys alexis

The large ventral prostate of the hopping mouse has abundant secretory units whose epithelial cells vary in height and which often have nuclei in the apical region of the cell. TEM observations indicated two epithelial cell types in which some unusual features occurred. Type A cells had granular endoplasmetic reticulum (GER) whose membranes often formed intracytoplasmic confronting cisternae. Type B cells had more fragmented and vesiculated GER with sparse ribosomes and less frequently also intracytoplasmic confronting confronting cisternae. In the latter cells, two types of granules were found, one of which was derived from the Golgi and the other possibly directly from the GER. Type A cells only had one type of granule present. A highly convoluted membrane was also found at the basal region in many of the cells. The significance of these unusual ultrastructural features has yet to be ascertained.     

Morphology of the Harderian gland of some Australian geckos

This study of the morphology, histology, histochemistry, and ultrastructure of the Harderian gland in Geckos (Squamata, Gekkota) revealed previously unreported variation. The gecko Harderian gland is unlike that of other squamates in that each cell of the secretory epithelium has both lipid and protein secretory granules. Lipid secretion has not been reported previously for the squamate Harderian gland. The structure of the protein granules resembles that described for a scincomorph lizard (Podarcis, Lacertidae). Differences between representatives of the subfamilies Gekkoninae and Diplodactylinae suggest possible phylogenetic constraints in the structure or function of Harderian glands within gekkotan lineages. The structural relationship between the Harderian gland and the lacrimal duct supports previous suggestions of a possible functional link between the Harderian gland and the vomeronasal organ. J Morphol 231:253–259, 1997. © 1997 Wiley-Liss, Inc.     

On the harderian gland of the domestic duck (Anas platyrhynchus)

Summary The parenchyma of the Harderian gland of the domestic duck consists of numerous tubular terminal portions, lined by a simple columnar epithelium. Its secretory surface is increased by intratubular folds. Within the cytoplasm of the epithelial cells secretory granules are observed. Polysaccharides of different nature are demonstrated. Strikingly, all centrally located cells contain a periodate reactive mucin. The successive administration of the PAS reaction and of Alcian Blue reveals the coexistence of acid and neutral mucins in the same cells. A metachromatic reaction of the mucosubstances at pH 1.0 was observed and the presence of acid sulfated groups in the Harderian gland, as demonstrated byAlcian Blue at pH 0.5, thereby confirmed. There was no glycogen reaction.The author wishes to thank Prof. Dr. W. Kühnel for his assistance and introduction to the topic for his dissertation. His thanks also go to Prof. Dr. G. Petry, and Prof. Dr. E. Roosen-Runge of the University of Washington, Seattle, USA, for their interest and suggestions.     

Ultrastructural observations on the organogenesis of the posterior silk gland of the silkworm, Bombyx mori

In the early stages of development (0 to 48 hr after organogenesis) the posterior silk gland cells of the silkworm, Bombyx mori, have characteristics of undifferentiated cells, that is, there are a number of free ribosomes in the cytoplasm and development of rough endoplasmic reticulum (rER) and Golgi bodies is very poor. Mitotic cells are frequently found. At ～ 60 hr when differentiation of the silk gland to the posterior, middle, and anterior divisions is completed, mitotic cells were no longer observable and the posterior silk gland is now composed of two rows of cells regularly packed forming a tubular structure. Differentiation of the cytoplasm is, however, not yet apparent and only a slight proliferation of rER can be observed. At 84 to 144 hr, proliferation of rER and transformation of rER from lamellar to vesico-tubular configuration are observed and Golgi vacuoles begin to enlarge. Just before hatching, the ultrastructures of cells are very similar to those of the later stage of the fifth instar when fibroin is synthesized extensively; the cytoplasm is filled with vesico-tubular rER, Golgi bodies, free secretory granules of fibroin, and mitochondria. Fibroin is probably synthesized, transported, and secreted in a manner similar to that in the fifth instar larvae.     

Ultrastructure of the parathyroid gland of the japanese lizard in the spring and summer season

The ultrastructure of the parathyroid glands of adult Japanese lizards (Takydromus tachydromoides) in the spring and summer season was examined. The parenchyma of the gland consists of chief cells arranged in cords or solid masses. Many chief cells contain numerous free ribosomes and mitochondria, well-developed Golgi complexes, a few lysosome-like bodies, some multivesicular bodies and relatively numerous lipid droplets. The endoplasmic reticulum is mainly smooth-surfaced. Cisternae of the rough endoplasmic reticulum are distributed randomly in the cytoplasm. Small coated vesicles of 700-800 Å in diameter are found occasionally in the cytoplasm, especially in the Golgi region. The chief cells contain occasional secretory granules of 150-300 nm in diameter that are distributed randomly in the cytoplasm and lie close to the plasma membrane. Electron dense material similar to the contents of the secretory granules is observed in the enlarged intercellular space. These findings suggest that the secretory granules may be discharged into the intercellular space by an eruptocrine type of secretion. Coated vesicles (invaginations) connected to the plasma membrane and smooth vesicles arranged in a row near the plasma membrane are observed. It is suggested that such coated vesicles may take up extracellular proteins. The accumulation of microfilaments is sometimes recognized. Morphological evidence of synthetic and secretory activities in the chief cells suggests active parathyroid function in the Japanese lizard during the spring and summer season.     

Fine Structure of the Esophagus of Pratylenchus penetrans

The fine structure of the esophagus of Pratylenchus penetrans is described. The gland lobe is syncytial and contains two types of nuclei: three large nuclei with little chromatin, and more numerous smaller nuclei with large amounts of chromatin. Some of the smaller nuclei are associated only with glandular tissue, whereas others are part of nerve ceils within the esophagus. Clusters of free ribosomes, rough endoplasmic reticulum, and numerous mitochondria occur in the lobe region where the secretory granules are formed. No Golgi bodies were observed. On the basis of these observations, possible differences in the mechanism of secretory granule formation between plant-parasitic nematodes are discussed. Several other minor differences between the fine structure of other plant-parasitic nematodes previously examined and that of P. penetrans are also noted.     

Fine structure of the corpuscles of stannius in the toadfish.

The micro-anatomy of the corpuscles of Stannius of the toadfish, Opsanus tau, an aglomerular marine teleost, has been studied by light and electron microscopy. The corpuscles are composed of extensively anastomosed cords of epithelial cells which maintain intimate contact with blood capillaries. Most of the epithelial cells contain acidophilic granules which also show a positive reaction with the periodic acid-Schiff technique and aldehyde fuchsin. On the basis of fine structural criteria, three cell types can be recognized. The granular cells contain abundant quantities of granular endoplasmic reticulum, ribosomes, Golgi apparatus with prosecretory granules, coated vesicles, polymorphic mitochondria with lamellar cristae, filaments, microtubules, a cilium, a variety of lysosome-like dense bodies, glycogen particles, lipid droplets, secretory granules and intranuclear lipid-like inclusions. One variety of agranular cell (type I) is characterized by the total absence of secretory granules, but it contains large amounts of granular endoplasmic reticulum and ribosomes, conspicuous profiles of Golgi apparatus, coated vesicles and sometimes an abundance of glycogen. Another variety of agranular cell (type II) has poorly developed cytoplasmic organelles. The perivascular space between the capillary and parenchyma contains connective tissue cells and abundant nerve fibers. The different types of epithelial cells observed in the corpuscles of Stannius of this fish may represent functional stages of the secretory cycle in a single cell type.     

Effects of hypergravity environment on the parathyroid gland of the propranolol-treated golden hamster

The fine structure of the parathyroid glands of propranolol-treated hamsters subjected to 5 x gravity environment was studied. In the parathyroid glands of the propranolol-treated hamsters exposed to hypergravity environment, the volume density occupied by the Golgi complexes and cisternae of the granular endoplasmic reticulum was increased as compared to that of propranolol-treated hamsters and was decreased as compared to that of hamsters exposed to a hypergravity environment but was almost similar to that of control hamsters. In addition, many chief cells contained rich free ribosomes, abundant mitochondria and some secretory granules located in the peripheral cytoplasm. These findings suggest that the parathyroid gland which may be suppressed by treatment of propranolol and stimulated in response to a hypergravity environment indicates the secretory activity of the control parathyroid gland.     

小地老虎雄性附腺细微结构和功能及高温的影响

本文通过光镜、电镜和生化分析等方法,研究了小地老虎Agrotis ypsilon(Rottemberg)雄性附腺的细微结构和功能,结果表明：(1)雄性附腺是一对管状腺,基段粉红色,中段桔红色,端段乳白色。形态分化在蛹前期完成,分泌功能在羽化后4天进入旺盛期;(2)附腺细胞属蛋白质合成型,具有旺盛的合成蛋白质的能力,胞内含有致密的粗面内质网和游离核糖体颗粒,大量的分泌液泡均匀地分布在细胞质中;(3)顶浆分泌和局部分泌是腺体的二种主要分泌方式, 前者分泌的颗粒物为糖蛋白性质(Pas阳性),后者吩泌的网状物为非糖蛋白性质(Pas阴性),二者在腺腔内呈有规则的放射状排列“4”雄性附腺分泌物具有种的特异性,小地老虎、棉铃虫和粘虫等夜蛾科昆虫分泌物的蛋白电泳谱带存在明显的种间差异,高温(32℃)抑制了雄性附腺分泌某些蛋白质的能力,从而改变精液的成分。     

Effects of cadmium on the ultrastructure of the mouse parathyroid gland

Electron-microscopic studies were made on the parathyroid gland of cadmium-treated mice. Most chief cells in treated mice were rich in free ribosomes and secretory granules compared to the control mice. In the parathyroid gland after 3 and 4 weeks of administration of cadmium, interdigitations between adjacent chief cells became more complex than in the control mice. In most chief cells of the parathyroid gland after 2, 3 and 4 weeks of administration of cadmium, the Golgi complexes associated with numerous prosecretory granules were better developed than in the control mice. These ultrastructural appearances suggest that parathyroid gland cellular activity was stimulated in response to cadmium treatment.     

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.     

The effect of aging on the rat submandibular gland: an ultrastructural, cytochemical and biochemical study

The effect of aging on the rat submandibular gland was studied by using ultrastructural, ultrastructural cytochemical and biochemical techniques. There was an age-related clumping of the nucleolar-associated and peripheral chromatin in many of the acinar cells and a decrease in the number of cisternae of rough endoplasmic reticulum. Many aged acinar cells were binucleated. There was also an age-related increase in pigment granules throughout the gland. These membrane bound granules consisted of a lipid droplet and an associated dense cap which had a granular matrix and pigment droplets. The lead capture method for acid phosphatase activity demonstrated that activity was associated with the granular matrix of the dense cap. These results were correlated with the age-related increase in acid phosphatase activity as determined by colorimetric procedures. There was an age-related increase in the number of cells characterized by small secretory granules. These cells were found as part of the intercalated ducts or at the junction of the duct with the acini. Oncocytes were also found as part of the parenchyma of the aged submandibular gland. These cells were characterized by the pleomorphic mitochondria that fill their cytoplasm. Occasionally, cells that possessed extraordinary numbers of mitochondria and small secretory granules were also observed. The determinations of total DNA and RNA revealed and age-related decrease in RNA while there was no significant change in the concentration of DNA.