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PubMed1975

Effect of temporary hypoxia on the permeability of the rat aorta.

Hypoxia was produced experimentally in the infrarenal portion of rat abdominal aorta by ligating it at both ends for one hour. Permeability changes of the injured segment were followed up by means of a colloidal iron tracer after different periods of recirculation. The tracer could be demonstrated in intercellular junctions and subendothelial space already after one hour of recirculation. The permeability disorder was at its peak two days after the injury and it ceased by the tenth day. Insudation of the media by plasma increased proportionally with the endothelial permeability change and it was accompanied by marked ultrastructural alterations.

Animals↗
PubMed1979

Relation of capillary morphology to transport of fluid and large molecules: a review.

Transport characteristics of endothelial cell membranes, cytoplasmic vesicles, intercellular junctions (small pores and large pores) and fenestrae are presented in terms of appropriate models. Morphological differences of capillaries in different vascular beds are compared with physiological differences in permeability to water, small molecules and large molecules, and the correlation of morphologically defined and physiologically defined pathways is analyzed.

Animals↗
PubMed1996

"Cytosolic" phospholipase A2 is in the nucleus of subconfluent endothelial cells but confined to the cytoplasm of confluent endothelial cells and redistributes to the nuclear envelope and cell junctions upon histamine stimulation.

The synthesis of arachidonic acid metabolites is initiated by activation of the sn-arachidonyl-dependent, 85-kd "cytosolic" phospholipase A2 (cPLA2) enzyme. We have investigated the subcellular localization of cPLA2 in resting and histamine-treated human and bovine endothelial cells (EC) using confocal immunofluorescence microscopy. In tightly confluent EC, cPLA2 was primarily localized in the cytoplasm. Surprisingly, in subconfluent EC, cPLA2 was also prominently located within the cell nucleus. By immunoblotting of cell lysates after SDS-PAGE, the cytoplasmic molecular species in subconfluent cells displayed the characteristic Mr 110,000, whereas nuclear extracts contained a predominant Mr 70,000. Nuclear localization of cPLA, in subconfluent EC is independent of cell cycle because it was observed in growth-arrested cells as well as in dividing cells. Nuclear localization was also observed in subconfluent MDCK and HeLa cells where, in contrast to EC, it persisted in tightly confluent monolayers. Treatment of subconfluent EC with histamine caused a rapid, dose-dependent redistribution of cPLA2, from the nucleus to the nuclear envelope. The same treatment of confluent EC produced translocation of cytoplasmic enzyme to the nuclear envelope and to the plasma membrane at the intercellular junctions. The cell density dependence of cPLA2, localization may contribute to altered arachidonic acid metabolism in injured vessels as compared with quiescent vessels.

Animals↗
PubMed1982

Reactivation of intestinal epithelial cell brush border motility: ATP-dependent contraction via a terminal web contractile ring.

Various models have been put forward suggesting ways in which brush borders from intestinal epithelial cells may be motile. Experiments documenting putative brush border motility have been performed on isolated brush borders and have generated models suggesting microvillar retraction or microvillar rootlet interactions. The reported Ca++ ATP-induced retraction of microvilli has been shown, instead, to be microvillar dissolution in response to Ca++ and not active brush border motility. I report here studies on the reactivation of motility in intact sheets of isolated intestinal epithelium. Whole epithelial sheets were glycerinated, which leaves the brush border and intercellular junctions intact, and then treated with ATP, PPi, ITP, ADP, GTP, or delta S-ATP. Analysis by video enhanced differential interference-contrast microscopy and thin-section transmission electron microscopy reveals contractions in the terminal web region causing microvilli to be fanned apart in response to ATP and delta S-ATP but not in response to ADP, PPi, ITP, or GTP. Electron microscopy reveals that the contractions occur at the level of the intermediate junction in a circumferential constriction which can pull cells completely apart. This constriction occurs in a location occupied by an actin-containing circumferential band of filaments, as demonstrated by S-1 binding, which completely encircles the terminal web at the level of the intermediate junction. Upon contraction, this band becomes denser and thicker. Since myosin, alpha-actinin and tropomyosin, in addition to actin, have been localized to this region of the terminal web, it is proposed that the intestinal epithelial cell can be motile via a circumferential terminal web contractile ring analogous to the contractile ring of dividing cells.

Adenosine Triphosphate↗
PubMed1975

Studies on the juxtaglomerular apparatus. IV. Freeze-fracturing of membrane surfaces.

The juxtaglomerular apparatus of the rat was studied after freeze-fracturing with special respect to intercellular junctions. It was found that juxtaglomerular granulated cells of the vas afferens are interconnected by gap junctions to adjacent cells (granulated cells, possibly also smooth muscle cells). Gap junctions have also been found on the surface of lacis cells and mesangial cells. It is therefore concluded that these cells of the juxtaglomerular apparatus and the glomerulus--granulated cells (possibly also smooth muscle cells) of the vas afferens, lacis cells and mesangium cells--form a functional system reacting in a coordinated manner to physiological stimuli.

Animals↗
PubMed1979

An ultrastructural study of cultured human meningioma cells.

Ultrastructural alterations in human meningioma cells growth in vitro are reported. In early passages the cells retain some of the characteristic features of the original tumors. These include interdigitations associated with intercellular junctional devices (e.g., desmosomes, gap junctions). However, with repeated subculture these features tend to be less frequent. Typical whorl formations are observed only in primary cultures. The number of cytoplasmic filaments, lipid inclusions and other dense bodies increases with time in culture. Cytoplasmic invaginations into nuclei and the appearance of very large cells become more frequent in repeatedly transferred cultures.

Cells, Cultured↗
PubMed1975

Vascular structures in brain tumors.

The blood vessels within brain tumors show alterations from the normal anatomy. Some of these seem to be related to an increased capacity to transfer materials between the lumen and the parenchyma and are probably intimately connected with the edema associated with the tumor. These alterations include fenestration, widened intercellular junctions, increase in pinocytotic vesicles, and infolding of the luminal surface. Other alterations are observed but their function is not as clear. The latter include an increase in the number of tubular bodies, the appearance of tubular structures within vacuoles, tubular arrays within the nuclear envelope and rough endoplasmic reticulum, and endothelial proliferation, among others.

Basement Membrane↗
PubMedDate not supplied

Olfactory neuroepithelioma arising from the olfactory placode.

The patient was a 54-year-old man, who had lost his sense of smell 6 years previously and had started to become forgetful about 6 months prior to presenting at hospital. MRI admission showed a large multicystic tumor with Gd-DTPA enhancement extending from the anterior cranial fossa through the sphenoid sinus and into the nasal cavity. Histopathological examination revealed extensive proliferation of small round cells that were divided by connective tissue septae. The tumor cells occasionally formed tubular structures, although no basement membranes were present. On immunostaining, round tumor cells were positive for neuron-specific enolase, synaptophysin, and chromogranin A, while cells forming tubules were positive for AE 1 and CAM 5.2. Almost all of the tumor cells were positive for Ber-EP4, and some of the epithelioid cells surrounding the tubular structures were also positive for luteinizing hormone-releasing hormone (LH-RH). Electron microscopy demonstrated sporadic intercellular junctions, many microtubules in the tumor cell processes, and clear- and dense-cored vesicles in the cytoplasm. Based on the results, this case appears to be the first documented neuroepithelioma with Ber-EP4- and LH-RH-positive cells arising from the olfactory placode.

Biomarkers, Tumor↗
PubMed1990

Cell-cell interactions in the process of differentiation of thyroid epithelial cells into follicles: a study by microinjection and fluorescence microscopy on in vitro reconstituted thyroid follicles.

Thyroid cells, cultured in the presence of thyroid stimulating hormone, reorganized within 36-48 hr into follicular structures, the in vitro reconstituted thyroid follicles or RTF. By microinjection of fluorescent probes either into the neoformed intrafollicular lumen (IL) or into cells forming the follicles, we have studied the development and some functional properties of cell-cell contacts involved in a) the formation of the thyroid follicular lumen and b) the communication between thyrocytes within the follicle. The probes were compounds of either low (Lucifer Yellow: LY) or high molecular weight (Dextran labeled with fluorescein: FITC-Dextran and Cascade Blue conjugated to bovine serum albumin: CB-BSA). LY microinjected into IL of 2-9-day-old RTF was seen to label circular spaces with a diameter ranging from 10 to 100 microns. The cells delimiting the IL remained unlabeled. The fluorescent dye remained concentrated in IL for up to 24 hr. FITC-Dextran or CB-BSA microinjected into IL behaved as LY; the probes were restrained into the lumen. A 2 hr incubation of RTF with iodide induced alterations of the structure of IL; an effect mediated by an organic form of actively trapped iodide. A 15-30 min incubation of RTF in a low CA2+ medium caused the opening of IL visualized by the progressive decrease of the fluorescence of probes preinjected into the lumenal space. The same but more rapid effect was obtained by microinjection of EGTA into the IL. The low Ca2(+)-dependent opening of IL was also demonstrated by the release into the medium of thyroglobulin present in IL. Microinjection of LY in a cell involved in the follicle structure led to the rapid labeling of the other cells forming the follicle but LY did not penetrate the IL. Unlike LY, the distribution of FITC-Dextran or CB-BSA injected into cells delimiting the lumen was restricted to the microinjected cells. Alterations of medium or intralumenal Ca2+ concentration which caused the opening of IL did not affect the cell-to-cell transfer of LY. By using fluorescent probe microinjection, we show that the in vitro thyrocyte histiotypic differentiation leads to the reconstitution of functional intercellular junctions: tight junctions insuring the tightness of the neoformed lumen and gap junctions mediating the cell-to-cell exchange of small molecules. The structure of the thyroid follicles appears to be under the control of both extracellular and intralumenal Ca2+ concentrations.

Acridine Orange↗
PubMed1981

Cellular interrelationships in the human fetal ovary and testis.

Information on the cellular changes involved in human gonadal maturation has increased considerably in recent years with the introduction of special techniques for coordinating structural and functional analyses. An approach that appears to be particularly promising is the use of freeze-fracture techniques. Utilizing this method, we have demonstrated the formation of specialized intercellular junctions in the developing ovary and testis. Studies in several species have indicated similar findings which relate to the formation of the blood-testis barrier and the coordination of cellular activity involved in steroidogenesis. Further progress in understanding of maturational changes in the developing gonads will depend on continuing correlation of structural and functional changes in specific cell populations.

Cell Differentiation↗
PubMedDate not supplied

Transepithelial migration of neutrophils.

Neutrophils are a key cell type of nonadaptive immune system and are the first phagocytic cell type that reaches mucosal inflammatory sites. On the last stage of their journey from the blood stream to a mucosal surface, neutrophils cross a generally sealed epithelium by migrating along the paracellular pathway to the luminal side of the epithelial layer. This last step involves a specific receptor-mediated adhesion event of the neutrophil to the epithelium, followed by a rapid and highly coordinated reversible opening of the epithelial intercellular junctions that allows the transmigration of the neutrophils. Although we do not yet understand the molecular mechanisms that mediate this transmigration process, the last years witnessed the discovery of the first neutrophil and epithelial cell surface proteins critically involved in transepithelial migration of neutrophils.

Animals↗
PubMed1995

Alterations in cell-cell communication in human papillomavirus type 16 (HPV16) transformed rat myoblasts.

A reduction of gap-junctional intercellular communication (GJIC) often accompanies neoplastic transformation. The present work demonstrates that transformation by the oncogenic human DNA virus, human papilloma virus 16(HPV16), also reduces GJIC between L6 rat myoblasts. HPVs are associated with anogenital cancers, the incidence of which is increasing in HIV positive patients of both sexes. Using videofluorescence imaging of Fura-2 loaded cells a lack of GJIC between transformed HPV16-L6 cells was first indicated by uncoordinated brief [Ca2+]i spikes in clusters of DMSO-treated HPV16-L6 cells instead of the synchronous, sustained [Ca2+]i surges in clusters of DMSO-treated L6 cells. Reduced GJIC between HPV16-L6 cells was demonstrated directly by a much reduced transfer of lucifer yellow dye from HPV16-L6 cells, which had been loaded with the dye through electroporation with an EPIZAP II in situ electroporator, to neighbouring nonelectroporated HPV16-L6 cells. One reason for this reduced GJIC between HPV16-L6 cells could have been their dramatically enhanced activity of membrane-associated PKC which is known to phosphorylate connexins and down-regulate gap junctions. However, the main reason was the viral-induced inhibition of the expression of a major gap junction component, Cx43 (Connexin 43), in the transformed myoblasts.

Animals↗
PubMed1984

Comparative studies on the ultrastructure of malignant melanoma in fish and human by freeze-etching and transmission electron microscopy.

Malignant melanomas (MM) in the fish Xiphophorus and in humans were studied both by transmission electron microscopy (TEM) and freeze-etching (FE). In both fish and human melanomas the cells show interdigitations of the plasma membranes. The nuclei are large and lobulated and have many nuclear pores. Melanosomes are abundant and melanosome complexes ("compound melanosomes") occur regularly. Pinocytotic vesicles could be demonstrated in fish and human melanomas showing local differences in frequency and distribution patterns in the tumor. Intercellular junctions are lacking in MM cells from fish and humans. The FE technique showed considerable advantages in demonstrating membrane-surface peculiarities such as nuclear pores or pinocytotic vesicles. The FE replicas of fish melanomas are like those of humans. These findings may support the hypothesis that melanoma in fish and humans reflect the same biological phenomenon.

Animals↗
PubMed2003

Mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells.

VE-cadherin is an endothelial-specific cadherin that plays important roles in vascular morphogenesis and growth control. To investigate the mechanisms by which endothelial cells regulate cadherin cell surface levels, a VE-cadherin mutant containing the non-adhesive interleukin-2 (IL-2) receptor extracellular domain and the VE-cadherin cytoplasmic tail (IL-2R-VE-cadcyto) was expressed in microvascular endothelial cells. Expression of the IL-2R-VE-cadcyto mutant resulted in the internalization of endogenous VE-cadherin and in a dramatic decrease in endogenous VE-cadherin levels. The internalized VE-cadherin co-localized with early endosomes, and the lysosomal inhibitor chloroquine dramatically inhibited the down-regulation of VE-cadherin in cells expressing the IL-2R-VE-cadcyto mutant. Chloroquine treatment also resulted in the accumulation of a VE-cadherin fragment lacking the beta-catenin binding domain of the VE-cadherin cytoplasmic tail. The formation of the VE-cadherin fragment could be prevented by treating endothelial cells with proteasome inhibitors. Furthermore, inhibition of the proteasome prevented VE-cadherin internalization and inhibited the disruption of endothelial intercellular junctions by the IL-2RVE-cadcyto mutant. These results provide new insights into the mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells.

Adenoviridae↗
PubMed1983

Ultrastructural effects of cadmium on the rat epididymis.

The short-term effect of a subcutaneous injection of 3 mumol of cadmium chloride/100 g body weight were examined 90 min later in male rats. The main target of cadmium, as shown by electron microscopy, was the endothelial lining of capillaries of the caput epididymidis. Intercellular junctions between endothelial cells were disorganized, ranging from slight separation between cytoplasmic leaflets to wide gaps which communicated freely with the pericapillary tissue. Disruption of platelets and intravascular clotting followed these early endothelial lesions, and clotting was found in venules as an extension of clotting from the capillaries. Pre-treatment with zinc (300 mumol zinc chloride/100 g body weight) prevented all of the lesions described above.

Animals↗
PubMed1996

The intercalated disc of monkey myocardial cells and Purkinje fibers as revealed by scanning electron microscopy.

The intercalated discs of working myocardium and Purkinje fibers of the monkey heart were examined by scanning and transmission electron microscopy. The NaOH/ultrasonication technique resulted in the digestion of connective tissue and a separation of the intercellular junctions of intercalated discs, such that these could be visualized three-dimensionally. The intercalated discs of ventricular myocytes, atrial myocytes and Purkinje fibers vary considerably in number and configuration, as do the intercalated discs of the three different layers of the ventricular myocardium. Myocytes in the subepicardial, middle and subendocardial layers of the ventricle have 1-3, 4-5 and 5-6 intercalated discs at the end of these cells, respectively. Those in the endocardial layer are characterized by the presence of small laterally-placed intercalated discs. Atrial myocytes and Purkinje fibers usually only have 1-2 intercalated discs. Individual intercalated discs in ventricular myocytes have complicated stairs with 10-30 steps and corresponding risers, while those of atrial myocytes and Purkinje fibers have simple stairs with 1-3 steps and risers. Steps equivalent to the plicate segments are characterized by densely-packed microplicae and finger-like microprojections which greatly increase surface area in ventricular myocytes. Microprojections in atrial myocytes and Purkinje fibers are sparse by comparison. Risers equivalent to the interplicate segments containing large gap junctional areas are most numerous in left ventricular myocytes, followed by right ventricular myocytes, Purkinje fibers and atrial myocytes in decreasing order. The geometric arrangement of the various types of myocytes may be related with impulse propagation. Large intercalated discs of cell trunks and series branches may participate in longitudinal propagation, while small laterally-placed ones may be the site of transverse propagation.

Animals↗
PubMed1984

Ultrastructural aspects of the histogenesis of diffuse and localized mesothelioma.

During an ultrastructural review of 30 diffuse and 10 localized mesotheliomas, it was apparent that some micrographs showed various stages in the developmental processes involved in the formation of histological patterns in diffuse mesotheliomas and a histogenetic link between diffuse and localized mesotheliomas. Cells in the stromal or sarcomatous regions of diffuse mesothelioma often show varying degrees of mesothelial differentiation and a gradual transition to cells with typical mesothelial characteristics that organize into structures recapitulating the surface layer of serosal membranes. Tumor cells in localized mesotheliomas had many similarities to the "stromal" cells in the diffuse counterpart including intercellular junctions, rare microvilli and occasional foci of basal lamina. It is postulated that diffuse and localized mesotheliomas share a common histogenetic origin as a result of neoplastic induction of specialized submesothelial cells. In this concept, tumor cells in diffuse mesotheliomas reflect stages in the differentiation and organization of normal serosal membranes and localized mesotheliomas mirror the earliest phases of this process.

Humans↗
PubMed1994

Tumor-suppressor gene products in cell contacts: the cadherin-APC-armadillo connection.

Various structural components of intercellular junctions have recently been found to represent (or be related to) products of tumor-suppressor genes. The tumor-suppressor gene product adenomatous polyposis coli (APC) binds to beta 2-catenin (homologous to the product of Drosophila armadillo), which is cytoplasmically associated with the cell adhesion molecule E-cadherin.

Adenomatous Polyposis Coli Protein↗