scholarly journals Intercellular junctions and transfer of small molecules in primary vascular endothelial cultures.

1982 ◽  
Vol 92 (1) ◽  
pp. 183-191 ◽  
Author(s):  
D M Larson ◽  
J D Sheridan
Keyword(s):  
Gap Junctions ◽  
Small Molecules ◽  
Tight Junctions ◽  
Lucifer Yellow ◽  
Primary Cultures ◽  
Freeze Fracture ◽  
Cell Transfer ◽  
Isolated Cells ◽  
Thin Sections ◽  
Cell To Cell Transfer

The ultrastructure of gap and tight junctions and the cell-to-cell transfer of small molecules were studied in primary cultures and freshly isolated sheets of endothelial cells from calf aortae and umbilical veins. In thin sections and in freeze-fracture replicas, the gap and tight junctions in the freshly isolated cells from both sources appeared similar to those found in the intimal endothelium. Most of the interfaces in replicas had complex arrays of multiple gap junctions either intercalated within tight junction networks or interconnected by linear particle strands. The particle density in the center of most gap junctions was noticeably reduced. In confluent monolayers, after 3-5 days in culture, gap and tight junctions were present, although reduced in complexity and apparent extent. Despite the relative simplicity of the junctions, the cell-to-cell transfer of potential changes, dye (Lucifer Yellow CH), and nucleotides was readily detectable in cultures of both endothelial cell types. The extent and rapidity of dye transfer in culture was only slightly less than that in sheets of freshly isolated cells, perhaps reflecting a reduced gap junctional area combined with an increase in cell size in vitro.

Gap junctions in the prothoracic glands of the tobacco hornworm, Manduca sexta

1992 ◽  
Vol 50 (1) ◽  
pp. 706-707
Author(s):  
Ji-da Dai ◽  
M. Joseph Costello ◽  
Lawrence I. Gilbert
Keyword(s):  
Gap Junctions ◽  
Small Molecules ◽  
Manduca Sexta ◽  
Freeze Fracture ◽  
Cell Communication ◽  
Cellular Level ◽  
Thin Sections ◽  
Prothoracic Glands ◽  
Polyhydroxylated Steroids ◽  
Stimulation Of

Insect molting and metamorphosis are elicited by a class of polyhydroxylated steroids, ecdysteroids, that originate in the prothoracic glands (PGs). Prothoracicotropic hormone stimulation of steroidogenesis by the PGs at the cellular level involves both calcium and cAMP. Cell-to-cell communication mediated by gap junctions may play a key role in regulating signal transduction by controlling the transmission of small molecules and ions between adjacent cells. This is the first report of gap junctions in the PGs, the evidence obtained by means of SEM, thin sections and freeze-fracture replicas.

Contacts between pigmented retina epithelial cells in culture

1976 ◽  
Vol 22 (2) ◽  
pp. 371-383 ◽  
Author(s):  
C.A. Middleton ◽  
S.M. Pegrum
Keyword(s):  
Gap Junctions ◽  
Tight Junctions ◽  
Primary Cultures ◽  
Embryonic Chick ◽  
Isolated Cells ◽  
Gradual Reduction ◽  
Extensive Region ◽  
Cytoplasmic Filaments ◽  
Junctional Complexes ◽  
Number Of Cells

The behaviour of primary cultures of dissociated embryonic chick pigmented retina epithelial (PRE) cells has been investigated. Isolated PRE cells have a mean speed of locomotion of 7–16 mum/h. Collisions between the cells normally result in the development of stable contacts between the cells involved. This leads to a gradual reduction in the number of isolated cells and an increase in the number of cells incorporated into islands. Ultrastructural observations of islands of cells after 24 h in culture show that junctional complexes are present between the cells. These complexes consist of 2 components: (a) an apically situated region of focal tight junctions and/or gap junctions, and (b) a more ventrally located zonula adhaerens with associated cytoplasmic filaments forming a band running completely around the periphery of each cell. The intermembrane gap in the region of the zonula is 6-0-12-0 nm. The junctional complexes become more differentiated with time and after 48 h in culture consist of an extensive region of tight junctions and/or gap junctions and a more specialized zonula adhaerens. It is suggested that the development of junctional complexes may be responsible for the stable contacts that the cells display in culture.

scholarly journals Cell junctions and locomotion of the blastoderm edge in gastrulating chick and quail embryos

1985 ◽  
Vol 78 (1) ◽  
pp. 191-204
Author(s):  
L. Andries ◽  
F. Harrisson ◽  
R. Hertsens ◽  
L. Vakaet
Keyword(s):  
Gap Junctions ◽  
Tight Junctions ◽  
Functional Organization ◽  
Freeze Fracture ◽  
Leading Edge ◽  
Proximal Part ◽  
Cell Junctions ◽  
Vitelline Membrane ◽  
Thin Sections ◽  
Marginal Cells

The blastoderm edge migrates by the active locomotion of a multilayer of epithelial cells, the so-called margin of overgrowth (MO), that uses the vitelline membrane as its substratum. The structural unity formed by the margin of overgrowth cells and their rapid migration suggest coordination of locomotion between individual cells. Using transmission electron microscopy of thin sections and freeze-fracture, we attempted to determine if the pattern of junctions of the migrating margin of overgrowth is related to the suggested cell—cell cooperation between individual cells in this region. In the leading edge there are large areas of closely apposed cell membranes. Incipient desmosomes and small gap junctions were observed. Tight junctions consisted of isolated strands or isolated networks of tight-junctional strands. In the proximal part of the margin of overgrowth the size of the gap junctions increased and the desmosomes were fully developed. Tight-junctional strands were either isolated or arranged into an isolated network. A broad belt of tight junctions was observed at the transition between margin of overgrowth and non-marginal cells. The distribution of the junctional elements in the MO suggests that junctions contribute to the maintenance of the structural and functional organization of the margin of overgrowth. Furthermore, the spatial distribution of the junctions might give information about the mechanism of locomotion of the margin of overgrowth.

The application of freeze-fracture in the analysis of intercellular junctions in pleuro-pulmonary tumors

1990 ◽  
Vol 48 (3) ◽  
pp. 540-541
Author(s):  
T. M. Mukherjee ◽  
J. G. Swift
Keyword(s):  
Gap Junctions ◽  
Tight Junctions ◽  
Thin Section ◽  
Freeze Fracture ◽  
Intercellular Junctions ◽  
Squamous Cell Carcinomas ◽  
Cell Junctions ◽  
Thin Sections ◽  
Pulmonary Tumors ◽  
Diagnostic Importance

Thin section and freeze-fracture techniques have been used to examine the morphology of cell junctions in a variety of pleuro-pulmonary tumours with the aim of identifying features that may be of diagnostic importance or of significance in the development of the tumour. Freeze-fracture preparations are particularly useful for the analysis of cell junctions, since extensive face views of the interior of the cell membrane are exposed. This enables precise characterisation of the type of junctions present, their extent and their inter-relationships.Freeze-fracture replicas can reveal the presence of junctions that would be difficult or impossible to detect in thin sections. For example, desmosomes are a well-known feature in thin sections of squamous cell carcinomas, but these tumours may also have focal tight junctions and gap junctions (Figs. 1,2). The tight and gap junctions can occur separately (Fig.l), or in combination (Fig. 2). Similarly, in a recent study of a case of “Ewing’s sarcoma”, replicas showed the presence of unusual, elaborate focal tight junctions, a feature never suspected from the routine thin section studies of this tumour.

scholarly journals Gap junction structures after experimental alteration of junctional channel conductance.

1985 ◽  
Vol 101 (5) ◽  
pp. 1741-1748 ◽  
Author(s):  
T M Miller ◽  
D A Goodenough
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Electron Micrographs ◽  
Time Course ◽  
Structural Changes ◽  
Lucifer Yellow ◽  
Freeze Fracture ◽  
Channel Conductance ◽  
Experimental Alteration ◽  
Quick Freezing

Gap junctions are known to present a variety of different morphologies in electron micrographs and x-ray diffraction patterns. This variation in structure is not only seen between gap junctions in different tissues and organisms, but also within a given tissue. In an attempt to understand the physiological meaning of some aspects of this variability, gap junction structure was studied following experimental manipulation of junctional channel conductance. Both physiological and morphological experiments were performed on gap junctions joining stage 20-23 chick embryo lens epithelial cells. Channel conductance was experimentally altered by using five different experimental manipulations, and assayed for conductance changes by observing the intercellular diffusion of Lucifer Yellow CH. All structural measurements were made on electron micrographs of freeze-fracture replicas after quick-freezing of specimens from the living state; for comparison, aldehyde-fixed specimens were measured as well. Analysis of the data generated as a result of this study revealed no common statistically significant changes in the intrajunctional packing of connexons in the membrane plane as a result of experimental alteration of junctional channel conductance, although some of the experimental manipulations used to alter junctional conductance did produce significant structural changes. Aldehyde fixation caused a dramatic condensation of connexon packing, a result not observed with any of the five experimental uncoupling conditions over the 40-min time course of the experiments.

scholarly journals VARIATIONS IN TIGHT AND GAP JUNCTIONS IN MAMMALIAN TISSUES

1972 ◽  
Vol 53 (3) ◽  
pp. 758-776 ◽  
Author(s):  
Daniel S. Friend ◽  
Norton B. Gilula
Keyword(s):  
Smooth Muscle ◽  
Gap Junctions ◽  
Tight Junctions ◽  
Adrenal Cortex ◽  
Freeze Fracture ◽  
Junctional Complex ◽  
Exocrine Glands ◽  
Rat Pancreas ◽  
Zonula Occludens ◽  
Mammalian Tissues

The fine structure and distribution of tight (zonula occludens) and gap junctions in epithelia of the rat pancreas, liver, adrenal cortex, epididymis, and duodenum, and in smooth muscle were examined in paraformaldehyde-glutaraldehyde-fixed, tracer-permeated (K-pyroantimonate and lanthanum), and freeze-fractured tissue preparations. While many pentalaminar and septilaminar foci seen in thin-section and tracer preparations can be recognized as corresponding to well-characterized freeze-fracture images of tight and gap junction membrane modifications, many others cannot be unequivocally categorized—nor can all freeze-etched aggregates of membrane particles. Generally, epithelia of exocrine glands (pancreas and liver) have moderate-sized tight junctions and large gap junctions, with many of their gap junctions basal to the junctional complex. In contrast, the adrenal cortex, a ductless gland, may not have a tight junction but does possess large gap junctions. Mucosal epithelia (epididymis and intestine) have extensive tight junctions, but their gap junctions are not as well developed as those of glandular tissue. Smooth muscle contains numerous small gap junctions The incidence, size, and configuration of the junctions we observed correlate well with the known functions of the junctions and of the tissues where they are found.

scholarly journals Connexin-specific cell-to-cell transfer of short interfering RNA by gap junctions

2005 ◽  
Vol 568 (2) ◽  
pp. 459-468 ◽  
Author(s):  
V. Valiunas ◽  
Y. Y. Polosina ◽  
H. Miller ◽  
I. A. Potapova ◽  
L. Valiuniene ◽  
...  
Keyword(s):  
Gap Junctions ◽  
Specific Cell ◽  
Cell Transfer ◽  
Short Interfering Rna ◽  
Interfering Rna ◽  
Cell To Cell Transfer

scholarly journals A freeze-fracture study on the developing satellite cells of spinal ganglia in the chick embryo

1988 ◽  
Vol 46 (1) ◽  
pp. 6-9
Author(s):  
Claudio A. Ferraz de Carvalho ◽  
Ciro F. da Silva
Keyword(s):  
Chick Embryo ◽  
Gap Junctions ◽  
Tight Junctions ◽  
Small Groups ◽  
Satellite Cells ◽  
Freeze Fracture ◽  
Fracture Analysis ◽  
Spinal Ganglia ◽  
Pinocytotic Vesicles ◽  
Fracture Study

A freeze-fracture analysis of the satellite cells of spinal ganglia of the chick embryo was performed in 8 successive stages of development, from the 5th incubation day to hatching. The characteristic laminar disposition of the cells were first observed on the 7th day. Tight junctions were found at the 20th incubation day. Small groups or irregular aggregates of particles, but not gap junctions, were described on the 7th and 8th days. Pinocytotic vesicles were pointed out in the different stages considered.

Morphological Changes of Intercellular Junctions in the Rat Submandibular Gland Treated by Long-term Repeated Administration of Isoproterenol

1987 ◽  
Vol 66 (8) ◽  
pp. 1303-1309 ◽  
Author(s):  
T. Inoue ◽  
H. Yamane ◽  
T. Yamamura ◽  
M. Shimono
Keyword(s):  
Gap Junctions ◽  
Tight Junctions ◽  
Morphological Changes ◽  
Freeze Fracture ◽  
Acinar Cells ◽  
Intercellular Junctions ◽  
Repeated Administration ◽  
Experimental Conditions ◽  
Significant Difference

Long-term repeated administration of isoproterenol (lPR) 2 mg/100 g bw, once daily for ten days, resulted in morphological changes in the intercellular junctions of rat submandibular glands, which were investigated by means of the freeze fracture technique. A significantly increased number of tight-junctional strands was present. These junctional strands extended much deeper toward the basal membrane than those in normal acinar cells. The basal frontier strands that branched from the networks of tight junctions were elongated and had either free-endings or terminal loops, which were more frequently observed in the IPR-treated acinar cells than in untreated acinar cells. Some of the strands of tight junctions were connected to small gap junctions. The diameters of gap junctions were not significantly different from those of control acinar cells. However, smooth areas devoid of particles were found intermingling with the usual packed particles in irregularly shaped small gap junctions. There was no significant difference between the desmosomes of IPR-treated and untreated acinar cells, in terms of either morphology or distribution. These changes in junctional morphology in the IPR-treated acinar cells resemble those seen in salivary glands during development, and in some experimental conditions including tumorous changes.

scholarly journals Occluding junctions and cell behavior in primary cultures of normal and neoplastic mammary gland cells.

1975 ◽  
Vol 66 (2) ◽  
pp. 316-332 ◽  
Author(s):  
P B Pickett ◽  
D R Pitelka ◽  
S T Hamamoto ◽  
D S Misfeldt
Keyword(s):  
Primary Cultures ◽  
Freeze Fracture ◽  
Culture Conditions ◽  
Pavement Cells ◽  
Thin Sections ◽  
Occluding Junctions ◽  
Bulk Medium ◽  
Mammary Gland Cells ◽  
Junction Structure

Cells dissociated from normal prelactating mouse mammary glands or from spontaneous mammary adenocarcinomas, when grown at high density on an impermeable substrate, form nonproliferating, confluent, epithelial pavements in which turgid, blister-like domes appear as a result of fluid accumulation beneath the cell layer. To compare the structure of the fluid-segregating cell associations in normal and tumor cell cultures with that of lactating gland in vivo, we have examined such cultures alive and in thick and thin sections and freeze-fracture replicas. Pavement cells in all cases are polarized toward the bulk medium as a lumen equivalent, with microvilli and continuous, well-developed occluding junctions at this surface. Between the pavement and the substrate are other cells, of parenchymal or stromal origin, scattered or in loose piles; these sequestered cells are relatively unpolarized and never possess occluding junctions. Small gap junctions have been found in the pavement layer, and desmosomes may link epithelial cells in any location. Under the culture conditions used, development of the epithelial secretory apparatus is not demonstrable; normal and neoplastic cells do not differ consistently in any property examined. A dome's roof is merely a raised part of the epithelial pavement and does not differ from the latter in either cell or junction structure. We suggest that dome formation demonstrates the persistence of some transport functions and of the capacity to form effective occluding junctions. These basic epithelial properties can survive both neoplastic transformation and transition to culture.

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