Spontaneous hyperplastic and metaplastic duct epithelium in the sublingual salivary glands of Wistar rats

Focal epithelial hyperplasia and metaplasia were observed in the intralobular ducts of the sublingual salivary glands of 302 out of 1142 Wistar rats. These rats (aged 16-145 weeks) served as controls in toxicological experiments. The ductal changes varied from small groups of irregular cylindrical cells with basophilic cytoplasm and vesicular nuclei, with often prominent nucleoli, to large areas of non-keratinized stratified squamous epithelium with marked mitotic activity and necrotic superficial cells being released in the lumen. Mononuclear inflammatory cells were often present in fibrous tissue surrounding altered ducts. Within certain limits the degree and incidence of the changes increased with increasing age; their aetiology is unknown.     

大鼠颌下腺促性腺激素释放激素及其mRNA的免疫组织化学与原位杂交研究

用免疫组织化学及原位杂交法,研究了促性腺激素释放激素及其ｍＲＮＡ在大鼠颌下腺的分布。结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞,各级导管的上皮细胞及副交感神经节细胞均呈促性腺激素释放激素免疫反应阳性,阳性反应物质分布在胞质,胞核呈阴性反应。颌下腺的浆液性腺泡上皮细胞,各级导管上皮细胞同样被检测到很强的促性腺激素释放激素ｍＲＮＡ杂交信号。以上结果提示,大鼠颌下腺能自身合成促性腺激素释放激素,促性腺激素释放激素对消化功能可能有重要调节作用。     

Ultrastructure of genital system ducts of Diphyllobothrium latum (Cestoda:Pseudophyllidae): the ducts of the female reproductive system

The components of the female reproductive system of Diphyllobothrium latum, including ovary, ovicapt, oviduct, vitelline ducts, vitelline reservoir, vagina, seminal receptacle, ootype with unicellular Mehlis's gland, ootype-uterine duct and uterus were observed with the electron microscope. The epithelium of the female reproductive system ducts consists of a nucleate syncytial layer. Structural differences in apical surface of the ducts, the number of nuclei and organoids in syncytial layer as well as the number of underlaid muscles were revealed. The regional differentiations of the uterus wall were registered. The middle and distal region of uterus was covered with microtriches. The filamentous microtriches were observed in apical surface of vagina. The epithelium of seminal receptacle and distal region of uterus were underlaided by the powerful muscle layers. The fertilization canal was revealed. It was shown that the formation of egg shells implemented by the deposit of vitelline globules in their surface in the ootype-uterine duct. Structural and functional differences of different parts of female apparatus in various groups of cestodes are conditioned by species biology.     

The differentiation of oval cells into hepatocytes during induced hepatic carcinogenesis in mice. An electron microscopic study

An electron microscopic study of murine oval cells, induced by a single injection of genotoxic agent dipin and by a partial hepatectomy, has shown that their ultrastructure and direction of differentiation depend on localization in the liver lobule. Oval cells around portal tracts go through three stages of development: low differentiated cells 4.40 +/- 0.51 mu in diameter with ovoid nuclei 3.43 +/- 0.44 mu, intermediate cells, and young hepatocytes. They form common ducts surrounded by a basal lamina, and produce bile canaliculi-like structures and intermediate junctions between them. Another part of the oval cell population is organized similar to the bile duct epithelium. It consists of cells 9.37 +/- 1.1 mu in diameter with nuclei 7.28 +/- 1.16 mu in diameter and form a system of branching and anastomosing ducts widespread along the parenchyma from the portal to the central veins. Our data indicate that the oval cells can differentiate into hepatocytes, and support a hypothesis according to which the cells of terminal bile ductules are liver epithelial stem cells which can differentiate into a hepatocyte or a bile duct cell lineage in periportal microenvironment.     

Immunolocalization of centriole-associated striated rootlets in human submandibular gland cells with and without solitary cilia

Using an antibody specific to striated rootlets, we investigated the immuolocalization of striated rootlets in cells constituting human submandibular glands. Striated rootlets were positively stained in all cell types constituting acini, intercalated ducts, striated ducts, and interlobular ducts, but their shapes were different. The mean lengths of striated rootlets were 1.46 +/- 0.49, 3.15 +/- 1.35 and 3.99 +/- 1.02 microm in acinar secretory cells, myoepithelial cells, and columnar cells of the striated duct, respectively. The rootlets were the longest in columnar cells of the striated duct, in which paired centrioles were located in the apical cytoplasm away from nuclei. These findings suggest that striated rootlets play important roles in the positioning of centrioles in the cell. 2-8% of striated rootlets in myoepithelial cells were associated with solitary cilia, but they were not associated with solitary cilia in acinar cells and columnar cells of the striated duct. These observations suggest that striated rootlets may be associated with centrioles under normal physiological conditions, without formation of solitary cilia.     

Compensatory proliferative response of the rat submandibular salivary gland to unilateral extirpation

The present experiment was conducted in order to identify the progenitor compartment of the submandibular salivary gland (SSG) and to explore the proliferative activity of this gland in response to unilateral extirpation. Left submandibular and retrolingual glands were extirpated in 30 rats (B.W. 200 +/- 12 g). The rats were killed 0, 1, 2, 3, 5 and 7 days after surgery. Five intact rats served as controls. The animals were given intraperitoneal injections of 3HTdR (0.5 microCi/grB. W) 1 h before they were killed. The contralateral SSG's were subjected to routine histological procedures and embedded in glycol methacrylate. Selected sections (2 micron thickness) were processed for autoradiography. In each gland, labelled and unlabelled nuclei were counted in 50 random microscopic fields and sorted according to their parenchymal histomorphological features and "nuclear class" (number of nuclei/cross section/feature). In the control glands the total labelling index (LI) was 0.18%; during acute compensatory stimulation, however, the total LI reached a maximum of 0.86% on day 3 after surgery. suggesting that the SSG, which normally undergoes a slow turnover, is capable of elevated proliferation in response to a stimulus. In both normal and stimulated glands, the LI was higher in the intercalated ducts (1.1%-5.85%) than in the granular ducts (0.17%-0.93%) and acini (0.05%-0.36%). This consistency of LI ratio between the various histomorphological features in the normal and experimental glands indicates that the glandular progenitor compartment is located in the intercalated ducts, which supply cells to both the ductal system and acini.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ultrastructural investigations of the salivary glands in adults of the microtrombidiid mite Platytrombidium fasciatum (C. L. Koch, 1836) (Acariformes: Microtrombidiidae)

The organization of the salivary glands in ad libitum-fed adult females of the microtrombidiid mite Platytrombidium fasciatum (C. L. Koch, 1836) was observed using transmission electron microscopy. In all, four pairs of large simple alveolar salivary glands were determined, which have been named due to their position as posterior, ventral, medial and dorsal. These glands occupy a body cavity behind, around the base and partly inside the gnathosoma. The posterior glands are largest and possess large nuclei with greatly folded nuclear envelope. Secretory granules are electron-light, containing fine granular material and are partly provided with various lamellar inclusions inside the granules. The latter tend to be placed predominantly in the middle parts of the gland around the central (intra-alveolar) cavity. The remaining glands, conversely, are typically filled with tightly packed electron-dense secretory granules, except for the ventral glands, the granules of which may show a compound organization. The nuclei of all these glands occupy a peripheral position and are mostly pressed between the granules. No prominent endoplasmic reticulum or conspicuous Golgi bodies were observed within the salivary glands. The salivary glands are provided with a complex apparatus of the intra-alveolar cavity (acinar lumen) with the excretory duct base provided by a system of branched special cells producing the duct walls. The ventral glands open by separate ducts into the most posterior part of the subcheliceral space. Ducts of the posterior glands immediately fuse with the ducts of the tubular (coxal) glands. The common duct of each side of the body joins with the ducts of the medial and dorsal glands respectively, and opens into the subcheliceral space far anterior to that of the ventral glands.     

Computer ranking of the sequence of appearance of 40 features of the brain and related structures in staged human embryos during the seventh week of development

The sequence of events in the development of the brain in human embryos, already published for stages 8-17, is here continued for stages 18 and 19. With the aid of a computerized bubble-sort algorithm, 58 individual embryos were ranked in ascending order of the features present. The increasing structural complexity provided 40 new features in these two stages. The chief characteristics of stage 18 (approximately 44 postovulatory days) are rapidly growing basal nuclei; appearance of the extraventricular bulge of the cerebellum (flocculus), of the superior cerebellar peduncle, and of follicles in the epiphysis cerebri; and the presence of vomeronasal organ and ganglion, of the bucconasal membrane, and of isolated semicircular ducts. The main features of stage 19 (approximately 48 days) are the cochlear nuclei, the ganglion of the nervus terminalis, nuclei of the prosencephalic septum, the appearance of the subcommissural organ, the presence of villi in the choroid plexuses of the fourth and lateral ventricles, and the stria medullaris thalami.     

THE ULTRASTRUCTURE OF DEVELOPING LATEX DUCTS IN MAMMILLARIA HEYDERI (CACTACEAE)

Electron microscopy was used to investigate early development of latex ducts in Mammillaria heyderi (Cactaceae). Numerous vesicles (secondary vacuoles) form from invaginations of the plasmalemma near sites of wall thinning, from endoplasmic reticulum (ER), and from vesiculate grana of degenerate plastids. Dictyosomes, though they occur in young duct cells, do not seem to be responsible for the formation of vesicles. Cytoplasmic vesicles may contain fibrillar, globular, or crystalline materials, or may be devoid of any type of particulate matter. They may be responsible for storage of numerous laticiferous components. Lysosomal materials could be stored in some vesicles and contribute to the degradation of the protoplast. Some nuclei contain condensed chromatin and are subject to deformation and collapse. Mitochondria and lipid bodies are common in young duct cells but ER is rare. When ducts form in young tissues, plastids in the lumen do not produce starch grains or extensive membranous networks. The plastids eventually degenerate to become a part of latex. If ducts form in older, established tissues having mature plastids, the plastids undergo extreme modification.     

Quantitative analysis of prostatic intraepithelial neoplasia on tissue sections.

The changes in nuclear morphology (karyometry) and DNA content in prostatic intraepithelial neoplasia (PIN) were analyzed on tissue sections. The cases of PIN were subdivided into PIN 1 and PIN 2 based on the degree of proliferation and the anaplasia of the secretory cells lining the ducts and acini. Cases of nodular hyperplasia (NH) and adenocarcinoma were also studied for comparative purposes. Karyometric analysis showed a progression of most values from NH to PIN to carcinoma. The DNA analysis showed a decrease in the frequency of nuclei in the diploid range and an increase in the percentage of nuclei in the other ploidy regions (especially between 2c and 4c and in the tetraploid range) from NH to PIN to carcinoma. Forward stepwise discriminant analysis showed similarities between NH and PIN 1 and between PIN 2 and carcinoma. These findings suggest that the evolution towards adenocarcinoma is characterized by progressive morphologic derangements of the nuclei and by the transformation of the diploid DNA content into a nondiploid one, with the changes taking place at the level of PIN 2.     

Development of the duct system during exocrine pancreas differentiation in the grass snake Natrix natrix (Lepidosauria,Serpentes)

We analyzed the development of the pancreatic ducts in grass snake Natrix natrix L. embryos with special focus on the three‐dimensional (3D)‐structure of the duct network, ultrastructural differentiation of ducts with attention to cell types and lumen formation. Our results indicated that the system of ducts in the embryonic pancreas of the grass snake can be divided into extralobular, intralobular, and intercalated ducts, similarly as in other vertebrate species. However, the pattern of branching was different from that in other vertebrates, which was related to the specific topography of the snake's internal organs. The process of duct remodeling in Natrix embryos began when the duct walls started to change from multilayered to single‐layered and ended together with tube formation. It began in the dorsal pancreatic bud and proceeded toward the caudal direction. The lumen of pancreatic ducts differentiated by cavitation because a population of centrally located cells was cleared through cell death resembling anoikis. During embryonic development in the pancreatic duct walls of the grass snake four types of cells were present, that is, principal, endocrine, goblet, and basal cells, which is different from other vertebrate species. The principal cells were electron‐dense, contained indented nuclei with abundant heterochromatin, microvilli and cilia, and were connected by interdigitations of lateral membranes and junctional complexes. The endocrine cells were electron‐translucent and some of them included endocrine granules. The goblet cells were filled with large granules with nonhomogeneous, moderately electron‐dense material. The basal cells were small, electron‐dense, and did not reach the duct lumen.     

Development, Histochemistry and Ultrastructure of Gum-resin Ducts in Commiphora mukul Engl.

Gum-resin ducts are present in the primary and secondary phloemof Commiphora mukul Engl. The important gum-resin, known commerciallyas ‘guggul’, is secreted and collected in ductswhich develop schizogenously. The duct initials have dense cytoplasm,large nuclei, increased cytoplasmic RNA and proteins. The lumenof newly-formed ducts widens accompanied by anticlinal divisionsand subsequent tangential elongation of epithelial cells. Histochemicaltests reveal that the epithelial cells have apparently largeamounts of proteins, cytoplasmic RNA, and DNA in the nucleus.Lipid globules are also present in these cells. Epithelial cellwalls in contact with the duct are thin and of a loose fibrillarmesh. Microtubules, randomly oriented in the epithelial cellsare always parallel and adjacent to the wall. The cytoplasmis rich in ribosomes, endoplasmic reticulum, mitochondria, plastidsand vacuoles containing osmiophilic substances. At the peripheralregion of the duct, electron-transparent bodies containing densely-stainedmaterial are present close to the tangential wall.     

Localization of c-ERB A proteins in rat liver using monoclonal antibodies

Monoclonal antibodies were raised against the nuclear thyroid hormone receptors encoded by c-ERB A genes and against a purified nuclear receptor fraction. These antibodies recognize the c-ERB A protein in nuclear extracts from rat liver and are able to compete with thyroid hormone in Scatchard analyses. In sections of rat liver they react with all the hepatocyte nuclei as well as with the cells of the hepatic bile ducts. Comparison with another putative T3 receptor antibody, described previously, showed that distinct 57 kD proteins with a different cellular distribution were recognized.     

Smad2/3蛋白及其活化形式在人肾脏中的表达和定位

目的研究Smad2和Smad3蛋白及其活化形式P—Smad2、P—Smad3在人正常肾脏组织中的表达、定位及其意义。方法采用免疫组织化学技术（SP法）检测20例人肾脏中Smad2、Smad3蛋白及P—Smad2、P-Smad3的表达和定位。结果Smad2、Smad3在肾小管、肾小球和集合小管中广泛表达,主要定位于细胞质,其中远端小管曲部呈强阳性;P-Smad2、p-Smad3也在肾脏皮、髓质中广泛分布,主要定位于细胞核,远端小管更多见。结论Smad2、Smad3在正常肾脏中有着活跃的功能。     

Occurrence and Nuclear Localization of cAMP Response Element- Binding Protein in the Post-natal Development of the Rat Submandibular Gland

Cyclic AMP response element-binding protein (CREB) is a 43-kDa polypeptide that binds a cAMP response element located at the 5 promoter region of cAMP regulatory genes. The spatial and temporal distribution of CREB in the post-natal development of the rat submandibular gland was investigated using immunohistochemistry with a specific antibody. At birth, cells of the terminal tubules and ducts in the submandibular gland showed a nuclear CREB immunoreactivity of moderate intensity. At 1–2 weeks after birth, an intense CREB immunoreactivity was localized primarily to acinar cells. When the r352;-adrenergic agonist isoproterenol was administered to 2-week-old rats, a twofold transient increase in the number of immunoreactive acinar cells was induced. Beginning 3 weeks after birth, CREB immunoreactivity shifted from acini to the duct system and showed a clear localization in the cells of the intercalated ducts and distal portions of striated ducts, where the granular convoluted tubule develops after 4 weeks. Immunopositive materials were localized exclusively in the nuclei of both acinar and ductal immunoreactive cells. After the development of the granular convoluted tubules, CREB immunoreactivity was absent in the tubule cells and was gradually reduced in intensity over the entire gland. In order to examine a hypothesis that CREB is involved in the initial differentiation of the granular convoluted tubular cells, testosterone was administered to hypophysectomized adult rats. Whereas the tubular cells of hypophysectomized rats showed a complete regression, and no CREB immunoreactivity was found in any acinar or duct cells, administration of testosterone for a few days induced an intense CREB immunoreactivity in the nuclei of duct cells, followed by their differentiation into the granular convoluted tubular cells. These results suggested that CREB is involved not only in the growth and differentiation of acinar ce lls that are regulated by r352;-adrenergic nerves but also in those of the duct system, and especially in the androgen-regulated differentiation of the granular convoluted tubular cells, during the post-natal development of the rat submandibular gland.     

Ultrastructure of the larval salivary glands of Megaselia scalaris Loew (Diptera,Phoridae)

Each of the paired salivary glands of third instar larvae of the humpbacked fly Megaselia scalaris is a bag-like structure with a short neck region from which a single duct emerges. The two ducts form a common duct that empties into the ventral region of the pharynx near the mouthparts. The wall of the glands and ducts consists of a simple squamous epithelium that rests upon a connective tissue layer. Cells in the neck are less flattened than those found elsewhere. The basal surfaces of the cells are infolded most deeply in the neck and the least in the duct. The apical surfaces of the cells possess microvilli except in the duct where the apices of the cells are covered by a complex extracellular layer. This layer displays circularly arranged folds that accommodate a thread-like supportive structure resembling taenidial threads of tracheae. Elaborate junctional complexes are associated with the lateral surfaces of the cells. Elements of these complexes include a zonula adherens, a series of pleated septate desmosomes, and conventional desmosomes. The cytoplasm of the glandular cells is filled with RER and other organelles normally seen in cells that export proteins and mucosubstances. Secretory material found in the lumens of the glands reacts only moderately with the PAS procedure but more strongly with alcian blue and methods that demonstrate proteins. The nuclei of the glandular cells contain single large nucleoli and polytene chromosomes whose banding is rather indistinct. Treatment with EDTA produces detrimental effects on all of the foregoing ultrastructural features of the glands and ducts.     

Immunohistochemical localization of activated EGF receptor in human eccrine and apocrine sweat glands.

Epidermal growth factor (EGF) is secreted into sweat from secretory cells of human sweat glands. The function of EGF in sweat is poorly understood. The biological function of EGF is exerted by the binding of EGF to the receptor (EGFR) and its activation. Therefore, we immunohistochemically localized the activated form of EGFR in human eccrine and apocrine sweat glands to assess the functional importance of the EGF-EGFR system in human sweat glands. Frozen sections of human skin were stained with a monoclonal antibody (MAb) specific for tyrosine-phosphorylated (activated) EGFR and with an MAb that stains both activated and non-activated EGFR. In the secretory portion of eccrine sweat glands, nuclei of the secretory cells were stained with the anti-activated EGFR MAb. In coiled and straight portions of eccrine sweat ducts, nuclei of luminal and peripheral cells were stained with the antibody specific for activated EGFR. Luminal cell membranes and luminal cytoplasm of inner ductal cells possessed non-activated EGFR. In the secretory portion of apocrine sweat glands, activated EGFRs were present in cytoplasm and nuclei of secretory cells. These data suggest that EGF, already known to be present in the cytoplasm of secretory cells in eccrine and apocrine sweat glands, activates EGFR in the nuclei of secretory cells themselves in an intracrine manner. Because ductal cells do not express EGF, EGF in the sweat secreted from the secretory cells should activate EGFR in the ductal cells in a paracrine manner. (J Histochem Cytochem 49:597-601, 2001)     

Presence of Marteilia sp. (Paramyxea) in the razor clam Solen marginatus (Pennántt, 1777) in Galicia (NW Spain)

Protistan parasites of the genus Marteilia, phylum Paramyxea, cause the molluscs disease named Marteiliosis. Histological observations and transmission electron microscopy revealed the presence of life cycle stages of a Marteilia sp. in the bivalve mollusc Solen marginatus (Solenidae). Parasites occurred in epithelial cells of the digestive ducts and tubules. Early stages (primary cells) presented one or several nuclei while advances stages formed a complex of cells-within-cells (secondary and tertiary cells) culminating in spores. Refringent bodies were present inside the presporangia. This is the first report of a Marteilia sp. in S. marginatus.     

Recurrence of posterior polymorphous corneal dystrophy is caused by the overgrowth of the original diseased host endothelium

Peplomycin (PEP), an anti-tumor antibiotic related structurally to bleomycin, is widely used, especially for squamous cell carcinoma but shows renal toxicity. We prepared monoclonal antibodies (mAbs) against N-(γ-maleimidobutyryloxy)succinimide-conjugated PEP. The mAbs were monospecific for PEP, but did not react with bleomycin and other anticancer antibiotics. The mAbs enabled us to develop an immunocytochemical (ICC) method for detecting the uptake of PEP in the rat kidney. Two hours after a single i.v. administration of PEP, ICC revealed immunostaining for PEP in irregularly shaped cytoplasmic granules of the proximal tubules in which the microvilli were also stained. Also, staining occurred in the distal tubules and collecting ducts, in both of which we observed scattered swollen cells, reminiscent of necrotic cells, in which both the nuclei and cytoplasm reacted strongly with the antibody. Twenty-four hours after injection, PEP in the proximal tubules completely vanished, but yet significant amounts of PEP remained in both the distal tubules and collecting ducts. Distribution patterns of PEP in cells of the kidneys resembled, in some ways, those of our recent ICC studies for an organic cation aminoglycoside antibiotic gentamicin. This ICC suggests that PEP taken up in the proximal tubule cells is localized in the lysosomes, and organic cation transporters and bleomycin hydrolase might be involved in entrance and/or disappearance of PEP in this cell type. Furthermore, the distal tubules and collecting ducts may be the sites readily affected by some chemotherapeutic agents.