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Intercellular junctions in the gill epithelium of the Atlantic hagfish, Myxine glutinosa.

The intramembrane organization of the occluding junctions in the gill epithelium of the Atlantic hagfish, Myxine glutinosa, was studied by means of freeze-fracture electron microscopy. Mitochondria-rich cells, characterized by assemblies of rod-shaped particles in the luminal plasma membrane and by an extensive intracellular amplification of the basolateral plasma membrane, are singly distributed between the pavement cells in the gill epithelium of this marine and stenohaline cyclostome. The occluding junctions between mitochondria-rich cells and pavement cells do not differ from those between adjacent pavement cells, concerning the number of superimposed strands (median 6, range 4-9) and their geometrical organization. These observations suggest that, in contrast to marine teleosts, the paracellular pathway plays a minor role in transepithelial ion movements in the hagfish gill epithelium. The findings are in agreement with the absence of hypoosmoregulatory mechanisms in hagfish, as have been evolved in various marine vertebrates. In addition, small communicating junctions are demonstrated between pavement cells; they possibly serve for a coordinated synthesis and secretion of mucus by the pavement cells.

Animals↗

Germ cell-somatic cell relationships: a comparative study of intercellular junctions during spermatogenesis in selected non-mammalian vertebrates.

Specialized germ cell-somatic cell relationships were surveyed in the testis of species representative of four classes of non-mammalian vertebrates. Desmosome-like junctions were present in all classes studied. In the teleost fish studied (bluegill; Lepomis macrochirus), small, infrequent desmosomes, seen between the spherical cyst cells and spermatocytes, were characterized by poorly represented subsurface densities. In the bullfrog (Rana catesbeiana), similar desmosome-like junctions were found between cyst cell processes and spermatocytes. Reptilian (turtle; Pseudameys scripta) desmosome-like junctions between Sertoli cells and germ cells were heterogeneous and more numerous than those junctions found in fish and amphibians. In general, the reptilian desmosome-like junctions were extensive structures displaying 10 nm filaments associated with the Sertoli cell component of the junctions. Regions within the desmosome where the two plasma membranes converged suggested that gap junctions were a component of the desmosome-like junctions. "Desmosome-gap" junctions persisted in turtle spermatids for sometime after nuclear elongation had commenced. In birds (chicken; Gallus domesticus), "desmosome-gap" junctions, similar to those seen in turtles were described between both spermatocytes and Sertoli cells, and spermatids and Sertoli cells. These junctions were frequently lined by saccules of endoplasmic reticulum. The presence of gap junctions suggest the evolution of mechanisms for somatic cell-germ cell communication although more species should be examined to confirm this hypothesis.

Animals↗

The Drosophila clock gene per affects intercellular junctional communication.

The per locus of Drosophila has been implicated in the control of behavioural rhythms. In fruitfly embryos and larvae per is expressed in salivary glands. Per mutations have striking effects on intercellular communication in salivary glands: gap junction channels are modulated so that their conductance varies inversely with the period of behavioural rhythms in the mutants. A similar effect on junctional communication in the nervous system may explain how per influences behavioural rhythms.

Animals↗

Breaking the connection: displacement of the desmosomal plaque protein desmoplakin from cell-cell interfaces disrupts anchorage of intermediate filament bundles and alters intercellular junction assembly.

The desmosomal plaque protein desmoplakin (DP), located at the juncture between the intermediate filament (IF) network and the cytoplasmic tails of the transmembrane desmosomal cadherins, has been proposed to link IF to the desmosomal plaque. Consistent with this hypothesis, previous studies of individual DP domains indicated that the DP COOH terminus associates with IF networks whereas NH2-terminal sequences govern the association of DP with the desmosomal plaque. Nevertheless, it had not yet been demonstrated that DP is required for attaching IF to the desmosome. To test this proposal directly, we generated A431 cell lines stably expressing DP NH2-terminal polypeptides, which were expected to compete with endogenous DP during desmosome assembly. As these polypeptides lacked the COOH-terminal IF-binding domain, this competition should result in the loss of IF anchorage if DP is required for linking IF to the desmosomal plaque. In such cells, a 70-kD DP NH2-terminal polypeptide (DP-NTP) colocalized at cell-cell interfaces with desmosomal proteins. As predicted, the distribution of endogenous DP was severely perturbed. At cell-cell borders where endogenous DP was undetectable by immunofluorescence, there was a striking absence of attached tonofibrils (IF bundles). Furthermore, DP-NTP assembled into ultrastructurally identifiable junctional structures lacking associated IF bundles. Surprisingly, immunofluorescence and immunogold electron microscopy indicated that adherens junction components were coassembled into these structures along with desmosomal components and DP-NTP. These results indicate that DP is required for anchoring IF networks to desmosomes and furthermore suggest that the DP-IF complex is important for governing the normal spatial segregation of adhesive junction components during their assembly into distinct structures.

Cadherins↗

Structure of intercellular junctions in the endothelium.

Endothelial cell junctions are complex structures formed by transmembrane adhesive molecules linked to a network of cytoplasmic/cytoskeletal proteins. At least three different types of endothelial junctions have been described: tight junctions, gap junctions and adherens junctions. These structures have some features and components in common with epithelium but also some which are specific for endothelium. We still know very little about the pathologic consequences of alterations in the functional behaviour or synthesis of endothelial cell junction proteins. It is possible that pathologies linked to altered endothelial permeability and vascular organization (e.g. hemangiomas, scleroderma, and other types of vasculitis) are associated with structural alterations in endothelial junction organization. In addition, changes in endothelial permeability properties are associated with the early stages of atherosclerosis and many inflammatory diseases.

Capillary Permeability↗

Fine structure of intercellular junctions and blood vessels in medulloblastomas.

Six medulloblastomas were studied by electron microscopy. Two features were found which seem to be constant and essential characteristics of medulloblastoma. First, cell junctions are abundant between tumor cells. These are mostly desmosome-like but other, closer junctions, were also seen. Second, the capillary endothelia contain frequent tubular bodies and other inclusions which may be related to them.

Blood Vessels↗

Fibronectin, intercellular junctions and the sorting-out of chick embryonic tissue cells in monolayer.

A hierarchy of relative cohesiveness in monolayer of four different embryonic chick tissues was determined in a previous study. The hierarchy is: corneal epithelium congruent to liver parenchyma greater than pigmented epithelium greater than limb bud mesenchyme. The purpose of this paper is to describe the correlation between these adhesive relationships and, firstly, the amount of the adhesive glycoprotein, fibronectin, associated with the cells and, secondly, the morphology of their intercellular contacts. Fluorescent antibody staining of the cells with anti-fibronectin antibody showed that limb bud mesenchyme cells, the most weakly cohesive, had much more fibronectin than the other cell types. Thus there was a negative correlation between the amount of fibronectin and cellular cohesiveness. Analysis of intercellular contacts by electron microscopy showed that the most strongly cohesive cell types, corneal epithelium and liver parenchyma, were also those that possessed desmosomes.

Animals↗

The gastric mucosal barrier: structure of intercellular junctions in the dog.

The canine gastric mucosa consists of two regions, the surface mucous cells and gland area cells including parietal, chief, and mucous-containing cells. We have used quantitative freeze-fracture methods in conjunction with thin-section extracellular tracers to document and correlate tight junction morphology with epithelial permeability. The number of strands in the tight junction complexes of the surface cells and gland cells is the same, but differences in strand arrangement exist. The surface cells have an interwoven tight junction configuration which is impermeable to extracellular tracers. The gland cell junctions are regularly arranged and often permeable to extracellular lanthanum. The possibility that the observed difference in permeability between the tight junctions of the surface mucous cells and those of the gland cells is related to their structural configuration is discussed.

Animals↗

Intercellular junctions and tumor stage in small cell carcinoma of the lung.

The authors have studied the ultrastructural features of 52 cases of oat cell carcinoma of the lung and have related their observations to tumor stage and patient survival. Only the type of cell junctions seems to be of prognostic importance. Tumors with intermediate junctions--and especially those with desmosomes--have a more localized stage and may be resectable to result in longer survival than expected for oat cell carcinomas without junctions. For example, in the authors' series the median survival periods for those with no identifiable junctions, intermediate junctions, or desmosomes were 6.4, 8.2, and 11.3 months, respectively. Nevertheless, this ultrastructural subclassification is not as effective as that obtained from careful clinical staging.

Carcinoma, Small Cell↗

Biogenesis of structural intercellular junctions during cleavage in the mouse embryo.

The preimplantation embryo differentiates the trophectoderm epithelium which, from the 32-cell stage, generates the blastocoel of the blastocyst and, after implantation, gives rise to most extraembryonic lineages of the conceptus. Trophectoderm differentiation begins at compaction (8-cell stage) when cell-cell adhesion, mediated by uvomorulin, and epithelial cell polarisation first occur. Here, we review our work on the biogenesis of tight junctions and desmosomes during epithelial differentiation. Tight junction construction begins at compaction and appears to be a gradual process, both at morphological and molecular levels. This maturation pattern may be due in part to sequential expression of tight junction constituents from the embryonic genome. Tight junction formation is dependent upon uvomorulin adhesion but can be inhibited by different means without apparently disturbing cell adhesion or polarisation. Cell interactions appear to regulate tight junction tissue specificity, in part by controlling the level of synthesis of constituents. Desmosome formation begins at the 32-cell stage, particularly as the embryo initiates blastocoel accumulation, and, in contrast with tight junction formation, does not appear to be a gradual process. Thus, nascent desmosomes appear mature in terms of their molecular composition. Desmosomal proteins are synthesised well in advance of desmosome formation but the synthesis of the principal glycoprotein components begins at the blastocyst stage and may regulate the timing of junction assembly. Implications of these differing patterns of biogenesis for the embryo are discussed.

Animals↗

A fine structural analysis of intercellular junctions in the mouse liver.

Zonulae occludentes and gap junctions were examined both in the intact mouse liver and in a junction-rich membrane fraction from homogenized mouse liver. These preparations were visualized with the techniques of uranyl acetate staining en bloc, staining with colloidal lanthanum, negative staining with phosphotungstate, and freeze-cleaving. The zonula occludens is arranged as a meshwork of branching and anastomosing threadlike contacts sealing the lumen of the bile canaliculus from the liver intercellular space. The gap junction is characterized in section by a 20 A gap between the apposed junctional membrane outer leaflets, and permeation of this space with lanthanum or phosphotungstate reveals a polygonal lattice of subunits with a center-to-center spacing of 90-100 A. Freeze-cleaved gap junctions show a similar lattice. Extraction of junction-rich fractions with 60% aqueous acetone results in a disappearance of the 20 A gap in sectioned pellets and an inability to demonstrate the polygonal lattice with either the freeze-cleave or negative staining techniques. Extraction of the membranes with 50% acetone does not produce this effect. Thin-layer chromatography of the acetone extracts reveals a group of phospholipids in the 60% extract that are not detectable in the 50% extract. Acetone does not cause any detectable change in the structure of the zonula occludens, but the occluding junction becomes leaky to lanthanum following acetone treatment. The effects of other reagents on the junctions are reported.

Acetates↗

The induction of tumour cell adhesiveness and intercellular junctions by a glycoprotein of rat ascites hepatoma cell surface.

Rat ascites hepatoma AH109A cells (present as a free form in vivo) can aggregate and then develop well-defined tripartite junctional complexes, including intermediate junctions, desmosomes and focal tight junctions, on incubation with a glycoprotein separated from rat ascites hepatoma AH136B cells (forming cell islnds in vivo). The development of binding structures was strongly inhibited by actinomycin D. AH109A cells or rat ascites hepatoma YS cells (present as a free form in vivo) previously treated with the glycoprotein for 24 h, when inoculated i.p., proliferated as free cells in the ascitic fluid, like the untreated cells. AH109A cells actively proliferating in the skin do not form any junctional complexes. The reason for the failure of island formation by AH109A cells or YS cells in vivo is discussed.

Animals↗

Intercellular junctions in the organ of Corti as revealed by freeze fracturing.

The junctions between the cells of the organ of Corti have been investigated with the freeze-fracture method in the chinchilla inner ear. Numerous gap junctions have been found among all the supporting cells, particularly at the level of their basal part. This means that the supporting cells have their cytoplasm electrotonically and metabolically coupled. Gap junctions were not observed between supporting and sensory cells nor between sensory cells and nerve endings. At this level the transmission should be chemically mediated.

Animals↗

Intercellular junctions in the human developing preovulatory follicle and corpus luteum.

Using the lanthanum tracer and freeze-fracture techniques, the cell contacts of human ovarian cells were examined. The cell contacts between theca interna cells are similar to those seen between luteal cells are composed of septate-like cell contacts and gap junctions. On the other hand granulosa cells are connected by desmosomes and gap junctions. Thus, at least from the viewpoint of cell junctions, it is suggested that the main precursory cells of the luteal cells are the theca interna cells, although there is a general agreement that the luteal cells are derived from the granulosa cells.

Corpus Luteum↗

Exocrine pancreas under experimental conditions. IV. Alterations of intercellular junctions between acinar cells following pancreatic duct ligation.

Exocrine pancreatic tissue was investigated during various intervals after pancreatic duct ligation. Concomitant to the increase of the intraluminal pressure, alterations are found in the arrangement of the luminal membrane surface and the zonulae occludentes of the acinar cells. The zonulae occludentes exhibit a disarrangement of their strands. The number of strands diminishes and small interruptions as well as large discontinuities of the strands are observed in most regions of the acinar cells. Furthermore, gap junctions are found very infrequently between acinar cells. The ultrastructural alterations of the zonulae occludentes suggest a gradual adaptation to the increasing unilateral pressure. However, a leakage of the paracellular permeability barrier occurs which contributes to the known shunt between the compartments of the pancreatic juice and the interstitial space following pancreatic duct obstruction. The present investigation is a further example of alterations in the paracellular permeability and intercellular communication of epithelial cells under pathological conditions.

Animals↗

Intercellular junctions and the development of the blood-brain barrier in Manduca sexta.

In early embryonic development of the tobacco horn moth no blood-brain barrier is present, as shown by the unimpeded entry of exogenous tracers into the nervous system. However, later on, just before hatching, lanthanum and horseradish peroxidase (HRP) are unable to move inwardly beyond the level of the perineurium, which is the morphological site of the blood--brain barrier in the adult moth, as well as in other insects. Freeze-fracture studies indicate that in the early embryo, 10 nm particles are scattered about in the perineurial membrane PF, either as separate entities or as short linear arrays. By hatching or just before, however, the 10 nm particles have become aligned into lengthy linear aggregates as PF ridges with EF grooves. These would appear to be the simple, arthropod-form of tight junction, and are presumed to be the basis of the perineurial blood-brain barrier. At about the same time, gap junctional elements appear both between adjacent perineurial cells and between glial cells. In both cell types, the gap junctions form from free 13 nm EF particles which gradually become aligned or clumped into strands and aggregates which ultimately coalesce to form first irregular masses and then the macular plaques typical of mature gap junctions. Many of the latter stages are coincident with the hatching of a motile larvae, so that the perineurial and glial cells are by this stage coupled via the channels of the gap junctional particles. They are therefore able to undergo both ionic and metabolic exchange and cooperation during larval life, in addition to being able to respond to hormonal substances in an integrated way. During the 5 larval instars more gap junctions form as the perineurial layer grows thicker. These junctions become more regular in outline and their particles more tightly packed; these larval structures are compared with junctions found in the adult which tend to be more extensive but otherwise similar. Since no septate junctions are apparent during Manduca embryonic or larval life when the blood-brain barrier forms, nor in adults, the results of this report support the contention that it is the tight junctions rather than septate ones which form the basis of permeability barriers in this, and probably other, arthropod systems.

Animals↗