The secretory nature of the excretory gland cells of Stephanurus dentatus. II. Presence of hydrolytic enzymes

Enzymes catalyzing the hydrolysis of casein, N-benzoyl-l-tyrosine ethyl ester, p-nitrophenyl acetate, and l-leucyl-β-naphthylamine hydrochloride were found in extracts of the excretory gland cells of Stephanurus dentatus.     

Proteinase inhibitors from the excretory gland cells of Stephanurus dentatus. Purification and properties of three secretory proteinase inhibitors

Three proteinase inhibitors designated as I, II, and III were isolated from the excretory gland cells of the swine kidney worm, Stephanurus dentatus. The inhibitors, which were trichloroacetic acid-soluble, were purified by affinity chromatography and ion exchange chromatography. The homogeneity of each inhibitor was shown by polyacrylamide gel electrophoresis and electrofocusing. The molecular weights of the inhibitors estimated by sodium dodecyl sulfate gel electrophoresis fell within a limited range of 9300 to 9700, and the isoelectric points were 6.45, 6.20, and 5.34 for Inhibitors I, II, and III, respectively. The inhibitors formed complexes with trypsin having apparent dissociation constants (Ki) of 2.9 X 10(-11), 7.6 X 10(-11), and 6.4 X 10(-11) M, respectively. Each inhibitor inhibits the esterolytic and proteolytic activities of both trypsin and chymotrypsin. A proteinase inhibitor present in the reproductive organs, intestines, body walls, and esophagi was identical with Inhibitor II found in the excretory gland cells. Culture medium collected after 24-h incubation with adult worms contained the same three inhibitors as the excretory gland cells. These data suggest that the gland cells may secrete the inhibitors internally and externally.     

Occurrence of unusual heterogeneous lipid-containing granule-storing cells in the main excretory duct epithelium of the male mouse submandibular gland

Summary The fine structure of the main excretory duct epithelium of the male mouse submandibular glands was investigated by scanning and transmission electron microscopy. Three principal cell-types were observed: type I and II, and basal cells. This epithelium was characterized by the presence of intercellular canaliculi. Type-I cells were the most numerous. They had an abundance of mitochondria, well-developed Golgi apparatus, a few electron-lucent lipid-containing granules and poorly developed basal infoldings. These cells were also characterized by many glycogen granules throughout the cytoplasm and abundant smooth endoplasmic reticulum in the apical cytoplasm. Type-II cells were the second most numerous. Their most characteristic feature was the presence of abundant heterogeneous lipid-containing granules having acid phosphatase activity at the periphery. They were concentrated in the infra- and supranuclear cytoplasm. The granules may be derived from mitochondrial transformation and seem to be a special kind of secondary autolysosome. Type-II cells also contained abundant mitochondria throughout the cytoplasm, much smooth endoplasmic reticulum in the apical cytoplasm, a well developed Golgi apparatus adjacent to the heterogeneous lipid-containing granules and no basal infoldings. Basal cells were situated adjacent to the basal lamina. They had a large nucleus and the cytoplasm was filled with glycogen granules.     

The biochemical nature of the cell periphery of the salt gland secretory cells of fresh and salt water adapted mallard ducks

Summary Both ruthenium red and dialyzed iron techniques indicated that anionic protein-carbohydrates were associated with the plasmalemma of secretory cells in mallard salt glands. Digestion of the tissue with neuraminidase prior to dialyzed iron staining provided evidence that much of the anionic nature of this surface was due to sialic acid. These results were confirmed by biochemical assays showing that the salt-gland tissue contained a relatively high sialic-acid content and that the amount of sialic acid increased with salt-water adaptation. Possible roles of these anionic sites are discussed in relation to currently accepted hypotheses of electrolyte transport.Research supported by Public Health Service Grant AM-13705     

Ultrastructure and nature of secretory proteins in the male accessory gland of Drosophila funebris

The ultrastructural differentiation of the secretory cells and the nature of secretory proteins in the male accessory gland of Drosophila funebris have been studied by electron-microscopic and immunological methods. (1) In the pupae at $\text{1}\text{1}\text{2}$ days before eclosion, secretory products can be detected in the lumen, even though most glandular cells are at the initial phase of differentiation. At the time of eclosion both main and secondary cells are fully differentiated, but the whole set of five immunologically active proteins are detectable only on the second to third day of adult life. (2) The secondary cells contain giant protein granules, the so-called filamentous bodies, which become partially fused and the filaments assume a twisted form. Randomly dispersed filaments and closely packed filament bundles are also visible in the gland lumen. Antigenic labelling of ultrathin sections and immunoreplica electrophoresis yielded no evidence for the microtubular nature of these filaments. The secretion stored in the lumen contains in addition a large quantity of flocculent proteins which have their origin in the main cells. (3) During the period of high secretory activity in the 7-day-old male flies no vacuolization and disintegration of either the main or secondary cells have been observed. We conclude that both types of cells have the merocrine secretory mechanism. (4) Ultrastructural alterations in the glandular cells confirmed our previous observation that copulation stimulates RNA and protein synthesis.     

Reaction of the main proteolytic fraction of Schistosoma mansoni cercarial enzymes with synthetic substrates and inhibitors of proteolytic enzymes

The purified proteolytic fraction of Schistosoma mansoni cercarial enzymes (PCF) was inhibited by Diisopropylphosphofluoridate (DFP). Its esterolytic activity was not affected by either of two specific active center reagents of proteolytic enzymes: (1) 1-chloro-3-tosylamido-7-ammo-2-heptanone (trypsin) and (2) l-1-tosylamido-2-phenylethyl chloromethyl ketone (chymotrypsin). Furthermore, PCF did not destroy the biological activity of bradykinin on the isolated guinea pig ileum, nor did it release bradykinin from purified dog plasma bradykininogen.     

Cultivation of epithelia from the secretory coil of the ovine apocrine gland: Evidence of secretory cell function and ductal morphogenesis In vitro

Summary The secretory coil of the ovine apocrine gland is composed predominantly of two cell types, secretory cells lining the lumen and myoepithelial cells adjacent to the basement membrane. The glands synthesize a number of hormones and growth factors, but analysis of the functions of these molecules may be hampered by the mixing of apocrine and sebaceous secretions in the pilary canal. The purpose of this study was to isolate the glands and devise simple culture procedures to facilitate investigations of secretory cell function. The most successful approach involved microdissection of the secretory coils individually from skin biopsies and culture in Dulbecco’s modified Eagle’s medium. After 1–2 wk in medium, cell outgrowths were seen from explants. These consisted predominantly of populations of epithelial cells, many containing granules. Smaller granules were usually concentrated around the cell nuclei and accumulated lipophilic dyes. Large granules were unreactive. Western analysis showed that cells in culture synthesized nerve growth factor-like peptides, a feature consistent with one of the functions of the gland in vivo. When isolated secretory coils were explanted to culture dishes coated with matrigel, highly compact, multilayered masses of cells grew out. Subsequently, tubular structures formed. The observations suggest that some differentiated functions of gland cells were retained in vitro and that the procedures described provide a system for the study of apocrine secretions in isolation from those of other skin glands.     

The relation between solid cell nests and C cells of the thyroid gland

Summary Thyroid tissue of 300 routine autopsies was processed in a standardized manner. So-called solid cell nests (SCN) were found in 21 patients (7 %). These cases were investigated carefully by serial step sectioning. In order to explore the correlation of SCN to the C-cell system, the sections were stained by silver impregnation and the immunoperoxidase method. Morphometric analyses revealed a significant increase in the density of C cells in the proximity of the SCN. With progressive distance from the SCN, the C-cell density decreased and reached normal values. In 30 % of the cases argyrophilic and calcitonin-positive cells were found lying within the SCN. Occasionally, mixed follicles could be discerned: These were lined on the one side by a multilayered squamous epithelium, on the other side by normal monolayered cubic follicular epithelium, and contained a peculiar granular material. In one case, SCN were associated with intrathyroid portions of the parathyroids and adult adipose tissue, in a second case with adipose tissue only. Most probably SCN are vestiges of the ultimobranchial body and should be interpreted as such, despite the fact that other authors have expressed different views. The lack of disturbances in the calcium metabolism of the patients and the absence of medullary carcinoma in their family histories led us to interpret locally confined C-cell hyperplasia not as reactive nor premalignant, but rather as normal.     

Interstitial and parenchymal cells in the pineal gland of the golden hamster

Summary A combined thin-section/freeze-fracture study was performed on the superficial pineal gland of the golden hamster, comparing the parenchymal and interstitial cells of this animal with those previously investigated in rats. In contrast to rats, no gap junctions and gap/tight junction combinations could be found between pineal parenchymal cells of the hamster. Furthermore, the interstitial cells of the hamster pineal gland were found to have large flat cytoplasmic processes, which abut over large areas equipped with tight junctions. In thin sections, profiles of interstitial cell processes were seen to surround groups of pinealocytes. Interstitial cells and their sheet-like, tight junction-sealed processes thus appear to delimit lobule-like compartments of the hamster pineal gland. Because the classification of the interstitial cells is uncertain, the expression of several markers characteristic of mature and immature astrocytes and astrocyte subpopulations has been investigated by indirect immunohistology. Many of the non-neuronal elements in the pineal gland are vimentin-positive glial cells, subpopulations of which express glial fibrillary acidic protein (GFA) and C1 antigen. The astroglial character of these cells is supported by the lack of expression of markers for neuronal, meningeal and endothelial cells. M1 antigen-positive cells have not been detected.Supported by a grant from Deutsche Forschungsgemeinschaft (Scha 185/9-2)     

The differentiation of the decidua and the distribution of metrial gland cells in the pregnant mouse uterus

Summary A study was made with the light microscope of the cellular changes involved in the formation of the decidua in the pregnant mouse uterus up to day 11 of pregnancy. The differentiation sequence was similar to that found by previous workers but the morphology and development of the basal zone is described in more detail. In addition, the morphology of glycogen rich cells in an area termed the lateral decidual zone is described. The distribution of granulated metrial gland cells and their precursors is described. These cells are first found in the uterine stroma before the blastocyst has implanted. Later they occur in the decidua and in the circular smooth muscle zone beneath the mesometrial triangle prior to the formation of the metrial gland. Granulated metrial gland cells are also found in the maternal blood spaces of the implantation site.     

Effects of age on morphology,protein synthesis and secretagogoue-evoked secretory responses in the rat lacrimal gland

This study investigated changes in the morphology and protein synthesis and protein and peroxidase secretion due to peptidergic and aminergic stimulation from rat lacrimal gland acinar cells of 3–5, 9, 12, 20 and 24 month old rats. There was a marked reduction in the presence of Golgi apparatus in the acinar cells of glands from the 24 month old rats coupled to dilatation and degeneration of rough endoplasmic reticulum, when compared to that in the acinar cells of glands from 3–5 and 12 month old rats. Following incorporation of tritiated leucine for 360 min (6 h), the amount of newly synthesised protein in acinar cells of the 12 month old rats was significantly (p < 0.01) higher than that in the acinar cells of 3–5 month old animals. However, at 20 months the amount of newly synthesised protein in these acinar cells was significantly (p < 0.01) reduced to less than that in acinar cells of both the 3–5 and 12 month old animals. Immunohistochemical and immunofluorescence studies identified the presence of substance P (SP), vasoactive intestinal peptide (VIP), histamine and 5-hydroxytryptamine (5-HT) in the lacrimal glands of 3–5 month old rats. Stimulation by either SP, VIP, histamine or 5-HT resulted in significant increases in total protein output and peroxidase release from acinar cells of the 3–5 month old rats. However, all responses to the secretagogues were reduced with ageing from 3–5 to 24 months of age. The results indicate that ageing is associated with alteration in the ability of acinar cells to synthesise and secrete proteins.     

Ultrastructural studies on the secretory cavities ofCitrus deliciosa ten. I. Early stages of the gland cells differentiation

Summary Secretory cavities ofCitrus deliciosa seem to originate from a pair of meristematic cells (an epidermal cell and a second one placed under it). These cells undergo successive divisions resulting in the formation of a globular/oval gland situated in the parenchyma, and a conical stalk, which joins the gland with the epidermis. The young gland consists of a central group of polyhedral cells ensheathed by layers of radially flattened cells.During the early differentiation stages of the gland cells a close association of cytoplasmic microtubules with various organelles is observed. Plastids increase progressively in number and size and their matrix locally contains tubular networks accompanied by small oil droplets. In peripheral cytoplasm numerous myelin-like lomasomes have been observed. Peripheral cells of the gland are gradually modified from the inner cells following a different developmental pattern. Their walls become thicker and plastids do not contain tubular complexes, but only a few thylakoids partly surrounding the newly formed starch grains.     

Ultrastructure of the main excretory duct epithelium of the female mouse submandibular gland with special reference to sexual dimorphism

The fine structure of the main excretory duct epithelium (MEDE) of female mouse submandibular gland was investigated by scanning and transmission electron microscopy and the results compared with the previously established structure of male mouse MEDE. A comparative analysis of the subepithelial capillaries of both sexes was also performed. In this pseudostratified epithelium, principal cell-types were observed: types-I,-II,-III and basal cells. This differed significantly from male MEDE, where type-II and-III are absent and type-I cells are the most numerous. The latter cell-type had abundant mitochondria, a few lipid-containing granules, lysosomes in the infra-nuclear cytoplasm and well-developed basal infoldings. These cells were also characterized by abundant glycogen granules throughout the cytoplasm, many profiles of strands of smooth endoplasmic reticulum in the apical region, and lysosomes in the infra-nuclear region. Type-II cells were the second most numerous. Their most characteristic features were the presence of tubular vesicles which appeared to be invaginated from the plasma membrane, RER, SER, free ribosomes, a few peroxisomes with nucleoids, and primary lysosomes in extremely light cytoplasm. They had many mitochondria throughout the cytoplasm, except in the apical region, a few lipid-containing granules and no basal infoldings. Type-III cells were very few and were characterized by well developed basal infoldings, abundant free ribosomes, RER, SER, vesicles containing moderately dense material, and many lipid-containing granules. They also had many mitochondria throughout the cytoplasm, except apically. Basal cells had a large nucleus and the cytoplasm had few organelles. In the male continuous capillaries predominated in the subepithelial network, and capillary density per 200 m of epithelium (3.76±1.54) was lower than in the female, as was the number of fenestrae per 10 m of available endothelium (4.46±1.71). In the female, fenestrated capillaries predominated, and the capillary density per 200 m of epithelium was 6.76 (±1.54), and the number of fenestrae per 10 m of available endothelium was 4.91 (±1.77).     

Chromaffin,small granule-containing and ganglion cells in the adrenal gland of reptiles

Chromaffin, small granule-containing (SGC)-cells, neurons and the innervation of these cells was studied in the adrenal gland of three species of reptiles (Testudo graeca, Lacerta dugesi, Natrix natrix). 1. After fixation with glutaraldehyde and osmium-tetroxide adrenaline (A)- and noradrenaline (NA)-storing cells can be distinguished by means of the different electron density of their granules: A-granules are moderately electron-dense, while NA-granules show a core of high electron density. The unusually high electron density of a few A-granules in Testudo occasionally required viewing of unstained sections which facilitated the discrimination of the two cell types in this species. In all species studied NA-granules display a remarkable polymorphism which is most pronounced in the tortoise. In this species A-granules are polymorphic, too. Both types of granules show wide variations in size, which are particularly great in the tortoise. This species also exhibits the largest average sizes for A-granules (285 nm), and NA-granules (354 nm). The corresponding parameters for Lacerta and Natrix, are 255 and 179 nm for A- and 323 and 304 nm for NA-granules, respectively. The rough ER in A-cells of the tortoise regularly occurs in the form of circular dilations ('ergastosomes', Kanerva and Hervonen, 1973). Mitochondria sometimes contain longitudinal cristae with a crystalloid internal pattern. Large dense bodies which incorporate granules are abundant in NA-cells. Smaller dense bodies containing a few dense patches and membranes are present in both A- and NA-cells. Intermediate stages between dense bodies and what appear to be A- or NA-granules (if the latter have lost some of their amine-content) are frequently observed.     

Endocytosis of native and cationized ferritin by intralobular duct cells of the rat parotid gland

Summary The ability of the intralobular ducts of the rat parotid gland to take up protein from the lumen was examined after retrograde infusion of native and cationized ferritin. At high concentrations (3–10 mg/ml), cells of both intercalated- and striated ducts avidly internalized the tracers. No differences were noted in the mode of uptake or fate of native or cationized ferritin. Large, apical ferritin-containing vacuoles up to 5 m in size were present in cells of the intercalated ducts after infusion for 15 min. Small, smooth-surfaced spherical or flattened vesicles and tubules containing ferritin were also observed, often in association with the large vacuoles. Ferritin uptake increased with increasing infusion time, up to 1 h. Uptake by the striated ducts was less consistent than by the intercalated ducts, and occurred mainly in small vesicles and tubules. Secondary lysosomes became labeled with ferritin in both cell types. Ferritin was not observed in the Golgi saccules, nor was it discharged from the cells at the basolateral surfaces. At low concentrations (0.3–1 mg/ml), uptake was reduced, especially by cells of intercalated ducts, and differences were noted in the behavior of the two tracers. Cationized ferritin was internalized mainly into vesicles and tubules of cells of striated ducts; little uptake of native ferritin occurred at low concentrations. These results demonstrate that the ductal cells of the salivary glands are capable of luminal endocytosis of foreign proteins. They also suggest that in addition to modifying the primary saliva by electrolyte reabsorption and secretion, and secretion of various glycoproteins, the ductal cells are able to reabsorb proteins secreted by the acinar cells.     

Ultrastructural study of the bacillary, granular and mucoid proboscidial gland cells of Riseriellus occultus (Nemertini, Heteronemertini)

Junoy, J., Montalvo, S., Roldán, C. and García‐Corrales, P. 2000. Ultrastructural study of the bacillary, granular and mucoid proboscidial gland cells of Riseriellus occultus (Nemertini, Heteronemertini). — Acta Zoologica (Stockholm) 81 : 235–242. The ultrastructure of six types (G₅‐G₁₀) of proboscidial gland cells whose cell necks emerge independently on the epithelium surface is analysed and compared with data from other nemerteans. These types differ in cytological features, as well as in the morphology of their respective secretory granules. Secretory granules of the types G₅ and G₆ have a bacillary shape, and differ from each other based on their contents and dimensions. Secretory granules of the types G₇ and G₈ are spherical to ovoid; type G₈ gland cells are monociliated, and their secretory granules contain a paracrystalline material. Types G₉ and G₁₀ gland cells are typically goblet‐shaped; secretory granules in the type G₉ have a spherical shape, contain a homogeneous electron dense material and maintain their individuality, whereas those of the G₁₀ type are elongate and have fibrillar contents, showing a tendency to fuse before they are extruded. The mucus sheet of the proboscis is responsible for lubrication of its epithelial surface. Secretion products of type G₁₀ gland cells form the background substance of this mucus, and those of the G₅ type confer stickiness to it. Type G₉ gland cells could provide the toxic component to the mucus, and type G₇ and G₈ gland cells could be concerned with the production of enzymatic secretions.     

Ethnic differences in the expression of blood group antigens in the salivary gland secretory cells from German and Japanese non-secretor individuals

Type 1 ABO blood group antigens (peripheral core structure: Gal1-3GlcNAc1-R) are expressed mainly in endodermally-derived tissues, but are not synthesized in mesodermally-derived tissues. In the former tissues, H type 1 antigen is generated largely by -2-l-fucosyltransferase encoded by secretor (Se) gene and acting on the terminal galactose of the type 1 precursor chain. This theory has been generally accepted, and it seems that the expression of ABO blood group antigens is absent, or expressed at a low level, in these tissues from non-secretor individuals. In this immunohistochemical study on the secretory cells of salivary glands, we found ethnic difference between German and Japanese non-secretor individuals in the expression of blood group antigens: i.e. the expression of the type 1 blood group antigens is present in these cells from Japanese non-secretor individuals but absent from German. A possible explanation is that another -2-l-fucosyltransferase, independent of the secretor gene, is present in Japanese non-secretor individuals.     

Modulations of the epithelial phenotype and functional activity of cultured bovine tracheal gland cells: dependence on the culture medium and passage number.

Bovine tracheal gland (BTG) cells in culture show an epithelial-fibroblastoid transition after several passages. To investigate these BTG cell phenotype changes, we studied the effects of both the culture medium and passage number on the expression of epithelial cytoskeletal proteins and glandular serous cell markers. We also analyzed the intracellular cAMP level in the basal state and after adrenergic stimulation. Three culture media were used: 1) serum-free defined medium (SFDM); 2) medium supplemented with 2% Ultroser G; and 3) medium supplemented with 10% fetal calf serum (FCS). Using immunofluorescence microscopy, we showed that, in the first 4 passages whatever the culture conditions, BTG cells expressed immunoreactivities to cytokeratin filaments and desmoplakins I and II, whereas vimentin filaments were not detected. After four passages, BTG cells cultured in 10% FCS or 2% Ultroser G became progressively fibroblastoid and showed immunoreactivities to both vimentin and cytokeratin intermediate filaments. No immunoreactivity to vimentin filaments was observed on BTG cells cultured in a SFDM. Using biochemical analysis, we showed that basal levels of cAMP in cultured BTG cells and lysozyme secretion by these cells vary according to the culture medium and passage number. It was higher in BTG cells cultured in a SFDM compared to that recovered from cells cultured in medium supplemented with Ultroser G or FCS. Whatever the culture medium, BTG cells responded to stimulation by isoproterenol. However, the results of stimulation in a SFDM were higher than in Ultroser G or FCS supplemented medium. We conclude that the BTG epithelial cell organization and the regulation of biosynthesis of secretory proteins by these cells in culture depend on both the culture medium and passage number.(ABSTRACT TRUNCATED AT 250 WORDS)