Typical Gap Junctions Research Articles

Membrane junctions on odontoblasts

The morphology of the odontoblastic cell layer in the human dental pulp was analysed using freeze-fracturing technique. Numerous intercellular junctions were found between individual odontoblasts as well as between odontoblasts and subodontoblastic cells (Hohl's cells). These junctions were of the gap junction type. Tight junctions were not observed.

Gap junctions in laryngeal carcinoma.

The ultrastructural morphology of gap junctions in malignant keratinocytes of invasive squamous cell carcinoma of the human larynx was studied by electron microscopy. The ultrastructure of gap junctions in the malignant keratinocytes was similar to the typical intercellular gap junctions described in normal tissues. In human laryngeal carcinoma, however, the gap junctions were noticeably reduced in number as compared to normal epithelia. Spherical intracytoplasmic structures limited by gap junctional membranes were also found. These annular gap junctions arise by invagination of the cell surface membrane. The loss of gap junctions in laryngeal carcinoma is probably one of the many prerequisites which facilitate tumor invasion.

Gap junctions in early amphibian embryos

Cell pairs from late cleavage and early blastula Ambystoma mexicanum embryos were found to be electrotonically coupled. Thin-section electron microscopy revealed typical gap junctions between the cells. Freeze-fracture electron microscopy showed the gap junctions to be comprised of aggregations of 8.5-nm P-face particles and corresponding E-face pits. The conductance of the amphibian embryonic gap junction is known to be voltage-dependent, but no obvious gating structure was resolved by these morphological techniques.

A Freeze-Etching Study

In order to investigate the myelin and the glial cell membranes in the optic nerve of the mutant mouse "Jimpy," the method of freeze-etching was applied. The compact myelin lamellae and the first two glial membranes of the mutant, as compared to the normal mouse, show several abnormalities: absence of intramembraneous particles on the P-face, myelin lamellae separated by cytoplasmic layers, vesicular protrusions forming irregular invaginations, and elevations and tight junctions with a discontinuous, zigzag course. Some of these characteristics were found in the membrane of the oligodendroglia cell of the pathological animal, as well. The astrocytic membranes of both normal and Jimpy mice contain two types of gap junctions. The occurrence of one type seems to be increased in the mutant. Freeze-fractured internodal regions of the axolemma are not significantly different from those of normal animals.

Toad urinary bladder epithelial cells in culture: maintenance of epithelial structure, sodium transport, and response to hormones.

Epithelial cells from the toad urinary bladder have been grown in continuous culture. Many of the cells resemble the granular cell type of the urinary bladder. They form an epithelium with typical tight junctions and gap junctions. The transport properties of two cell lines have been examined. When cells of the line designated TB-M or of line TB-6c are grown on collagen-coated nucleopore filters, epithelia are formed that have transepithelial potential differences of 40 and 20 mV, resistances of 5000 and 10,000 omega-cm2, and short-circuit currents (ISC) of 8.5 and 2.5 muA/cm2, respectively. Net mucosa to serosa sodium transport accounts for all of ISC in line TB-M and for 70% of ISC in line TB-6c. Vasopressin, which stimulates adenylate cylase and ISC in the intact bladder, has no effect on the cells in culture. Cyclic AMP stimulates ISC and lowers resistance in both lines. Aldosterone stimulates ISC in both lines. This is accompanied by a fall in resistance in line TB-M and no change in resistance in line TB-6c. Amiloride inhibits ISC in TB-M cells under basal conditions and after stimulation by aldosterone. In line TB-6c amiloride has no effect under basal conditions but lowers ISC of aldosterone-treated cells to the basal level. Thus, the cells have retained the ability to form oriented, high-resistance epithelial membranes that manifest hormone-sensitive transepithelial sodium transport.

Development of Sertoli cell junctions in vitro--a freeze-fracture study.

Seminiferous tubules of 1-day-old rats were maintained in organ culture for up to 40 days. Five classes of intercellular junctions between Sertoli cells were observed by the freeze-fracture method as the tissue aged: (a) typical gap junctions; (b) focal tight junctions; (c) macular tight junctions; (d) meandering tight junctions; and (e) extensive tight junctions. The relative proportions of these types of Sertoli cell junctions were quantitated as the organ cultures progressed. The junctional structures observed and classified in organ culture were identical to those seen in vivo, but the timing of their appearance and/or disappearance, as well as their relative proportions, was different from that observed in the developing animal. Extensive tight junctions, with numerous parallel strands, were observed in the 40-day cultures; however, their oblique orientation with respect to the myoid layer was in contrast to the parallel orientation observed in vivo.

Mixed (chemical and electrical) synapses on frog spinal motoneurons.

Freeze-fracture replicas and ultrathin sections were used to characterize the gap junctions on the somata and large dendrites of frog motoneurons found earlier by Sonnhof et al. (1977). In freeze-fracture replicas one of the specific features of these relatively frequent gap junctions is the presence of circular regions of non-junctional membrane ("fenestrae") within areas of typical gap junction appearance displaying P-face particles or E-face pits. Such "fenestrated" gap junctions are mostly associated with membrane specializations indicative of the active zone of a chemical synapse (including vesicle attachment sites in non anaesthetized animals) to constitute mixed synapses. These findings could be verified in ultrathin sections, which revealed that the vesicles of the chemical component of the mixed synapses were spherical and agranular. Our results suggest that the mixed synapses are predominantly axo-somatic and axo-dendritic. The existence of dendro-dendritic gap junctions in the ventral horn region as described by Sotelo and Taxi (1970) was verified in ultrathin sections; they were rare, solely electrotonic in character, and probably represent the morphological basis for the VR-EPSP (Katz and Miledi, 1963; Kubota and Brookhart, 1963), i.e. electrotonic coupling between motoneurons of different spinal segments (Washizu, 1960). Electrotonic coupling can also be demonstrated between motoneurons and afferent fibers of the dorsal root and the lateral column. Electrotonic potentials recorded within motoneurons during electrical stimulation of dorsal root or lateral column precede the chemical postsynaptic potentials; after Mn2+-blockade of chemical synaptic transmission, the electrotonic component persists. Some fibers of these afferent pathways are therefore assumed to act monosynaptic on the motoneuron via mixed axo-somatic and axo-dendritic synapses.

Junctional complexes and variations in gap junctions between spinal cord ependymal cells of a teleost, Sternarchus albifrons (gymnotoidei)

Junctions between ependymal cells lining the central canal in the spinal cord of the weakly electric teleost, Sternarchus albifrons, were investigated by means of thin-section and freeze-fracture electron microscopy. Junctional complexes representing the ‘terminal bar’ consist of a zonula occludens, followed by intermediate junctions and desmososomes. This pattern is common in many types of epithelia but uncommon in ependyma. Three types of gap junctions were found below the leel of the terminal bar and intermixed with desmosomes: (1) gap junctions at least partially enclosed by a single strand of apparent tight junction, (2) ordinary gap junctions, partially surrounded by a particle-free halo and (3)_segmented gap junctions, in which linear arrays several particles wide are separated by fairly regular linear particle-free regions. Some of the observations may have implications for the mechanism of formation of gap junctions.

Freeze-etching observation on the development of intercellular junctions of the duodenal epithelial cells in the chick embryo.

Development of intercellular junctions in duodenal epithelium in the chick embryo was studied by electron microscopy using the thin-section and freeze-fracture techniques. Incomplete tight junctions are already seen in 6 and 7 day old embryos at the apical portion of the lateral plasma membrance, and consist of 1-7 strands, their mean depth measuring 0.2 micron. This corresponds to a "very leaky type" (CLAUDE and GOODENOUGH) of tight junction. Ridges on the PF are discontinuous and rarely cross or link. The tight junctions extend basally at the place where more than three epithelial cells are in contact. On the lateral plasma membrane, particle-aggregates suggesting primitive gap junctions are already recognized. Some are dense aggregations of 3-5 membrane-particles with a halo free of the particles and others are rather loose aggregations of 5-10 particles with an indistinct halo. In 9 day old embryos, the ridges of the tight junction become more discontinuous, although the frequency of the linkage of the neighboring ridges increases. The compartments bounded by the tight junctional strands are angular. These strands become continuous and the facets surrounded by them are roundish in 12 day old embryos. The presumptive immature gap junctions show a characteristic polygonal pattern in 9 day old embryos and gradually increase in size. Mature tight junctions and typical gap junctions of 0.3-0.4 micron diameter are seen after 18 days of incubation. The strands number 6-8 and the depth of the tight junction measures about 0.4 micron in 18 day old embryos. In the chick embryo duodenal epithelium, the tight and gap junctions develop independently from each other without any direct interaction between them.

Intercellular junction development in maturing rat seminiferous tubules

Sertoli cells of the seminiferous tubules in immature rats (1–20 days) contain gap and tight junctions in different stages of development as identified in freeze-fracture replicas. Typical gap junctions and gap junction formation were observed between Sertoli cells. Tight junctions were observed to assume a variety of configurations including linear, macular, and extensive occluding complexes. Tight junction formation was also observed. A decrease in the frequency of gap junctions and a corresponding increase in the number of tight junctions was quantitatively shown.

Freeze-fracture studies of gap junctions in the developing and adult amphibian cardiac muscle

The freeze-fracture technique has been used to characterize the junctional devices involved in the electrical coupling of Ambystoma cardiac tissue. These cells are connected by junctions formed by either linear or circular arrays of particles. Such structures can be interpreted as a special type of gap junction. Gap junctions have also been investigated during the growth and differentiation of two amphibians, Rana and Xenopus. In both genera the earliest stage of junctional assembly is characterized by linear rows of particles. Later, a gradual transformation of these linear rows into circles was found. Finally, in the fully formed gap junctions, these circles appeared to join together into clusters. In summary, in the adult amphibian myocardial cells, three different types of gap junctions can be described. The first type, which has been observed in all embryonic stages and in adults in all three genera, consists of linear or circular arrays of particles: this is the only type of gap junction seen at any age in Xenopus. The second type, consisting of a variable number of anastomosing circles forming regular networks, is never observed in embryonic cells. It is typical of the adult frog heart and may also be seen in Ambystoma. The third type is characteristic only of adult Ambystoma heart and consists of geometrically packed particles identifiable with classic communicating macula. The fact that only the first class of structure is observed in Xenopus heart strongly supports the conclusion that such linear arrays of intramembranous particles really represent true functional electrical junctions.

Intercellular junctional complexes of the rat seminiferous tubules: a freeze-fracture study.

This paper provides a description of the intercellular junctions in the rat testis, as observed using the freeze fracture technique. These intercellular junctions were categorized into four general types: Sertoli cell tight junctions, myoid cell tight junctions, gap junctions, and "heterogeneous junctions." The Sertoli cell tight junctions had a mean depth of 3.1 micron in the basal to apical direction, and contained 25 to 50 (average: 36) parallel rows of particulate sealing elements. The myoid cell tight junction was neither continuous nor extensive, but focal in nature. Interestingly the sealing elements of this junction, like those of the Sertoli cell tight junction, were quite particulate in nature. Typical gap junctions were observed between Sertoli cells where they were intercalated between the parallel rows of the Sertoli cell tight junction. The most interesting observation was the identification of gap junction-like structures, in various stages of formation, on germ cell membrane fracture faces, both in the basal and adluminal compartments. Lastly, the unusual "heterogeneous junction" was observed on large membrane fracture faces in close proximity to cells in the adluminal compartment, presumably between Sertoli cells. These junctions appeared to consist of both tight and gap junction elements.

Gap junctions in the cardiac muscle cells of the lamprey.

Electron microscopy of both thin sections and freeze-fracture replicas has demonstrated the occurrence of gap junctions (nexuses) in the cardiac muscle cells of the lamprey. These gap junctions are identical in basic structure with those found in the mammalian heart. However, they are much smaller (less than 0.5 micron in diameter), and more irregularly distributed than the typical gap junction in the mammalian heart. These small gap junctions seem to provide a structural basis for the electrical coupling between cardiac muscle cells in the lower vertebrates. In addition, the well developed sarcoplasmic reticulum and subsurface cisternae, which contain an electron dense spheroidal cast, frequently observed in the cardiac muscles of the lamprey.

A decay of gap junctions in association with cell differentiation of neural retina in chick embryonic development.

Ultrastructural studies of thin-sectioned and freeze-cleaved materials were performed on developing retinal tissues of 3- to 9-day-old chick embryos to clarify the junctional structures between neural retinal cells and between neural retinal cells and cells of the pigmented epithelium. Frequency, size and position of gap junctions in developing neural retina are different at each stage of development. In 3-day-old embryos, some cells adhere to each other by gap junctions immediately below the outer limiting membrane of neural retinae. The size and number of gap junctions increase remarkably during 5-6 days of incubation. In this period of development, well developed gap junctions consisting of subcompartments of intramembrane particles are found between cell surfaces at both the outer limiting membrane region and the deeper portion of the neural retina. Gap junctions disappear thereafter, and at 7-5 days of incubation, small gap junctions are predominant between cell surfaces at the outer limiting membrane region, while the frequency of gap junctions in the deeper portion is very low. At 9 days of incubation, gap junctions are rarely found. Typical gap junctions are always found between neural retinal cells and those of the pigmented epithelium in embryos up to 7-5 days of incubation. Tight junctions are not found in the neural retina or between neural retina and pigmented epithelium throughout the stages examined.

Gap junctions in the differentiated neural retinae of newly hatched chickens.

Gap junctions in the neural retinae of newly hatched chickens were examined in thin section and by freeze cleaving. Unusual gap junctions containing linear arrays of intramembrane particles are found between principal and accessory cones which form a double cone at the region of the outer limiting membrane. These unusual gap junctions are often continuous with macular aggregates of hexagonally packed intramembrane particles which are characteristic of a typical gap junction. Typical gap junctions are also found in both the outer and the inner plexiform layers and in the outer nuclear layer, but are not so abundant as in the outer limiting membrane region. The sizes of intramembrane particles and their centre-to-centre spacing within the macular aggregate of a gap junction in differentiated neural retinae are slightly larger than those in undifferentiated neural retinae. Tight junctions are not found in differentiated neural retinae.

Lateral membrane morphology and gap junction structure in rabbit corneal endothelium

The morphology and the intercellular contacts of the lateral plasma membranes in the rabbit corneal endothelium have been observed by the lanthanum tracer technique. When the lateral cell boundaries show extensive interdigitation, they also send bulbous projections from one cell into its neighbor and gap junctions bridge the lateral plasma membranes. The basic features of the intercellular contacts are similar to the typical gap junction, having a polygonal array of subunits with a center-to-center spacing of about 90 Å bridging a narrow intercellular space. In many of the junctions observed two additional features are prominent: the subunits form linear arrays and the intercellular space within the junction shows a periodic, 320–360 Å center-to-center, widening. The latter feature appears as lanthanum-filled channels where the narrow intercellular space with the subunit array widens from 30–40 to 60–80 Å.

Junctions in developing human and rat kidney: A freeze—fracture study

The developing nephrons of human and rat kidney contain a wide spectrum of junctional specializations detectable in freeze—fracture replicas. The plasma membranes of developing podocytes show junctions which undergo changes in position and shape during the differentiation of the cells and depart morphologically from the usual “tight” type in being formed of rows of particles instead of continuous ridges on the A (or P) face. In fetal kidney tubules, tight junctions are found which closely resemble those previously described in similar segments of the adult nephron. Moreover, unidentified epithelial structures corresponding probably to less differentiated stages of the nephron present atypical junctions formed on the A face by interconnecting linear elevations or crests carrying a few particles and on the B (or E) face by patterns of grooves containing numerous adhering particles. Typical gap junctions are noticed in developing proximal tubules.

The Sertoli cell occluding junctions and gap junctions in mature and developing mammalian testis

Special occluding junctions between Sertoli cells near the base of the seminiferous epithelium are the structural basis of the blood-testis permeability barrier. In micrographs of thin sections, multiple punctate pentalaminar contacts between apposed membranes are observed in the junctional regions. In freeze-fractured mature testis, the junctional membranes exhibit up to 40 parallel circumferentially oriented rows of intramembrane particles preferentially associated with the B-fracture face, but with complementary shallow grooves on the A-face. Short rows of particles may remain with the A-face resulting in discontinuities in the B-face particle rows. In addition, elongate aggregations of particles of uniform size (∼70 A) arranged in one or more closely packed rows are occasionally found adjacent to the linear depressions on the A-face of the Sertoli junction. These are interpreted as atypical gap junctions. In immature testis, occluding junctions are absent but typical gap junctions are common. These gradually disappear. In the second postnatal week, linear arrays of particles appear on the B-face. Initially meandering and highly variable in direction, these gradually adopt a consistent orientation parallel to the cell base. The establishment of the blood-testis barrier appears to be correlated with this reorganization of the intramembrane particle rows. Sertoli junctions were shown to be resistant to hypertonic solutions that rapidly dissociate junctions of other epithelia. Sertoli junctions thus differ from other occluding junctions in their (1) basal location, (2) large number of parallel particle rows, (3) absence of anastomosis between rows, (4) preferential association of the particles with the B-face, (5) intercalation of atypical gap junctions, (6) unusual resistance to dissociation by hypertonic solutions.

Low resistance junctions between mesoderm cells during development of trunk muscles.

1. Electrical connexions between mesoderm cells have been examined during the formation of somites in Xenopus laevis, Bombina orientalis and Ambystoma mexicanum. 2. In Xenopus the resting potentials of presumptive myotome cells (-65 + 2 mV, S.E. of mean) and somite muscle cells (-65 +/- 0-6 mV S.E. of mean) were 40 mV, greater than dermatome cells (-25 +/- 0-6 mV, S.E. of mean). Similar differences were found in Bombina and Ambystoma. 3. In all three species cells of the dermatome layer of the mesoderm were electrically coupled to each other. Cells of the presumptive myotome layer in the unsegmented region of the mesoderm were also electrically coupled. 4. In Xenopus dermatome and myotome layers of the mesoderm were not electrically coupled to each other either before or after somite formation. In the other two species dermatome and myotome layers were uncoupled once the somites had formed. 5. In all three species the position of the intersomite border in the unsegmented mesoderm region is marked by the breaking of electrical contracts between those cells destined to form the next somite and the rest of the unsegmented mesoderm. 6. In the axolotl each somite remains electrically insulated from its neighbour. In Xenopus and Bombina electrical connexions are re-established between somite muscle cells once the morphogenetic movements underlying somite formation are complete. 7. Presumptive myotome cells in Xenopus and Ambystoma acquire the membrane property of inward-going rectification before incorporation into a somite. 8. Once Xenopus and Bombina embryos show spontaneous movements large end-plate potentials are recorded from the head somites. Excitation spreads from somite to somite along the low resistance intercellular pathway allowing simultaneous contraction of several somites before extensive somite innervation. 9. The structure of developing somite muscle of Xenopus has been studied with the electron microscope. 10. Close membrane contacts of the gap junction type have been seen between undifferentiated presumptive myotome cells, muscle cells in the same somite and between muscle cells in adjacent somites. 11. Myofilament organization first begins in mesoderm cells when they are forming a new somite. Complete sarcomeres appear in the head somites when the embryo begins spontaneous flexion movements.

A morphological basis for intercellular communication between alpha- and beta-cells in the endocrine pancreas.

By degranulating beta-cells in the islets of Langerhans of the rat with sulfonylurea, it has been possible to distinguish unambiguously alpha-cells from beta-cells in freeze-fracture replicas. In such preparations, we found morphologically typical tight and gap junctions occurring between alpha- and beta-cells. The presence of gap junctions offers indirect evidence that these cells are coupled with one another; coupling may influence the secretory behavior of alpha- and beta-cells maintaining glucose homeostasis within tightly constricted limits.