Intercellular junctions in the water-blood barrier of the gill lamella in the adult lamprey (Geotria australis,Lampetra fluviatilis)

Thin sections and freeze-fracture replicas of the water-blood barrier in the gill lamellae of adult lampreys (Geotria australis, Lampetra fluviatilis) demonstrate that the occluding junctions between epithelial pavement cells differ markedly from those between endothelial pillar cells in the structure and arrangement of their strands. The zonulae occludentes between pavement cells typically consist of complex networks of 4–6 strands, the mean number of which undergoes a small but significant decline when the animal is acclimated to seawater. In comparison, the occluding junctions between pillar cells are less elaborate and may represent maculae or fasciae, rather than zonulae occludentes. They do not apparently undergo a change when the animal enters saltwater. The results indicate that the main part of the paracellular diffusion barrier to proteins and ions is located in the epithelium rather than the endothelium. Communicating (gap) junctions are present between adjacent pavement cells, between pavement and basal cells and between pillar cells. These findings suggest that the epithelial cells and the pillar cells in the water-blood barrier of lampreys both form functional syncytia. The results are discussed in the context of ion-transporting epithelia in other aquatic vertebrates.This paper is dedicated to Professor H. Leonhardt on the occasion of his 75^th birthday     

Junctional diversity in two regions of the epidermis of Oikopleura dioica (Tunicata,Larvacea)

Summary A simple continuous epithelium surrounds the body of the pelagic larvacean. It consists of two zones of cells: oikoplast cells and flattened cells. The oikoplast cells are columnar and produce a thick extracellular house that ensheathes the body of the organism. These cells are joined laterally by wide tight junctions (zonulae occludentes). The tail of the animal is surrounded by exceedingly thin cells which are joined by narrow tight junctions under which lie intermediate junctions (zonulae adhaerentes) and gap junctions. A web of fibrous material inserts into the intermediate junctions. The transitional cells between the two epithelial zones have one lateral border with a wide tight junction, and the other lateral border with a narrow tight junction and a wide intermediate junction. In freeze-fracture replicas, the wide tight junction has a number of anastomosing ridges, in comparison with the narrow tight junction, which usually consists of only a single row of intramembranous particles. In replicas, the thin epithelial cells show unusual parallel arrays of particles in clusters on their apical plasma membranes. This simple epithelium, therefore, exhibits striking differences between the two cellular zones, in the structural characteristics of both the lateral borders and the apical membrane.     

“Tight” junctions in the sheath of normal and regenerating motor nerves of the crayfish,Orconectes virilis

Summary Tight or occluding intercellular junctions occur between adjacent glial processes in normal and regenerating crayfish motor nerve sheaths. Although infrequent, these junctions possess the ridge and groove configuration characteristic of freeze-cleaved occluding junctions. When present, nerve sheath tight junctions consist of a single, or at most a few, parallel intramembrane ridges situated on the EF membrane face of the glial plasma membrane. Consequently, such contacts are rarely recognized in thin sections of plasticembedded nerve sheaths. Crayfish nerve sheath tight junctions are of the fascia occludens type and, therefore, do not impede solute flow across the nerve sheath. Fasciae occludentes of regenerating nerve sheaths occur in close proximity to discoid plaque-like aggregates of particles assumed to represent maculae adhaerentes. This relationship, which was not observed in normal nerve sheaths, suggests a functional association between the two types of junctions, perhaps developmental transformation of one junction type into the other. Although ridges and grooves of tight junctions occur next to crossfractured trans-glial channels, no functional significance is proposed for this relationship. This study is the first report of tight intercellular junctions in crustacean glial nerve sheaths.Supported by the National Research Council of Canada     

The occluding junctions of mouse duodenal enterocytes during development

Summary The architecture of occluding junctions during the differentiation of the mouse duodenum was studied in freeze-fractured material. Irregular zonulae occludentes (ZO) (Type I) are numerous during fetal life, and are characterized by their irregular width, and by the presence of basal open-ended extensions fused with the discontinuous basal strand of the ZO. Regular ZOs (Type II), typical of the adult villous epithelium, appear after Type I junctions by day 16 of gestation. Two patterns are distinguishable: in the first, parallel strands of ridges and furrows are found without crossing branches; in the second pattern, the junction zone is organized like a network of short branches forming various types of polygons. In fetal and adult mice fasciae occludentes (FO) (Type III) are present on the lateral cell membranes; in unfixed specimens particles are found in the furrows of the E-face and pits on the ridges of the P-face. In fixed tissues, the particles are aligned on the ridges of the P-face. These results indicate that fixation with glutaraldehyde modifies considerably the affinity of junctional particles toward the P-face during the fracture process. Moreover, the presence of numerous large FOs on the lateral cell membranes of enterocytes during late fetal life and in the adult, is possibly related to cell movement along the intestinal villi.     

Junctions in the central nervous system of the cat

Interendothelial membrane contacts in different segments of brain blood vessels were investigated by the freeze-etching technique. The study demonstrated that the endothelial cells of the pre- and postcapillary segments were coupled by elaborate zonulae occludentes. These tight junction formations encompassed gap junctions of different sizes and distribution. The globules of the pre- and postcapillary tight junctions revealed a great fragility which led to an atypical distribution of the sealing elements. In the "typical" brain capillaries the endothelial cells were connected by continuous tight junctions. In contrast to the structural continuity of these formations the fenestrated segments of capillaries in the choroid plexus and area postrema demonstrated discontinuous fasciae occludentes. They were composed of rows of individual particles which should be regarded primarily as focal in nature.     

Occluding junctions of theNecturus gallbladder

Summary The paracellular conducting pathway of theNecturus gallbladder was studied with electrophysiological and electromicroscopic methods. The first one consists of the passage of short (5 msec) and small (32 A cm^–2) current pulses associated with a voltage scanning of the plane of the epithelium at the apical surface with a microelectrode to detect the regions where current flows. The procedure shows that (a) the conductance is evenly distributed along the intercellular regions along the intercellular spaces of the cells where occluding junctions are located; (b) the field above the occluding junctions has the shape of a bell, so that the junction can be sensed at 1–2 m from the region where the intercellular space is visualized by light microscopy; (c) the intersections between three cells, in spite of having 3 half-junctions contributing (instead of two), do not have a higher conductance than the rest of the occluding junction. Scanning electron microscopy shows that (a) cells are densely covered by microvilli which interdigitate above the region of the occluding junctions, and (b) are covered by a surface coat. With transmission electron microscopy, (a) the opening of the occluding junctions at the apical border appears irregular, and most of them oblique; (b) in the last microns the actual mouth of the junction may deviate from the course of the interspace. Freeze-fracture replicas indicate that (a) the occluding junction has a uniform width and little variations in the number of strands around the cell, except (b) at intersections between 3 cells where both, its width and the number of strands, increase toward the basal region.     

The ultrastructure of the epithelium of the ciliary body

Summary The fine structure of the human and rabbit ciliary body epithelium has been studied with the electron microscope, both under normal conditions and after paracentesis of the anterior chamber. The disposition of the junctional complexes in the two layers of the ciliary epithelium is described in detail. Junctional complexes appear particularly developed between the apical surfaces of the cells of the two layers, but are present, as in other monolayered epithelia, also between the lateral surfaces of adjacent cells of each layer. The junctional complexes connecting the apical surfaces of the cells of the two layers are represented by zonulae occludentes, zonulae adhaerentes and desmosomes following each other irregularly, with interposition of rounded dilatations of the intercellular space called ciliary channels. The zonulae occludentes and adhaerentes found along the lateral surfaces of the epithelial cells probably form discontinuous and overlapping fasciae. Moreover, the existence of a peculiar dove-tail arched junction called macula occludens is suggested. Few differences were encountered when comparing the arrangement of junctional complexes in the ciliary epithelium of man with that of the rabbit. Many desmosomes connecting the basal portion of lateral surfaces of the non-pigmented cells were found only in human ciliary bodies.The study of the modifications of the junctional apparatus of the ciliary epithelium following paracentesis of the anterior chamber, confirms the functional hypotheses on junctional complexes previously suggested: in particular only zonulae occludentes cause a real block of the intercellular spaces. On the basis of the present work, the close relationships between the number, kind and disposition of junctional complexes in epithelia and the functional possibilities of these epithelia are stressed.Dedicated to Professor W. Bargmann on his 60th birthday.Dr. Orzalesi is the recipient of a research grant from Ministero della Pubblica Istruzione for 1964.     

Different functions of tight junctions in the ascidian branchial basket

The stigmatal cells in the branchial basket of ascidians from a number of genera have been examined as to the nature and distribution of their intercellular junctions. The branchial wall consists of ciliated and parietal cells; the ciliated cells are arranged in seven rows and are associated by junctions with other cells in the same row as well as with those in adjacent rows. They are also associated by junctions with peripheral parietal cells. Junctions between adjacent ciliated cells in all cases exhibit tight junctions or zonulae occludentes. However, these cell borders also possess fasciae or zonulae adhaerentes if they are in the same row and the ciliary rootlets insert-into these junctions. If the cells are in adjacent rows they exhibit adhaerentes junctions only in species belonging to the orders Phlebobranchiata and Aplousobranchiata. In contrast, if the cells in adjacent rows belong to the order Stolidobranchiata. they never exhibit any adhaerentes junctions and the ciliary rootlets of the basal bodies from the cilia insert instead into the tight junctions and the non-junctional membrane below them. At the homologous junctional borders between adjacent parietal cells and also at heterologous junctional borders between parietal and ciliated cells, tight junctions alone occur, with no co-existing adhaerentes junctions along their lateral borders. Again, fibrils from ciliary rootlets insert into zonulae occludentes. This shows that tight junctions are capable both of forming permeability barriers, in that they can be seen to prevent the entry of exogenous tracers such as lanthanum, and of acting as adhesive devices.     

Occluding junctions in a cultured transporting epithelium: Structural and functional heterogeneity

Summary MDCK cells (epithelioid of renal origin) form monolayers which are structurally and functionally similar to transporting epithelia. One of these similarities is the ability to form occluding junctions and act as permeability barriers. This article studies the junctions of MDCK monolayers formed on a permeable and transparent support (a disk of nylon cloth coated with collagen) by combining two different approaches: (i)Scanning of the electric field: the disk is mounted as a flat sheet between two Lucite chambers and pulses of 20–50 A cm^–2 are passed across. The apical surface of the monolayer is then scanned with a microelectrode to detect those points where the current is flowing. This shows that the occluding junctions of this preparation are not homogeneous, but contain long segments of high resistance, intercalated with sites of high conductance. (ii)Freeze fracture electron microscopy: the junctions are composed of regions of eight to ten strands intercalated with others where the strands are reduced to one or two ridges. The sites of high conductance may correspond to those segments where the number of junctional strands is reduced to 1 or 2. It is concluded that the occluding junctions of MDCK monolayers are functionally and morphologically heterogeneous, with tight regions intermixed with leaky ones.     

Occluding junctions in the epithelia of the gut-associated lymphoid tissue (GALT) of the rabbit ileum and caecum

Summary The zonulae occludentes of the dome epithelia and adjacent non-dome epithelia in four locations of the gut-associated lymphoid tissue (GALT) in the rabbit ileum and caecum (Peyer's patches, sacculus rotundus, caecal lymphoid patches, appendix) were studied in freeze-fracture replicas. In all locations the zonulae occludentes of the dome epithelium are composed of more junctional strands than in the corresponding non-dome epithelium. In the dome epithelia of Peyer's and caecal lymphoid patches the zonulae occludentes show considerable structural variation; the number of superimposed strands is 10 (range 5–18). In the dome epithelia of sacculus rotundus and appendix, in addition to zonulae occludentes, extended networks of junctional strands (fasciae occludentes) are present particularly between M-cells and enterocytes. The zonulae occludentes consist of 8 to 9 (range 5–15) superimposed strands; the fasciae occludentes extend up to a depth of 20m on the lateral membranes. The presence of the fasciae occludentes correlates with the appearance of regularly shaped clusters of lymphocytes, which are most developed in the dome epithelia of sacculus rotundus and appendix. These results suggest (1) that in contrast to the dome epithelia of Peyer's and caecal lymphoid patches those of sacculus rotundus and appendix are compartmentalized, and (2) that the mobility of lymphocytes and diffusion of antigens in the dome epithelia of sacculus rotundus and appendix is restricted.     

Freeze-fracture analysis of junctional complexes in the nephron of the garter snake,Thamnophis sirtalis

Summary Zonulae occludentes are shown by freeze-fracture to be pleomorphic along the garter snake nephron. In the neck and proximal segments the occluding junctions are moderately complex with frequent discontinuities in their junctional fibrils. Junctional depth and complexity are maximal in the distal and collecting segments and discontinuities in fibrils are absent. Comparison of these results with similar observations on other tissues indicates that the zonulae occludentes in the neck and proximal segments are intermediate to leaky and that they may be very tight in the distal and collecting segments. These findings suggest that in the garter snake nephron transepithelial flow of fluid may occur primarily by passive diffusion through the zonulae occludentes in the neck and proximal segments and by cell-mediated osmotic flow in the distal and collecting segments. Gap junctions occur only in the proximal tubule and are probably involved in low resistance, intercellular movement of ions.Supported by the National Research Council of Canada. The authors wish to acknowledge the generous provision of freeze-fracture facilities by Dr. M.W. Brightman, NINDS, National Institutes of Health, Bethesda, Maryland, U.S.A.     

Isolation,characterization, and long-term culture of fetal bovine tracheal epithelial cells

Summary Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol. The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic, the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm (EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight junctions, and positive keratin staining. This study was supported in part by grants BRSG S07 RR05408-25, Biomedical Research Support Grant Program, Division of Research Resources, by ES 00159, Center Grant, National Institutes of Environmental Health Sciences, by R23-HL37621, New Investigator Award, National Heart, Lung and Blood Institutes, National Institutes of Health, and by the Health Effects Institute, an organization jointly funded by the U.S. Environmental Protection Agency (Assistance Agreement X-8120059) and automotive manufactures. The contents do not necessarily reflect the views of policies of HEI, EPA, or automotive manufacturers.     

Gap junctions are non-randomly distributed inDrosophila wing discs

Summary The distribution of gap junctions in mature larvalDrosophila melanogaster wing discs was analyzed by means of quantitative electron microscopy. Gap junctions are non-randomly distributed in the proximal-distal disc axis and in the apical-basal cell axis of the epithelium. In the epithelial cells, the surface density, number and length of gap junctions are greatest in the apical cell region and distal disc region. The average gap junction surface density is 0.0572 m^–1 and 2.77% of the lateral cell surface is composed of gap junctions. In the adepithelial cells, the gap junction surface density is 0.0005 m^–1 and 0.06% of the cell surface is composed of gap junctions. No gap junctions were observed between epithelial cells and adepithelial cells. The absolute area of gap junctions was estimated in a proximal-distal strip of cells in the disc and is considerably less in the folded regions of the epithelium compared to the flat notum and wing pouch regions. The results are discussed with respect to pattern formation and growth control in imaginal discs.     

A novel perspective: the occluding zonule encircles the apex of the Sertoli cell as observed in birds

The modulation of Sertoli cell junctions was studied in the non-seasonal rooster (Gallus domesticus) and in the seasonally breeding mallard duck (Anas platyrynchos anatidae) using thin sectioning, a junction permeability tracer, and freeze-fracture replication. During the active spermatogenic phase, the junctions of the duck appeared similar to those of the rooster, thereby establishing the duck as an avian model of seasonal modulation of Sertoli cell junctions. As with mammalian seasonal breeders, during the active phase, occluding, gap, and adhering junctions formed a junctional complex all along the long axis of the Sertoli cell. Unlike in mammals, however, no 7-nm filaments were associated with the occluding junctions. An occluding zonule encircled the Sertoli cell apico-lateral membrane domain situated above the young germ cells, and constituted a barrier to the entry of lanthanum in the basal third of the seminiferous epithelium. Toward the basal side, forming focal junctions were located on the lateral Sertoli cell membrane domain facing the young germ cells. Toward the apical side, dismantling focal junctions were located on the apical Sertoli cell membrane domain facing the older germ cells. During the duck's testicular regression, 7-nm filaments were associated with an occluding junction. In freeze-fracture replicas, each junction was formed by a continuous junctional strand that encircled the apex of the cell. The strands composed a delicate narrow meshwork: an occluding zonule. The blood-testis barrier was localized near the apex of the epithelium. The seasonal reduction in the number of the strands and the changes in their orientation did not coincide with a change in the permeability of the occluding zonule to lanthanum. In addition, the cyclic disappearance of junction-associated filaments was not correlated with a change in the permeability of the junctions but with a change in the affinity of junctional particles for one or the other fracture face. It is proposed that the Sertoli cell plasma membrane domains situated apical and basal with respect to the occluding zonule be considered apical and lateral, respectively. The remaining domain facing the basement membrane would therefore be called basal. In the duck, the occluding zonule is not seasonally shifted from the base to the apex of the Sertoli cell. Instead, it remains stationed above the younger germ cells throughout the year.(ABSTRACT TRUNCATED AT 400 WORDS)     

Intercellular junctions in the uterine epithelium of Salamandra salamandra (L.) (Amphibia,Urodela)

Summary Intercellular junctions in the uterine epithelium of the ovoviviparous urodele Salamandra salamandra were studied in pregnant and non-pregnant females by freeze-fracture technique. Junctional complexes consist of zonulae occludentes (tight junctions) and numerous maculae adhaerentes (desmosomes); z. adhaerentes and nexuses (gap junctions) could not be identified. Tight junctions are of the flexible type exhibiting loosely interconnected fibrils. The fibrillary network appears stretched more often in pregnant females possibly due to the mechanical stress of pregnancy. The structure and occurrence of the junctions identified, especially that of the tight junctions, is discussed with regard to the functions of the uterus during pregnancy.Zusammenfassung Mit Hilfe der Gefrierbruchtechnik wurden im Uterus-epithel trächtiger und nichtträchtiger Feuersalamanderweibchen (Salamandra salamandra) Zonulae occludentes und Maculae adhaerentes, jedoch keine Z. adhaerentes sowie Nexus identifiziert. Die Z. occludentes sind flexibel. Ihr fibrilläres Netzwerk ist bei trächtigen Weibchen öfter gestreckt; das ist möglicherweise auf die stärkere Ausdehnung des Uterusgewebes während der Trächtigkeit zurückzuführen. Das Vorkommen der verschiedenen Kontakt-strukturen, namentlich das der Z. occludentes, wird im Hinblick auf die Funktionen des Uterus während der Trächtigkeit diskutiert.We are indebted to Mrs. K. Ott for excellent technical assistance and to Miss Dr. U. Beigel for linguistic help     

Freeze-fracture and morphometric analysis of occluding junctions in rectal glands of elasmobranch fish

Summary The structure of occluding junctions in secretory and ductal epithelium of salt-secreting rectal glands from two species of elasmobranch fish, the spiny dogfishSqualus acanthias and the stingrayDasyatis sabina, was examined by thin-section and freeze-fracture electron microscopy. In both species, occluding junctions between secretory cells are shallow in their apical to basal extent and are characterized by closely juxtaposed parallel strands. Average strand number in the dogfish was 3.5±0.2 with a mean depth of 56±5 nm; in the stingray a mean of 2.0±0.2 strands encompassed an average depth of 18±3 nm. In contrast, the linear extent of these junctions was remarkably large due to the intermeshing of the narrow apices of the secretory cells to form the tubular lumen. Morphometric analysis gave values of 66.8±2.5 and 74.9±4.6 m/cm² for the length of junction per unit of luminal surface area in the dogfish and stingray, respectively. This junctional morphology is similar to that generally described for leaky epithelia. In comparison, the stratified ductal epithelium which carries the NaCl-rich secretion to the intestine is characterized by extensive occluding junctions which extend 0.6–0.8 m in depth and consist of a mean of 12 strands arranged in an anastomosing network, an architectural pattern typical of tight epithelia. The length density of these junctions in the dogfish rectal gland was 7.6±0.1 m/cm².The junctional architecture of the rectal gland secretory epithelium (few strands, large junctional length densities) is similar to that described for several other hypertonic secretory epithelia [20, 34] and is compatible with the recent model for salt secretion in rectal glands [39] and in other Cl^– secretory epithelia which posits a conductive paracellular pathway for transepithelial Na⁺ secretion from intercellular space to the lumen to form the NaCl-rich secretory product.     

Gap junctions between pinealocytes

Summary The intercellular junctions between the pinealocytes of male rats were investigated by freeze-fracture and conventional electron microscopy.Our findings reveal that the intercellular contacts between pineal cells, formerly described as zonulae adhaerentes or zonulae occludentes, are in fact gap junctions which are difficult to characterize in thin sections due to their peculiar geometrical arrangement, which is in the form of fenestrated communicating zonules.The arrangement of these communicating zonules around rudimentary lumina of pineal clusters and rare transitions between tight and gap junctions may point to phylogenetic transformations of occluding into communicating zonules, corresponding with the change of the pineal gland from a sensory to a secretory organ. Alternatively, these tight-to-gap junctional transitions may reflect the periodic (circadian or seasonal) activity of the pineal gland.These Studies were supported by the German Research Foundation within the SFB 90 Cardiovasculäres System     

Effects of extracellular calcium depletion on membrane topography and occluding junctions of mammary epithelial cells in culture

Ca2+ dependence of occluding junction structure and permeability, well documented in explanted or cultured epithelial sheets, presumably reflects inherent control mechanisms. As an approach to identification of these mechanisms, we induced disassembly of zonulae occludentes in confluent monolayers of mouse mammary epithelial cells by exposure to low concentrations of the chelators, EGTA or sodium citrate. Stages in disassembly were monitored during treatment by phase-contrast microscopy and prepared for transmission and scanning electron microscopy. Cellular response included several events affecting occluding junctions: (a) Centripetal cytoplasmic contraction created tension on junction membranes and displaced intramembrane strands along lines determined by the axis of tension. (b) Destabilization of junction position, probably through increased membrane fluidity, augmented tension-induced movement of strands, resulting in fragmentation of the junction belt. (c) Active ruffling and retraction of freed peripheral membranes remodeled cell borders to produce many filopodia, distally attached by occluding-junction fragments to neighboring cell membranes. Filopodia generally persisted until mechanically ruptured, when endocytosis of the junction and adhering cytoplasmic bleb ensued. Junction disassembly thus resulted from mechanical tensions generated by initial centripetal contraction and subsequent peripheral cytoskeletal activity, combined with destabilization of the junction's intramembrane strand pattern.     

A new occludens-like junction linking endothelial cells of small capillaries (probably venules) of rat jejunum.

Freeze-cleave replicas of small capillaries of rat jejunum have revealed the presence of a new type of junction linking endothelial cells. This new junction reveals tight junctions (zonulae occludentes) in that the adjacent plasma membranes are held closely together along lines of attachment organized in the form of a loose, but frequently discontinuous network. In contrast to tight junctions, the A-face ridges possess a very low profile, and only at low shadowing angles can a repeating, particulate substructure occasionally be resolved. The shallow B-face furrows lack any particulate components. Images of cross-fractured focal points of attachment suggest that the external leaflets of adjacent membranes are closely apposed but not actually fused, as is the case with zonulae occludentes. It appears that this new type of endothelial junction is characteristic of small venules. Thus we propose that it be termed small venule endothelial junction.     

Cell to cell relationships in the seminiferous epithelium in the mouse embryo

Summary Germ cells and Sertoli cells in embryonic mouse testes (day 14 to 20 of gestation) were examined by sectioning and freeze-fracture. Intercellular cytoplasmic bridges between the germ cells are observed in day 14 and older embryos. Membrane specializations with dense fuzzy material similar to the socalled desmosome-like structures are found between Sertoli cells and germ cells. A cell contact area with dense opposed membranes is also found between adjacent germ cells. Asymmetrical dense fuzzy lining of both Sertoli and germ cell membranes is noted. Pinocytotic pits or caveolae are frequently found in the Sertoli cell membrane. Between adjacent Sertoli cells, gap junctions of various sizes and focal meshworks of the occluding junctions are found. Most of the occluding junctional particles are located in the center of the grooves in the E face, and are similar to those in postnatal and adult Sertoli cell junctions. In addition, on both fractured faces there are ridges and grooves devoid of particles which are continuous with occluding junctions with particles, suggesting an initial stage in the formation of occluding junctions of the Sertoli cells. Particles gathered at the site of desmosome-like structures are present on the P face of the Sertoli cell.This work is supported by the Japanese Ministry of Education