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PubMed1978

Hemidesmosome-like intercellular specializations in human meningiomas.

Desmosomes, gap junctions and tight junctions are intercellular specializations which have been described in human meningiomas. We report, in five of five meningiomas, including a pulmonary metastasis from a malignant intracranial meningioma, a fourth type of intercellular specialization which is similar to but not identical with the hemidesmosome of the epidermis. Idential intercellular specializations have been described between arachnoidal cells, but not between dural fibroblasts. The presence of the junction in human meningeal neoplasms of diverse histologic pattern supports the origin of the meningioma from arachnoidal epithelium. The consistent occurrence of these structures in a pulmonary metastasis as well as benign intracranial tumors suggests that they may be a useful morphologic marker in the differential diagnosis of putative extracranial metastatic meningiomas.

Aged↗
PubMed1983

Freeze-fracture studies of neonatal mouse incisors.

Differentiation of preodontoblasts to odontoblasts and preameloblasts to ameloblasts during development of the mouse mandibular incisor proceeds in a gradient from the area of the odontogenic organ, where undifferentiated ectomesenchymal and epithelial cells proliferate, toward the incisal tip where mature tooth tissues, dentin and enamel, are present. The freeze-fracture technique has been used in the work presented here to study cell membrane ultrastructure of preodontoblasts and preameloblasts at several stages of differentiation. At early stages of differentiation, cuboidal preameloblasts are joined together distally by numerous gap junctions. Relatively fewer junctions occur elsewhere on the lateral plasma membranes, but gap junctions frequently occur proximally between preameloblasts and stratum intermedium cells. As differentiation proceeds and the cells become columnar, distal and proximal junctions persist. Tight junctions, however, were not observed at any of the stages studied. Intramembrane particle concentration of the lateral preameloblast plasmalemma appears to increase as differentiation proceeds. Odontoblasts are also joined distally by numerous gap junctions which persists through later stages of differentiation. Although odontoblast cell processes were observed to project toward the preameloblast layer, no clear points of cell to cell contact or defined intercellular junctions between the two cell types were observed.

Ameloblasts↗
PubMed1979

Electron microscopic studies on the lamprey meninges.

Lamprey meninges were revealed to consist of four layers. The I (outermost) layer was made up of densely packed collagen fibrils and microfibrils with fibrocytes interposed between them. It contained a vascular network (epidural vascular plexus). The II layer was a cellular layer, consisting of two parts: the outer, membranous and the inner, spongy part. The membranous part was composed of two sheets of type 1 cells which were held together by desmosomes, gap junctions and tight junctions. Continuous basal laminae were found on both the outer and the inner surface of the membranous part. The spongy part consisted of type 2 cells, collagen fibrils and microfibrils, which were loosely arranged. Type 2 cells were joined together with each other, or with type 1 cells, by gap junctions. These cells lacked a basal lamina covering. The III layer was a peculiar thick layer, being mostly occupied by fine filamentous matrix substance with scattered round cells, fibroblasts and microfibrils. The IV (innermost) layer was a thick fibrous one, consisting of two parts: the outer, vascular and the inner, loose part. The former consisted of a vascular network (perineural vascular plexus) and thick bundles of microfibrils which frequently contained a dense filamentous core. The latter was composed of loosely arranged collagen fibrils. The inner part of the I layer (inside the epidural vascular plexus) was considered to correspond to the true dura, and the II and IV layer to the arachnoid and pia in the meninges of higher animals, respectively. Characteristically the subarachnoid space was absent in lamprey meninges, and instead a peuliar "gelatinous" layer, III layer, was present between the II and IV layer.

Animals↗
PubMed1987

Impaired spermatogenesis in the finch hybrid L. castaneothorax by L. punctulata: transmission electron microscopy and genetic analysis.

As part of a project to determine whether there is any correlation between the form of hybrid sterility and the genetic relatedness of the parental species, we studied a male intrageneric hybrid between two finch species (Lonchura castaneothorax X L. punctulata), and compared the ultrastructural basis of hybrid sterility in this species with that reported by Swan [1985] for an intergeneric bird hybrid. In the latter study the sterility appeared to have an autoimmune basis, due to lack of Sertoli-Sertoli tight junctions. In the hybrid examined in the present study, lanthanum tracing showed that the junctions were tight. There was no testicular immune reaction; the parental species were almost identical in chromosomal constitution, having only a small inversion difference on chromosome 5, and only two structural protein differences could be detected through examination of the variation at 38 protein loci. Nevertheless, the hybrid appeared sterile and had the following ultrastructural testicular features. Intercellular bridges where present were usually abnormal in structure; centrioles in a centriole pair were arranged in parallel. Many spermatocytes and spermatids degenerated and were phagocytosed by Sertoli cells. Some spermatids progressed to mature testicular spermatozoa in sperm bundles, but commonly had multiple (2-4) axonemes or disrupted doublets and accessory fibers. The multiple axonemes present in most spermatids inserted separately into the base of the nucleus and the multiple centrioles were capable of organizing separate neck structures. We conclude that these cytological abnormalities were caused by genic effects and discuss why they appeared to be restricted to the germ line.

Animals↗
PubMed1995

Cloning of a thymic stromal cell capable of protecting thymocytes from apoptosis.

A clone of thymic stromal cells, namely 2BH4, was established by primary culture, cellular transfection and limiting dilution. Morphological analysis by transmission electron microscopy revealed that these cells grow as multilayers, producing a well-defined basement membrane to which they attach and frequently form structures similar to hemidesmosomes. The adjoining cells are connected by intercellular junctions, as tight junctions, intermediate junctions, and desmosome-like junctions, as well as interdigitations. Their cytoplasm contains microtubules, strands of actin filaments, and scarce intermediate filaments. Fluorescence microscopy revealed that 2BH4 cells stain with anti-cytokeratin antibodies, the majority of them giving a faint reaction. In addition, they express Thy-1.1, LFA-1, ICAM-1, and the gp23 epithelial antigen, and synthesize laminin. They have a doubling time of 16 hr and are able to bind thymocytes. Thymocytes cultured in the presence of 2BH4 cells are partially protected from both spontaneous and PMA- or dexamethasone-induced apoptosis. This protection is conferred neither by soluble factors normally produced by the 2BH4 cells nor by the sole contact with fixed 2BH4 cells. Rather, thymocytes must interact with metabolically active 2BH4 cells in order to receive the protective signal(s).

Animals↗
PubMed1997

Pseudomonas aeruginosa entry into Caco-2 cells is enhanced in repairing wounded monolayers.

Human respiratory cells participating in the repair of epithelial wounds have been shown to be highly susceptible to Pseudomonas aeruginosa adherence. To ascertain whether such susceptibility is a common feature of different repairing epithelial cells, Caco-2 cell monolayers were chemically injured, reincubated for 48 h to partially repair and exposed to bacteria. Cells edging the wounds that spread and migrate to re-establish cell confluence were called 'repairing cells' while cells far from the wounds were called 'non-repairing cells'. By light microscopy, bacteria were seen to adhere to and to enter into both repairing and non-repairing cells. The percentage of intracellular bacteria in repairing cells was significantly higher than in non-repairing cells. The higher susceptibility of repairing monolayers to bacterial entry was confirmed by the gentamicin exclusion assay. P. aeruginosa entry into Caco-2 cells was greatly enhanced in non-injured confluent monolayers treated with EDTA to disrupt intercellular junctions. As tight junction disfunctions have been described during the wound repair process, we speculate that exposure of basolateral receptors to bacterial ligands may account for the enhancement of P. aeruginosa internalization by repairing monolayers.

Bacterial Adhesion↗
PubMed1994

Sequential changes in intercellular junctions between hepatocytes during the course of acute liver injury and restoration after thioacetamide treatment.

Sequential changes of gap junctions (GJs), tight junctions (TJs) and desmosomes (DSs) between hepatocytes during restorative proliferation were studied in rats after a single intraperitoneal administration of 200 mg/kg thioacetamide (TAA). Antibody against connexin 32 was used to demonstrate GJs; simultaneously the changes in TJs and DSs were studied using antibodies against 7H6 protein and desmoplakins. Propidium iodide and bromodeoxyuridine were used to recognize necrotic and proliferative cells. GJs were evenly distributed in early necrotic hepatocytes at 16 h after TAA treatment, then disappeared from necrotic and surrounding cells at 24 h. At 48 h, GJs had disappeared completely from hepatocytes in whole liver lobules, while many hepatocytes were heavily labelled with BrdU. At 72 h, GJs reappeared, firstly in perinecrotic areas. At 96 h after treatment, when the injured areas had disappeared and restorative proliferation ceased, GJs were distributed evenly throughout the lobules. Immunohistochemical observation of GJs in centrilobular, perinecrotic and periportal areas after TAA-induced hepatic necrosis was confirmed by counting the number of connexin-32-positive spots in the respective areas. TJs and DSs disappeared from necrotic cells at 24 h, but then increased between 24 and 48 h in perinecrotic areas, though the increased intensity of these junctions was more evident at 48 h. At 72 h, localization of TJs and DSs returned to normal. These results suggest that during the course of acute hepatic injury, GJs (cell-cell communication) behave differently from other intercellular junctions.

Animals↗
PubMed1983

The pig mesonephros. II. The proximal tubule: SEM, TEM and freeze-fracture images.

The ultrastructure of the proximal tubule of mature mesonephric nephrons was studied in perfusion-fixed pig embryos of the 41st gestational day. Despite its 8-12 mm long course, the proximal tubule possesses no cytologically different subsegments except its very last cells at the abrupt transition into the distal tubule. The first brush-bordered proximal tubule cells stand considerably within Bowman's capsule, abuting its attenuated cells. In SEM specimens, the average luminal cell diameters are 8 X 13 micron. The cells are 6-11 micron in height with overlying microvilli 2-4 micron long. Lateral faces of perfectly disjoined cells exhibit plate-like interdigitating processes projecting more than 5 micron deep into the neighboring cells. The basal cell face is completely covered with microvilli. The TEM pictures reveal an endocytic apparatus largely matching its metanephric counterpart. Mitochondria account for 23% of the cytoplasm and together with the many basolateral cell membranes indicate a high capacity for energy-dependent transport processes. Small basal lipid droplets represent a species peculiarity. Freeze-fracture specimens show an electrocoupling of the cells by gap junctions. The tight junction, with only 1-2 strands, characterizes a "leaky" epithelium. In most features, this tissue is as mature as its metanephric counterpart.

Animals↗
PubMed1990

Membrane modifications in the course of hepatocyte isolation.

A transmission E/M, scanning E/M and freeze fracture ultrastructural study has been performed on the rat hepatocyte in the course of isolation from the liver parenchyma. The cell submicroscopic aspect indicates a good morpho-functional preservation from the liver perfusion to the final stages of cell isolation. The freeze fracture membrane analysis evidentiates the constant presence of gap junctions and tight junctions, characterized by particular structural alterations, probably due to progressive functional uncoupling. The persistence of these cell differentiations until complete cell isolation may be considered a further morphological expression of the maintenance of the differentiated stage of the hepatocyte. Fragments of membranes from adjacent cells, still adherent to isolated hepatocyte surfaces, can also be occasionally detected by freeze-fracture techniques.

Animals↗
PubMed1983

Morphology of the rat carotid sinus nerve. I. Course, connections, dimensions and ultrastructure.

An analysis of the morphology of the carotid sinus nerve in 39 rats revealed that the nerve emerged from the glossopharyngeal nerve (nerve IX) 0.6-1.0 mm beyond the distal extent of the petrosal ganglion. In 68% of cases the nerve consisted of a single bundle of axons; two bundles were present in the other nerves. Near the rostral pole of the carotid body, the nerve divided into multiple bundles of axons. Most axon bundles entered the carotid body, but some instead joined sympathetic nerves from the superior cervical ganglion. Some of the latter group of sinus nerve axons innervated the wall of the carotid sinus. Other presumptive baroreceptor axons reached the carotid sinus from the sinus nerve by first traversing the carotid body. The sinus nerve had an average length of 2.0 +/- 0.13 mm (mean +/- S.E.M., range = 1.3-2.9 mm, N = 15) and was elliptical in cross-section (major axis = 77 microns, minor axis = 46 microns). The nerve contained an average of 625 axons, 86% of which were unmyelinated. The perineurial sheath that enveloped the circumference of the nerve was comprised of 3 or 4 layers of cells interconnected by gap junctions and tight junctions. Near the carotid body, the perineurium extended into the nerve where it compartmentalized fascicles of axons. The endoneurium consisted mainly of collagenous fibres (mean diameter 40 nm) that were only half the size of those outside the perineurium. Endoneurial connective tissue cells were sparse, being only 5% as numerous as Schwann cells. The one blood vessel in the nerve (mean luminal diameter 12.4 +/- 1.2 microns) usually arose from vessels in nerve IX and terminated in a venule at the rostral surface of the carotid body. Ganglion cells were located in nerve IX near the origin of the sinus nerve and along the length of the sinus nerve itself. All ganglion cells examined were postsynaptic to vesicle-containing nerve terminals, and therefore were presumed to be autonomic rather than sensory. Ganglion cells were more numerous in terminal branches of the sinus nerve on the ventral surface of the carotid body. Most other ganglion cells in the carotid body were located among axons of the ganglioglomerular nerve, although some were associated with a small branch of the vagus nerve. Paraganglia were located within the sinus nerve in 15% of cases, and in another 10% of cases they were found nearby in nerve IX. The paraganglia, which measured approximately 50 X 50 X 100 microns, were innervated by axons that resembled sensory nerves of the carotid body.

Animals↗
PubMed1993

Close apposition among neighbouring axonal endings in a neuroma.

Axons in intact peripheral nerve trunks constitute independent afferent and efferent communication channels. However, when nerves are severed, several different forms of axon-axon cross-excitation develop in association with the injury site. In this study we have examined experimental sciatic nerve-end neuromas in rats with special interest in the compartmentalization of individual axons, and the barriers that separate close neighbours. At postinjury times at which functional coupling is known to occur, neuromas were found to contain many examples of axons in which adjacent membrane faces come into close contact without an intervening Schwann cell process. These occur in bundles containing from two to as many as 30 individual nerve fibres wrapped in a common Schwann cell sheath. The surface area of close apposition between axon pairs ranges up to several tens of micron2. Closely apposed axon profiles may be outgrowing branches of a single parent axon, but anterograde tracer data indicate that many belong to independent neurons. Closely apposed axons are separated from one another, and from associated Schwann cell processes, by a cleft about 130 A wide. No synapses, gap junctions or tight junctions were observed. Extracellular tracer studies using La3+ and Ruthenium Red indicated that the cleft system is patent, permitting the free diffusion of small molecules between the space adjacent to the axolemma and the bulk extracellular compartment. Together, these data provide a structural basis for interfibre interactions based on local electrical current flow (ephaptic crosstalk), as well as coupling mediated by K+ ions and neurotransmitter molecules.

Animals↗
PubMed1986

Ultrastructural investigation of fetal rat brain hemisphere tissue in nonadherent stationary organ culture.

Fetal rat brain fragments grown in nonadherent stationary organ culture for 50 days have been investigated ultrastructurally. Synaptogenesis and myelin formation occurred at the same time as the corresponding time-dependent events in the developing brain in vivo. Intermediate junctions were observed between cellular processes lining a central cavity in the fragments and later associated with astrocytes at the surface. Gap junctions and tight junctions were also present. In some fragments cilia were observed in the central cavity. Subependymal basement membrane labyrinths were observed in all fragments after 10 days in culture. The ultrastructural characteristics and the tissue-like structure in general were preserved for at least 50 days in this tissue culture system. The brain fragments may therefore be a valuable supplement to existing culture methods for nervous tissue.

Animals↗
PubMed1987

Cellular junctions on the free surface of human placental syncytium.

Chorionic villi of human placenta at full term have been studied in thin section. On the free surface of syncytium, cellular junctions such as desmosomes, desmosome-like junctions and tight junctions, but not gap junctions, are present. Theories regarding the formation and functions of the junctions on the syncytial free surface are discussed.

Chorionic Villi↗
PubMed1981

Tissue culture of human epithelial cells from benign colonic tumors.

Human colonic epithelial cells from three classes of benign tumors have been reproducibly cultured free of fibroblasts for 8 wk using a supplemented Medium 199 (M 199S). The cultured colonic cells were identified as epithelial by the presence of junctional complexes (tight junctions, gap junctions, and desmosomes), a brush border on the apical surface, keratin fibrils, and by both a close-packed columnar or cuboidal morphology and the capability to transport water and ions to form hemicysts. Colony formation was initiated by groups of epithelial cells, not by single cells, and was inhibited by cocultivation with either lethally irradiated 3T3 cells or human diploid fibroblasts. Enhancement of epithelial colony formation was observed following culture on nonadherent, "floating" substrates compared with substrates attached directly to the bottom of the culture dish. Replication of epithelial cells in M 199S from the class of benign colonic tumors least prone to malignancy, the tubular, was significantly enhanced by epidermal growth factor (EGF). In contrast, EGF did not stimulate the growth of cells in M 199S from the other classes of benign tumors, the villotubular and the villous, which exhibit more malignant potential. These data imply that premalignant colonic epithelial cells lose responsiveness to growth modulation by EGF as they progress toward frank carcinoma.

Adenoma↗
PubMed2004

Calponin is expressed by Sertoli cells within rat testes and is associated with actin-enriched cytoskeleton.

Within seminiferous tubules, Sertoli cells form three types of actin-filament-containing cell junctions, viz., tight junctions, ectoplasmic specializations, and tubulobulbar complexes. These Sertoli cell junctions are involved in the formation of the blood-testis barrier, germ cell translocation, and the release of spermatozoa. Actin and actin-binding proteins are important for these functions. In the present study, a monoclonal antibody against human smooth-muscle-cell calponin detected a 38-kDa protein in a total protein extract of rat testis. The protein has a molecular weight identical to that of aorta calponin and binds calmodulin. Calponin mRNA was detected in the testis and cultured rat Sertoli cells by the reverse transcription/polymerase chain reaction method. Thus, the 38-kDa protein present in the testes is a basic isoform of calponin. In adult rats (70 days of age or older), calponin was detected within seminiferous tubules at sites of Sertoli cell junctions. Cultured Sertoli cells express calponin, whereas cultured spermatogonia do not. When rat testicular tissue was fractionated into cytosol, membrane, and cytoskeleton fractions, calponin was detected in all three fractions. Calcium, potassium, and okadaic acid reduced the amount of calponin associated with the cytoskeleton. These data suggest that Sertoli cells express calponin. The function of calponin within the testis is as yet unknown.

Actins↗
PubMed1985

Intercellular junctions in human tooth-pulp cells in culture in vitro revealed by freeze-fracture, lanthanum impregnation and filipin treatment.

Three kinds of intercellular junctions were detected between human dental pulp cells in explant culture with electron microscopy included filipin detection for cholesterol; desmosome-like junctions observed on ultrathin sections probably contribute to the cohesiveness between cells in culture. Gap junctions, responsible for intercellular communication, exhibited two morphologies on freeze-fracture replicas: a conventional arrangement of their intramembranous particles and a crystalline array corresponding to the formation stage of junctions. Primitive tight junctions were detected on freeze-fracture replicas but not on ultrathin sections. It is likely that they contribute to the cell-to-cell adhesion under culture conditions.

Cells, Cultured↗
PubMed1976

Isolation of the basal and lateral plasma membranes of rat kidney tubule cells.

A method was developed to isolate renal basolateral membranes from cortical kidney tubule cells of single rats. The isolated membrane fraction was characterized by the measurement of marker enzyme activities and by electron microscopy. 1. After centrifugation of crude plasma membranes on a discontinuous sucrose density gradient the basolateral membranes accumulated at a sucrose density of p= 1.14-1.15 g/ml. The yield was 147 mug membrane protein/g kidney wet weight. Protein recovery was 0.1%. 2. (Na+ + K+)-ATPase was enriched 22-fold from the homogenate. The recovery was 2.6%. The (Na+ + K+)/Mg2+-ATPase ratio was 4.1. 3. The contamination by brush borders was small. Alkaline phosphatase was 1.6-fold enriched and 0.2% was recovered. Aminopeptidase was 1-fold enriched with a recovery of 0.1%. The contamination by mitochondria, lysosomes and endoplasmic reticulum was negligible. 4. In electron micrographs the basolateral membranes showed a typical triple layered profile and were characterized by the presence of junctional complexes, gap junctions or tight junctions.

Adenosine Triphosphatases↗
PubMed1994

Ultrastructure and permeability of endothelial cells in branched regions of rat arteries.

The ultrastructure and the permeability to macromolecules of the endothelia in the branched and unbranched regions of the arteries were compared using two different age groups (3 and 12 months) of rats. In the aortic arch, the endothelial cells were longer and thinner and contained fewer intracytoplasmic vesicles than those observed in the unbranched regions of aorta. Quantitative study revealed that the volume density of intracytoplasmic vesicles in the branched regions of aortic arch in 3-month-old rats was significantly (P < 0.01) lower than the density value in the unbranched regions of aorta. The volume densities of vesicles in both regions of the aorta were lower than those in the carotid artery. There was an apparent increase in the frequency of the simple type of interendothelial contacts and a decrease in the complex type in the branched regions as compared with those in the unbranched regions of aorta and carotid artery. In addition to the normal interendothelial contacts, several open junctions with increasing width (25-300 nm) were identified in the branched regions of aortic arch and the bifurcations of carotid artery. For rats at the age of 12 months, local areas of the subendothelial space were expanded. Basal lamina-like and electron-dense materials were accumulated in the subendothelium. The volume densities of vesicles in the aortic endothelia were significantly (P < 0.01) increased as compared with those in the 3-month-old group. The volume density of vesicles in the aortic arch was again significantly (P < 0.01) lower than that in the unbranched regions of aorta. Furthermore, the frequency of the simple type of intercellular contacts was increased, whereas that of the complex type was decreased in both regions of aorta. With regard to the junctional complexes, the frequencies of gap junctions and tight junctions were increased and the junctionless intercellular contacts were decreased compared with those of the 3-month-old group.

Aging↗