Ultrastructure and lectin cytochemistry of the cloacal pelvic glands in the male newt Triturus marmoratus marmoratus

The cloacal organ of Salamandridae species contains four glands: pelvic, dorsal, ventral, and Kingsbury's glands. Pelvic glands have been studied only by light microscopy with conventional methods, and consist of multiple tubular serous glands with a prismatic epithelium which contains numerous PAS positive secretory granules. The present report is an ultrastructural and lectin cytochemistry characterization of the pelvic glands of Triturus marmoratus marmoratus throughout the reproductive cycle. Our methods consisted of conventional electron microscopy, and colloidal-gold lectin cytochemistry of the following lectins: WGA, ConA, LcA, UEA-I, PNA, SBA, and HPA. In the prereproductive period, the glands showed a tall epithelium which consisted of two cell types, dark and clear cells, surrounded by elongated, myoepithelial cells. Both dark and clear cells showed the ultrastructural characteristics of secretory cells, and exhibited many secretory granules in the apical cytoplasm. Areas showing densely packed, degenerating cell organelles--which were not surrounded by membrane--were observed in the dark cells whereas the clear cells showed large heterolysosomes. In the postreproductive period the number of secretory granules decreased, the rough endoplasmic reticulum was less developed, and areas of degenerating organelles were absent. In addition, small basal cells appeared. The results of the lectin histochemistry study were similar in both reproductive periods. In the epithelial cells, the rough endoplasmic reticulum, the Golgi complex, and secretory granules exclusively labeled to ConA. In all cell types, the nuclei reacted to all lectins while the cytosol only reacted to LcA lectin. The ultrastructural and histochemical characteristics of the pelvic glands of T. marmoratus suggest that these glands could be homologous to the mammalian seminal vesicles and prostate.     

Ontogeny of the endocrine cells of the intestine and rectum of sea bass (Dicentrarchus labrax L.): an ultrastructural study

Several endocrine cell types were ultrastructurally characterized during the differentiation of the intestine and rectum of sea bass (Dicentrarchus labrax L.) larvae. Only one cell type (type I) was found in the posterior region of the undifferentiated gut of 5-day-old larvae (phase I). Types V and VI were found in both the intestine and rectum, types II, III and IV in the intestine, and types VII and VIII in the rectum of 9- and 12-day-old larvae (phase II), the rectum alone showing signs of functional differentiation. In phase III larvae, in which both the intestine and rectum were differentiated, types IX, X, XI, XII, XIII, XIV and XV were found in the intestine, only types X, XI and XII being seen in the rectum. Besides these, a new cell type, XVI, was observed in the intestine of 55- and 60-day-old larvae (phase IV), in which the digestive tract was completely differentiated. The endocrine cells appearing in phases I and II showed very scarce secretory granules and the ultrastructural features of undifferentiated cells. Some endocrine cell types in the earliest developmental stages were related to some of those found later. A maturational process of the endocrine cell types paralleled the differentiation of the intestine and rectum, with an apparent increase in the number of secretory granules accompanying organelle development.     

Visceral leishmaniasis: II. Histiocyte ultrastructure in bone marrow associated with low level of parasitaemia and mimicking malignant histiocytosis

The ultrastructural features of histiocytes in the bone marrow (BM) were studied in a febrile, splenomegalic and pancytopenic Sudanese patient who was diagnosed by one of us as visceral leishmaniasis (VL) associated with low level of parasitaemia and mimicking malignant histiocytosis (MH). Serial thick (STS) and ultrathin (SUT) sections showed that the BM was hypercellular and markedly infiltrated by large histiocytes with prominent phagocytosis. A thorough examination of various ST and UT section revealed only a single, typical Leishman-Donovan body. At transmission electron microscopy (TEM) level, two principal types of histiocytic cells were identified: Type I, subdivided into two subtypes, were actively phagocytic histiocytes (PH) with large digestive vacuoles and primary lysosomes; type II were nonphagocytic histiocytes (nPH) with primary lysosomes only. The rate of PH to nPH ws 7:2 in plastic STS. The interaction between the PH and ingested cells is described. Both types of cell were morphologically similar to previously described malignant histiocytic cells. However, this study showed a better characterization of PH during VL.     

The ultrastructure of the rabbit submandibular gland

The secretory granules of the terminal and pre-terminal tracts of the rabbit submandibular gland display distinct features, while the cell morphology is quite similar. Among these cells, some smaller ones presenting an highly dilated endoplasmic reticulum have been identified in the pre-terminal tracts. Some hypothesis have been formulated concerning the function of these cells.     

The ultrastructure of endocrine cells in the corpus of the stomach of human fetuses

The ultrastructural development of endocrine cells from the corpus of fetal human stomachs is described. Samples were taken from fetuses ranging in fertilization age from 6-8 to 22 weeks. The identifying features used for the classification of the various types of endocrine cells were their basal locations in the epithelium, the presence and morphology of their characteristic granules and the sizes of the mitochondria. Five types of endocrine cells with specific granules were found:D, EC, ECL, AL and D1. The type and number of endocrine cells increased as development proceeded. The endocrine cells were confined to the epithelium, they did not reach the lumen and they appeared to develop in situ. The D, EC and ECL cells were the most numerous. The fetal endocrine cells resembled morphologically those found in the stomachs of various adult animals. The EC, ECL and D1 cells contained small slender mitochondria with few cristae. Intramitochondrial granules were absent in all the cell types. Agranular electron-lucent cells with small mitochondria were considered to be immature endocine cells. The advanced stage of differentiation observed in these cells suggest that they may be capable of producing and storing biogenic amines and polypeptide hormones. Their possible involvement in the synthesis of serotonin, enteroglucagon and intrinsic factor is discussed.     

Ultrastructural localization of thyrocyte acid phosphatase in the presence of thyrocyte hyperfunction in rats

Acid phosphatase was found by electron microscopy in the lysosomes which appeared in great numbers in the follicular cells of rat hyperplastic thyroid gland. The other types of granules (mature secretory granules and lipids) whose amounts were also greatly increased in cases of functional thyrocyte strain were also nonreactive. The lysosomes were subdivided into three main groups according to distribution of the reaction product: the lysosomes with dense homogeneous deposit and with deposit in the form of densely or loosely packed dark round granules. The lysosome heterogeneity was apparently connected with their different functions also found within the colloid droplets in the form of inclusions of rarely located dark granules. The authors believe such granules to be the result of the merging of the colloid droplets and lysosomes. The acid phosphatase of the latter participated in the hydrolysis of the product of cell secretion with the formation of active substances.     

The ultrastructure of ependymoma: personal experience and the review of the literature

I report here the ultrastructure of 29 ependymal tumors. The ultrastructural pattern was florid and characteristic with a picture dominated by the presence of microlumina, cilia with basal bodies (blepharoplast), microvilli and long, interdigitating intercellular junctions of the zonulae adherentes (adhesive plaque junctions) type. Tumor cells themselves were not particularly peculiar but they formed typical patterns of rosettes (so called mini- or ultrastructural rosettes) cell gatherings around small, electron-lucent lumina which are filled with numerous microvilli. Empty microlumina were rare. The apical and lateral portions of the cells surrounding microlumina were sealed by intercellular junctions which are long, tortuous and clearly different from the zonulae occludentes (tight junctions) of epithelial tumors. Clusters of apparently "redundant" junctions were occasionally visible comprising segments of different lengths. Ependymoma cells contained myriads of 10 nm intermediate filaments (glial filaments), occasionally forming thick bundles, virtually identical to those encountered in astrocytic tumors and forming an ultrastructural correlate for the GFAP immunostaining. The glycogen granules were often remarkably numerous. Numerous cilia, with a typical 9+1 pattern or with a distorted pattern were frequently observed in longitudinal or cross-sections.     

Morphology and histochemistry of the mandibular gland of the Australian brush-tail possum Trichosurus vulpecula (Marsupialia)

A histological, ultrastructural and histochemical study of the mandibular gland of the Australian possum Trichosurus vulpecula revealed essentially similar features as those described earlier for the mandibular gland of the taxonomically relatively unrelated American opossum Didelphis. The secretory endpieces consist of a branched tubular part, composed of serous cells whose secretion granules contain neutral glycoproteins, and terminal acini, consisting of seromucous cells containing small amounts of sialomucins. Relatively short intercalated ducts lead to striated ducts of variable ultrastructural appearance. The striated ducts run in bundles in the center of each sublobule of the gland. The possible functional significance of the abundance and variability in ultrastructure of the striated ducts is discussed.     

Influence of fasting and stimulation on the rat gastric endocrine cells. Histochemical and ultrastructural study

The ultrastructure and certain cytochemical parameters of endocrine cells of the rat gastric mucosa during 168 h of fasting were investigated. To some of the fasting animals peroral food or alcohol was administered before decapitation. The EC (enterochromaffin cells) the ECL (enterochromaffin-like cells), D1 cells, AL (A-like cells) and G cells were identified by means of electron microscopy. Only the EC, ECL, and G cells could be identified by means of light microscopy by an adequate histochemical technique. The ultrastructural picture of the ECL and of the EC cells did not change markedly during the fasting. In the D1 cells there occurred an agglomeration of secretory granules. Some of them disintegrated and disappeared. In the AL cells an agglomeration of granules during the fasting was also observed. Granules engulfed in lysosomes were often found. The participation of lysosomes in the degradation of granules during the fasting was more marked in the AL cells than in the G cells. The participation of lysosomes was questionable in the EC and D1 cells, and in the ECL cells no lysosomes were observed. In contradistinction to the G cells of the non-fasting animals, where more than one half of the gastrin granules were "empty", the G cells during the fasting were filled with agglomerated dense granules and contained lysosomes with fragments of engulfed secretory granules. Following the administration of food (Larsen's diet) 3 h before sacrificing the dissolution of the content of granules with well preserved membranes was observed (emiocytosis did not take place). The administration of food did not lead to changes in the ultrastructural appearance of the EC cells. The peroral administration of alcohol did not lead to any changes in the ultrastructural appearance of the AL and G cells.     

Fine structural changes of the porcine uterine tube epithelium during early and late pregnancy

Ultrastructural changes in the uterine tube (oviduct) of pregnant gilts have been investigated with special reference to the ciliated, secretory, and stromal cells. Tissue from the uterine tube ampulla and infundibulum was taken from 18 gilts at different stages of gestation (days 31, 36, 101, 102, 107, 110, and 112). Cilia were present throughout pregnancy, and deciliation was not apparent at any stage of gestation. The low epithelium of the uterine tube appeared similar to that of the luteal phase of the estrous cycle when corpora lutea were full grown. Prominent features at end of the gestation were numerous fibrous granules and basal bodies, indicating active formation of ciliary precursor organelles. Fibrogranular aggregates were also present in association with the basal bodies. In addition, numerous polyribosomes, mitochondria, and microtubules were encountered in the cytoplasm of ciliated cells at end of the gestation. The appearance of electron-opaque, fibrous granules during late pregnancy probably could be correlated with increasing endogenous levels of plasma estrogen. Intimate morphologic association between fibrous granules and basal bodies indicate that fibrous granules might provide precursor material for the development of cilia or rootlets. Characteristics ultrastructural changes observed in secretory cells during the estrous cycle were not discernible in secretory cells during pregnancy. The secretory cells appeared similar to those of the luteal phase of the estrous cycle. The apocrine secretory cells contained prominent, apical, cytoplasmic projections; pinching-off process of these protrusions was frequently observed during early and term gestation. Extruded nuclei along with other cytoplasmic organelles were also present, lying free in the tubal lumen. The endoplasmic reticulum was predominantly tubular in form. Synthesis, storage, and release of secretory granules were not apparent at early or late pregnancy. It is suggested that progesterone might have an inhibitory effect on the synthesis, storage, and release of secretory granules. Ultrastructural changes in stromal cells were not apparent at any stage of gestation. The stromal cells appeared similar to that of the luteal phase of the estrous cycle.     

Comparative ultrastructure of vallate, foliate and fungiform taste buds of golden Syrian hamster

A fine-structure study of the hamster fungiform, foliate and vallate taste buds was undertaken for comparative purposes. All three taste bud types shared in common composition of the dark cells, light cells, basal cells, nerve fibers and nerve endings and undifferentiated peripheral cells, but morphological difference existed among them. The foliate and vallate taste buds were quite similar in their ultrastructural morphology. Their dark cells displayed long apical necks, long apical microvilli, apical osmiophilic secretory granules and an abundant rough endoplasmic reticulum. The dark cells of the fungiform taste buds, however, showed no neck formation and lacked apical osmiophilic granules. They had short apical microvilli and relatively scant rough endoplasmic reticulum. There was no difference in the fine structure features of the light cells, basal cells and neural elements of different types of taste buds. Both light and dark cells were much more readily distinguishable in foliate and vallate buds than in fungiform buds at both light-and electron-microscopic levels. Foliate and vallate buds demonstrated homogeneous dense substance within the taste pores while fungiform pores were frequently empty. It is speculated that the differences in taste bud morphology may be due to their different lingual locations and/or may be a reflection of the differences in the inductive influences from different nerves. Furthermore, structural differences may be responsible for varying thresholds to different taste modalities.     

Effects of secretory nerve stimulation on acid phosphatase and peroxidase in submandibular saliva and acini in cats

The effects of parasympathetic or sympathetic nerve stimulation either alone, or in combination, on the acid phosphatase-containing central acinar cells and the peroxidase-containing demilunar cells of the cat submandibular salivary gland have been investigated by histochemical and cytochemical techniques. The results obtained with these techniques were correlated with biochemical assays for both enzymes in the saliva secreted. The results indicate that, although both sets of nerves probably affect both sets of cells, the predominant secretory effect of parasympathetic stimulation is on the central cells and, conversely, the predominant secretory effect of sympathetic stimulation is on the demilunes. Sympathetic stimulation appeared also to have initiated synthesis of peroxidase in the demilunar cells, especially when it was superimposed upon parasympathetic stimulation.     

Genome multiplication mechanisms in the development of albumen gland polyploid cells of Succinea lauta (Gastropoda:Pulmonata). V. Polyploidizing mitosis and endomitosis

Mechanisms of polyploidization of albumen gland cells of S. lauta have been studied using squash preparations in combination with autoradiography of DNA synthesis. During the growth period of the gland normal and abnormal (polyploidizing) mitoses were noticed in addition to endomitoses. Diploid foot cells and poorly differentiated secretory cells are reproduced by the normal mitosis (2c : 2c). These poorly differentiated secretory cells go then through polyploidization by different means. Tetraploid nuclei are substantially formed by abnormal mitoses (4c), which take 40-60% of all mitoses. Metaphase blocks (C-mitosis type) dominate among them; anaphase blocks and their sequences (bladed nuclei) occur rarely. The abnormal polyploid series 3c-6c ... and hypoploid (2c, 3c) mitoses originate from asymmetric mitoses (1c : 3c). Singular nuclei are defined as endomitotic even in the first cycles (4c, 6c). The second and following cycles of the main polyploid nuclei series ... 8c-16c-32c and of alternative series ... 12c-24c pass through endomitosis of classical (insect) type (Geitler, 1939). Unlabelled endomitoses were observed in an hour, while labeled ones were seen in 8-24 hours after 3H-thymidine injection. So, endomitosis can be defined as a real phase of the cell cycle. Integrity of the nuclear envelope as the formal feature of endomitosis was proven at the ultrastructural level (Anisimov, 199). It is emphasized that the normal mitosis is followed by an abnormal polyploidizing mitosis and then by endomitosis. These processes could be considered as compatible and successive polyploidization mechanisms. This conclusion is in agreement with dynamics and speed of DNA and RNA synthesis during cell polyploidization and differentiation (Anisimov, 1988, 1994b, 1995b).     

Endocrine cells in the human fetal small intestine

In this report we describe the time of appearance and ultrastructural features of enteroendocrine (EECs) in the human fetal small intestine (SB) between 9 and 22 weeks gestation. Thirteen distinctive EECs were identified in fetal SB. Two of these, not found in normal adult SB, appeared within the stratified epithelium of the proximal SB at 9--10 weeks. They were arbitrarily termed "primitive" and "precursor" cells. As in all fetal EECs, the pale cytoplasm of the "primitive" cell contains a distinctive population of secretory granules (SGs). Primitive cell SGs average 200-330 nm; some have dense cores with lucent halos while others are filled with a homogeneous dense or flocculent material. The SGs of the "precursor" cells are larger, averaging up to 1 micron in diameter and their contents vary in electron density. A third group of cells not described in normal adult SB was arbitrarily termed "transitional" cells. These have two populations of SGs; one resembles the SGs of the "precursor" cells, and the other resembles the SGs of some of the specific adult type EECs. Transitional EC, S, I and G cells are seen. In addition, mature appearing EC, S, G, I, L, D, and D1 cells were identified by 12 weeks of gestation. The "primitive", "precursor", and "transitional" cells may represent sequential developmental precursors of adult type EECs.     

The ultrastructure of the sheep parotid gland

The sheep parotid is a compound tubular gland; its ultrastructure reflects the function of this gland to secrete large amounts of fluid with very little protein. The cells of the secretory tubules possess extensively folded lateral plasma membranes and a fairly large number of mitochondria. Rapid equilibration of water across the epithelium is assured by the close proximity over large areas of intercellular spaces and the wide secretion canaliculi. Numerous long microvilli extend into the latter. Although secretion granules may be quite numerous, there is evidence that many of these granules are not discharged but undergo degradation by lysosomal enzymes. The intercalated ducts are often dilated but excessive distension is probably prevented by bundles of microfilaments in their epithelial cells.     

Calcitonin gene-related peptide- and substance P-like-immunoreactive innervation of the anterior pituitary in the rat

In our previous studies substantial amounts of substance P- and calcitonin gene-related peptide-like-immunoreactive nerve fibers have been identified in the anterior pituitary of the monkey and the dog. They were found to be in close proximity to the gland cells, even making synaptic contacts with some types of the gland cells. The present study investigated in detail the calcitonin gene-related peptide- and substance P-like immunoreactivities of the anterior pituitary in the rat. Though the immunoreactive fibers were not as abundant as in the anterior pituitary of the monkey and the dog, they still appeared in notable amounts. The calcitonin gene-related peptide- and substance P-like-immunoreactive nerve fibers occurred mostly as thin, tortuous, and densely varicose fibers, weaving among the gland cells. They are widely distributed, more in the central part of the gland. Double-immunostaining proved nearly complete co-localization of these two peptides in the nerve fibers. It is hypothesized that the anterior pituitary can be regulated by direct neural factors as well as humoral factors.     

A histological and ultrastructural study of the cells of the mantle edge of a marine bivalve, Cerastoderma edule

The cells of the mantle edge of Cerastoderma edule are described after light and electron microscopical observations. Histochemical tests for calcium in the mantle edge and digestive gland (Dahl, 1952; McGee-Russell, 1958) and analytical electron microscopy of mantle edge of C. edule both failed to show calcium. Similar results were obtained for Mytilus edulis and Chlamys opercularis. However, calcium was detected in the digestive gland of the terrestrial gastropod Helix aspersa. The outer secretory fold of the mantle edge is composed of tall columnar cells. These cells have highly convoluted lateral cell membranes with which many mitochondria are closely associated. These features are indicative of an ion pump which could move calcium from the mantle space to the extrapallial cavity (compare with Bubel's findings, 1973b). There are many features of the cells lining the periostracal groove of C. edule that have not been reported previously (e.g. Bubel, 1973b) and which are now discussed. The periostracal sheet arises within a line of basal cells in the fundus of the periostracal groove. Within these cells the periostracum in section has a spiral form. It is suggested that the newly formed periostracum adheres to the microvillous border through secretions produced from the middle fold cells lining the groove. During its passage along the groove the periostracum is gradually thickened by secretions from the outer fold cells.     

Characterization of cytoplasmic secretory granules (PSG), in prostatic epithelium and their transformation-induced loss in dysplasia and adenocarcinoma

Cytoplasmic clarity is a histological feature of normal prostatic secretory cells, but in this study, tissue fixation in strong (>2.5%) glutaraldehyde dramatically altered cytological staining. Secretory cytoplasm appeared red and granular on routine stains because of myriad intensely staining eosinophilic granules (PSG). Immunostaining for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) showed their exclusive localization to the PSG. Electron microscopy confirmed these findings and also showed that after fixation in many agents, including formaldehyde, PSG appeared empty, accounting for the artefactual "clear cell" appearance on light microscopy. PSG were most densely concentrated apically in a bud-shaped luminal compartment in which cytokeratin was selectively absent. Normal exocrine secretion was visualized as detachment of apocrine buds or their in situ disintegration. Distinctively in dysplasia and almost all carcinomas, PSG were rare to absent, and proteases were free in the cytoplasm, often concentrated beneath the apical membrane. The apocrine compartment was absent, with no observed secretory mechanism. Tumor cells had dark amphiphilic cytoplasm after all fixatives. This provided a reliable method of distinguishing malignant from benign glands in tissues fixed in strong glutaraldehyde. Clear cell carcinomas, whose cytoplasm mimicked routinely fixed normal secretory cells, surprisingly had almost no PSG. Instead, their "granules" were lipid-filled vacuoles reflecting a secretory pathway not seen in normal cells, dysplasia, or the common "dark cell" carcinomas. These observations may define two distinctive biological pathways of prostate cancer evolution and may facilitate diagnostic decisions on needle biopsy samples.     

Studies on the mechanism of embryo implantation

Implantation is a complex process accomplished by synchronization and interactions between embryo and endometrium by local exchange of signals including a number of cytokines and growth factors and direct cell-cell and cell-matrix contact. However, the research in early events of human implantation is still in its infancy. This presentation comprises the results of our attempts to investigate the mechanisms of human implantation process in its early stage by cell-biological method, including establishment of experimental implantation model in vitro. 1. Human trophoblast of early stage of gestation showed active cell locomotion, active endocytosis, and invasion of endometrial cell monolayer in mixed cultures. Trophoblast invasion was later arrested by transformed endometrial cells similar to decidual cells in vivo. These results appeared to indicate the interactions between trophoblast and endometrial cells in implantation. 2. Coculture system of rabbit preimplantation blastocyst and endometrial epithelium reformed from isolated endometrial epithelial cells on basement membrane matrix (Matrigel) simulated the in vivo rabbit implantation processes. This coculture system may provide a useful experimental implantation model. 3. A human trophoblast cell line was established from chorionic tissues of normal early pregnancy. These cells were cytotrophoblast-like morphology and endocrine functions. They formed the villous structures similar to those in vivo in culture on Matrigel and invasion of Matrigel was observed. These indicated the extracellular matrix may affect the morphology and function of invading trophoblast in implantation site. 4. Human endometrial epithelial single cells were cultured on Matrigel. Reconstruction of gland followed by epithelium formation quite similar to in vivo structures by migration and proliferation of isolated cells was demonstrated. Height of gland was promoted by estrogen and initiation of epithelization was upregulated by platelet-derived growth factors. This system revealed the extracellular matrix regulated morphogenesis of endometrial epithelium in vivo and is an essential substrate in experimental implantation model of endometrial epithelium. 5. Parallel cultures of endometrial epithelial cells on Matrigel were carried out with the IVF. ET patients to evaluate the endometrial morphology at time of ET. Endometrial cultures were initiated in previous cycles on Matrigel and the sera of patients were added to her own cultures from 1st day of IVF treatment cycle. Evaluation of reformed epithelium revealed the apparently unsuitable morphology for implantation in group of patients who eventually failed in pregnancy. This system may provide a useful measures in evaluation of endometrial receptivity and modality of treatment for endometrial aberrations. 6. Cyclic changes of extracellular matrix components in endometrium were investigated. Collagen I, III, IV, V were immunohistochemically estimated. Relative levels of all types of collagen except for collagen V declined at early secretory phase. In rodents, not only collagen but also laminin and fibronectin levels declined at early secretory phase. These changes may facilitate trophoblast invasion of endometrium. Collagen V distributed in myometrial surface was found to consist of subunit (alpha 1)2 alpha 2 and trophoblast growth was inhibited on substrate of alpha 1 subunit. Collagen V in myometrial surface may have a role in blocking trophoblast invasion. 7. HGF (hepatocyte growth factor) mRNA was demonstrated to be expressed during menstruation and secretory phase in endometrium distinctly and its receptor in endometrial epithelial cells and decidual cells. Positive correlation between plasma HGF levels and ultrasonographic thickness of endometrium was observed at late secretory phase. Recombinant HGF promoted proliferation of endometrial epithelial cells and decidual cells and upregulated initiation of endometrial epithelization of Matrigel.