scholarly journals A freeze-fracture and lanthanum tracer study of the complex junction between Sertoli cells of the canine testis.

1978 ◽  
Vol 76 (1) ◽  
pp. 57-75 ◽  
Author(s):  
C J Connell
Keyword(s):  
Tight Junctions ◽  
Sertoli Cells ◽  
Freeze Fracture ◽  
Intercellular Junctions ◽  
Septate Junction ◽  
Septate Junctions ◽  
Thin Sections ◽  
En Face ◽  
Lanthanum Tracer ◽  
Complex Junction

What appear to be true septate junctions by all techniques currently available for the cytological identification of intercellular junctions are part of a complex junction that interconnects the Sertoli cells of the canine testis. In the seminiferous epithelium, septate junctions are located basal to belts of tight junctions. In thin sections, septate junctions appear as double, parallel, transverse connections or septa spanning an approximately 90-A intercellular space between adjacent Sertoli cells. In en face sections of lanthanum-aldehyde-perfused specimens, the septa themselves exclude lanthanum and appear as electron-lucent lines arranged in a series of double, parallel rows on a background of electron-dense lanthanum. In freeze-fracture replicas this vertebrate septate junction appears as double, parallel rows of individual or fused particles which conform to the distribution of the intercellular septa. Septate junctions can be clearly distinguished from tight junctions as tight junctions prevent the movement of lanthanum tracer toward the lumen, appear as single rows of individual or fused particles in interlacing patterns within freeze-fracture replicas, and are seen as areas of close membrane apposition in thin sections. Both the septate junction and the tight junction are associated with specializations of the Sertoli cell cytoplasm. This is the first demonstration in a vertebrate tissue of a true septate junction.

Evidence for septate junctions in the syzygy of the protozoon Gregarina polymorpha (Protozoa, Apicomplexa)

1988 ◽  
Vol 89 (2) ◽  
pp. 217-224
Author(s):  
ROMANO DALLAI ◽  
MARIA VEGNI TALLURI
Keyword(s):  
Plasma Membrane ◽  
Intercellular Space ◽  
Plasma Membranes ◽  
Freeze Fracture ◽  
Septate Junction ◽  
Fracture Face ◽  
Septate Junctions ◽  
Thin Sections ◽  
Intramembranous Particles ◽  
Lanthanum Tracer

A septate junction is described in reproductive pairs of the protozoon Gregarina polymorpha, using conventional thin sections, lanthanum tracer and freeze-fracture techniques. The septate junction is established between the plasma membranes at the tips of the joined epicytic folds. It is characterized by an intercellular space of 14–17 nm traversed by septa with a repeat of 15–25 nm. Lanthanum-treated material exhibits transparent curves forming a meshwork. Freeze-fracture replicas show membrane modifications in the shape of short rows of intramembranous particles on the E fracture face of the plasma membrane. The significance of the finding of such a septate junction between protozoan cells is discussed.

Phylogenetic Relationships within Theinvertebrata in Relation to Thestructure of Septate Junctions and the development of ‘Occluding’ Junctional Types

1982 ◽  
Vol 53 (1) ◽  
pp. 279-305 ◽  
Author(s):  
COLIN R. GREEN ◽  
PATRICIA R. BERGQUIST
Keyword(s):  
Tight Junction ◽  
Phylogenetic Relationships ◽  
Freeze Fracture ◽  
Intercellular Junctions ◽  
Septate Junction ◽  
Simple Type ◽  
Septate Junctions ◽  
Exterior Surface ◽  
Lanthanum Tracer ◽  
To Come

The structures of 13 variants of invertebrate septate junction are reviewed on the basis offreeze-fracture, lanthanum tracer and thin-section studies. In addition, a simple type ofoccluding junction in the phylum Porifera, a variation of tight junction in the phylum Tunicateand the vertebrate tight junction are covered. All the junctions considered form a belt around the apical circumference of cells lining a lumen or an exterior surface. The large number of these junctions now recognized permits discussion relating to invertebrate classification and suggested phylogenetic relationships, and to the development of intercellular junctions. The relationships revealed are discussed under three headings: Coelenterates and lower invertebrates, Proterostomia (the annelid, molluscan and arthropod lineage) and the Deuterostomia(the echinoderm and chordate lineage). It is proposed that the pleated septate junction of the lower invertebrates resembles that of the hydrozoan rather than anthozoan Coelenterates. This lower invertebrate pleated septate junction occurs in several lower invertebrate phyla including the Annelida (of the proterostome lineage), but also occurs in the Sipunculoidea, a group supposedly on the deuterostome lineage.The proterostome line includes the molluscs and the arthropods, which have the molluscarthropodpleated septate junction. Several variations of the smooth septate junction are alsoseen in Arthropoda. Among the deuterostomes the Chaetognatha have both a paired septatejunction and a pleated junction and are therefore considered to be not very far removed fromthe Sipunculoidea. The echinoderms and hemichordates also have double-septum septatejunctions. In addition however, these two phyla have anastomosing septate junctions thatare very similar, varying only in their final configuration. Of the two, the echinoderm anastomosingseptate junction most closely resembles the tight junction seen in the tunicates, and the Hemichordata are therefore considered to be a lateral development from the main lineof chordate evolution. The tunicates have a tight junction similar to that seen in vertebrates;it is however more ‘leaky’ and has distinctive freeze-fracture characteristics.In the phylum Porifera a form of simple parallel membrane junction appears to serve anoccluding function. This junction has regular intercellular spacing in the absence of any septaand it is suggested that the spacing in septate junctions is probably not dictated by the septa.This interpretation is reasonable particularly when the diversity of septal types in conjunctionwith stable intercellular spacing is considered. Finally, a theory is put forward suggesting thatin evolution a change from the septate to the tight junction could simply involve a modificationof a ‘membrane spacing factor’, which allows the membranes of adjacent cells to come together at intervals, in the normal tight junction pattern.

scholarly journals Septate junctions in imaginal disks of Drosophila: a model for the redistribution of septa during cell rearrangement.

1982 ◽  
Vol 94 (1) ◽  
pp. 77-87 ◽  
Author(s):  
D K Fristrom
Keyword(s):  
Freeze Fracture ◽  
Septate Junction ◽  
Lateral Boundary ◽  
Progressive Change ◽  
Septate Junctions ◽  
Thin Sections ◽  
Cell Rearrangement ◽  
Imaginal Disks ◽  
Discrete Domains ◽  
Small Channels

The organization of septate junctions during morphogenesis of imaginal disks is described from freeze-fracture replicas and thin sections with a view to understanding junction modulation during rearrangements of cells in epithelia. The septate junctions of each epithelial cell of the disk are distributed in a number of discrete domains equal to the number of neighboring cells. Individual septa traverse domains of contact between pairs of adjacent cells, turn downwards at the lateral boundary of the domain and run parallel to the intersection with a third cell. This arrangement leaves small channels at three-cell intersections that are occupied by specialized structures termed "tricellular plugs." Cell rearrangement involves a progressive change in the width of contact domains between adjacent cells, until old contacts are broken and new ones established. It is proposed that the septate junction adjusts to the changing width of domains by the compaction or extension of existing septa. This redistribution of septa theoretically allows a transepithelial barrier to be maintained during cell rearrangements. The applicability of this model to other epithelial tissues is discussed.

scholarly journals Development of cell junctions in sea-urchin embryos

1983 ◽  
Vol 62 (1) ◽  
pp. 27-48
Author(s):  
E. Spiegel ◽  
L. Howard
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Digestive Tract ◽  
Sea Urchin ◽  
Freeze Fracture ◽  
Cell Junctions ◽  
Septate Junctions ◽  
Thin Sections ◽  
Sea Urchin Embryos ◽  
Lanthanum Tracer

The development of cell junctions in sea-urchin embryos has been investigated using thin sections, lanthanum-tracer and freeze-fracture techniques. Three types of desmosomes are present: belt desmosomes and spot desmosomes, which attach cells to each other, and hemi-desmosomes, which attach cells to the basement membrane. Two types of septate junctions are present: the straight, unbranched, double-septum septate, which is present in epithelial cells throughout embryogenesis, and the pleated, anastomosing, single-septum septate. The latter is formed only on cells that have invaginated to the interior of the embryo to form the digestive tract. The pleated junctions are shown to replace the straight junctions that were originally present before the cells migrated to the interior. It is suggested that these pleated septates may be specialized for digestive processes, since they are developed just prior to feeding and are retained in the adult intestine. Tricellular junctions, which join the bicellular junctions of three adjoining cells, have been identified in the embryo and in the adult intestine. Evidence for the presence of gap junctions was not obtained, but there are indications of their presence.

Variations of separate junction structure in the invertebrates

1978 ◽  
Vol 36 (2) ◽  
pp. 338-339
Author(s):  
Colin R. Green
Keyword(s):  
Thin Section ◽  
Freeze Fracture ◽  
Epidermal Cells ◽  
Septate Junction ◽  
Septate Junctions ◽  
Wide Range ◽  
Lanthanum Tracer ◽  
Endodermal Cells ◽  
Junction Structure

Three main variations of the invertebrate septate junction are now generally accepted; the Hydra type, the pleated septate and the smooth septate junctions. A junctional study of many members of a wide range of invertebrate phyla using thin section, lanthanum tracer and freeze-fracture techniques has however revealed at least eight distinct septate junction types, including two anastomosing septate junctions in the higher invertebrate phyla.In the Coelenterata three forms of septate junction occur. The Hydra type found in Hydrozoa (Fig 1), a pegged junction seen in the epidermal cells of Anthozoa and a ladder-like junction seen in the endodermal cells of Anthozoa. The pegged Anthozoa junction consists of septa with distinct short pegs branching at right angles mainly from one side (fig 2). Where two septa run close together, the pegs may form crossbars linking them. The ladder junction has a pegged double septum with crossbars linking the two parts of each septum (fig 3).

scholarly journals COEXISTENCE OF GAP AND SEPTATE JUNCTIONS IN AN INVERTEBRATE EPITHELIUM

1971 ◽  
Vol 50 (1) ◽  
pp. 92-101 ◽  
Author(s):  
A. J. Hudspeth ◽  
J. P. Revel
Keyword(s):  
Electron Microscope ◽  
Gap Junctions ◽  
Gap Junction ◽  
Extracellular Space ◽  
Intercellular Junctions ◽  
Septate Junction ◽  
High Electron ◽  
Septate Junctions ◽  
Electrotonic Coupling ◽  
Lanthanum Tracer

The intercellular junctions of the epithelium lining the hepatic caecum of Daphnia were examined. Electron microscope investigations involved both conventionally fixed material and tissue exposed to a lanthanum tracer of the extracellular space. Both septate junctions and gap junctions occur between the cells studied. The septate junctions lie apically and resemble those commonly discerned between cells of other invertebrates. They are atypical in that the high electron opacity of the extracellular space obscures septa in routine preparations. The gap junctions are characterized by a uniform 30 A space between apposed cell membranes. Lanthanum treatment of gap junctions reveals an array of particles of 95 A diameter and 120 A separation lying in the plane of the junction. As this pattern closely resembles that described previously in vertebrates, it appears that the gap junction is phylogenetically widespread. In view of evidence that the gap junction mediates intercellular electrotonic coupling, the assignment of a coupling role to other junctions, notably the septate junction, must be questioned wherever these junctions coexist.

scholarly journals Definitive Evidence for the existence of tight junctions in invertebrates

1980 ◽  
Vol 86 (3) ◽  
pp. 765-774 ◽  
Author(s):  
NJ Lane ◽  
HJ Chandler
Keyword(s):  
Tight Junctions ◽  
Freeze Fracture ◽  
Permeability Barrier ◽  
Thin Sections ◽  
Definitive Evidence ◽  
Common Precursor ◽  
En Face ◽  
Spider Central Nervous System ◽  
Exogenous Compounds ◽  
Colloidal Lanthanum

Extensive and unequivocal tight junctions are here reported between the lateral borders of the cellular layer that circumscribes the arachnid (spider) central nervous system. This account details the features of these structures, which form a beltlike reticulum that is more complex than the simple linear tight junctions hitherto found in invertebrate tissues and which bear many of the characteristics of vertebrate zonulae occludentes. We also provide evidence that these junctions form the basis of a permeability barrier to exogenous compounds. In thin sections, the tight junctions are identifiable as punctate points of membrane apposition; they are seen to exclude the stain and appear as election- lucent moniliform strands along the lines of membrane fusion in en face views of uranyl-calcium-treated tissues. In freeze-fracture replicas, the regions of close membrane apposition exhibit P-face (PF) ridges and complementary E-face (EF) furrows that are coincident across face transitions, although slightly offset with respect to one another. The free inward diffusion of both ionic and colloidal lanthanum is inhibited by these punctate tight junctions so that they appear to form the basis of a circumferential blood-brain barrier. These results support the contention that tight junctions exist in the tissues of the invertebrata in spite of earlier suggestions that (a) they are unique to vertebrates and (b) septate junctions are the equivalent invertebrate occluding structure. The component tight junctional 8- to 10-nm-particulate PF ridges are intimately intercalated with, but clearly distinct from, inverted gap junctions possessing the 13-nm EF particles typical of arthropods. Hence, no confusion can occur as to which particles belong to each of the two junctional types, as commonly happens with vertebrate tissues, especially in the analysis of developing junctions. Indeed, their coexistance in this way supports the idea, over which there has been some controversy, that the intramembrane particles making up these two junctional types must be quite distinct entities rather than products of a common precursor.

Isolation and characterization of invertebrate smooth septate junctions

1983 ◽  
Vol 62 (1) ◽  
pp. 351-370
Author(s):  
C.R. Green ◽  
C. Noirot-Timothee ◽  
C. Noirot
Keyword(s):  
Molecular Weight ◽  
Gap Junctions ◽  
Freeze Fracture ◽  
Intercellular Junctions ◽  
Septate Junction ◽  
Major Protein ◽  
Septate Junctions ◽  
Isolation And Characterization ◽  
Significant Difference ◽  
Freeze Fracture Electron Microscopy

Using modifications of techniques used for the isolation of macula type intercellular junctions (gap junctions and desmosomes) the arthropod smooth septate junction has been isolated from insect midgut tissue. Midguts from cockroaches or mealworms were used and membrane fractions were obtained by sucrose gradient and ultracentrifugation techniques. Preparations with reasonable concentrations of septate junction were obtained and have been studied by thin-section, negative-stain and freeze-fracture electron microscopy. The junctions appeared to be well preserved, although there was evidence that the junction strands were able to slide within the plane of the membrane. Septa were seen to have a cross-striated appearance when viewed after negative staining but their exact structure remained difficult to determine. Polyacrylamide gel electrophoretic studies demonstrated the reproducibility of the isolation procedure and showed that septa may have a 47 000 molecular weight glycoprotein component. Gel electrophoresis also gave some indication of the intramembrane biochemistry of the smooth septate junction, with proteins of 31 000 and 32 000 molecular weight always occurring in the junction fractions. The junctions were, however, very sensitive to both mechanical and chemical treatments, the septa were destroyed by rough homogenization or by treatment with urea at a concentration as low as 1 M. Freeze-fracture of untreated, isolated junctions demonstrated no differences from junctions in intact tissue, while replicas of urea-treated material were more difficult to interpret as the component parts of the junctions became separated once the septa had been destroyed. Gap junctions were also obtained and resisted both mechanical and chemical treatment, which destroyed the septate junctions. Their major protein component appeared to have a molecular weight of 36 000. Attempts to isolate pleated septate junctions (from insects, molluscs and annelids) by the same techniques failed, implying a significant difference in the structures of the two types of septate junction.

Two new septate junctions in the phylum Coelenterata

1980 ◽  
Vol 42 (1) ◽  
pp. 43-59
Author(s):  
C.R. Green ◽  
N.E. Flower
Keyword(s):  
Particle Size ◽  
Freeze Fracture ◽  
Outer Edge ◽  
The Other ◽  
Septate Junction ◽  
Epithelial Tissue ◽  
Fixed Tissue ◽  
Septate Junctions ◽  
Lanthanum Tracer ◽  
Endothelial Tissue

Freeze-fracture of fixed and unfixed tissue, lanthanum tracer and conventional thin-section studies have revealed 2 new types of septate junction in the class Anthozoa, phylum Coelenterata. These new junctions have the 15–18-nm intercellular spacing of all other described septate junctions and are found around the apical circumference of cells lining a lumen or outside edge. However, in freeze-fracture replicas and tangential views of lanthanum-impregnated tissue, they are seen to be quite different from other known septate junction types. One of the new junctions is found in endothelial tissue such as that lining the gut or the inside of the tentacles. In tangential view it is seen to consist of relatively short, straight, double septa, again with lateral projections. In feeeze-fracture of unfixed tissue, the junction consists of double rows of particles on the P face, the particles of one row being rounded, those of the other being elongated at right angles to the line of the septum. This dichotomy in particle size is unexpected, as the 2 halves of the septa as seen in tangential view are symmetrical. In freeze-fracture of fixed material the particle arrays remain on the P face and appear similar to those of unfixed material, but never as clear. In fixed tissue, some distortion had occurred and in extreme cases septa appear as a single broad jumbled row of particles. In this double septa junction, the rows of particles seen in freeze-fracture are occasionally seen to anastomose with a septum dividing into 2 and a third row of particles aligning with the 2 new septa to form their double particle rows. In both fixed and unfixed tissues, the E face of the junction consists of wide, shallow grooves. The second of the new junctions occurs in epithelial tissue, such as around the outer edge of sea-anemone tentacles, and consists of long wavy septa with lateral projections. In views where these projections appear longest, they arise predominantly from one side of the septa. In freeze-fracture of both fixed and unfixed tissue, this junction appears as rows of closely spaced particles on the P face. Occasionally rows of particles are seen on the E face, but usually this face is characterized by shallow grooves. In some aspects these 2 new junctions have features in common with the Hydra type junction also found in the Coelenterata. In all 3 types septa are relatively straight, rather than pleated, and there are lateral projections on the septa.

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.

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