Area Of Gap Junctions Research Articles

Appearance of gap junctions in the myometrium of women during labor

Myometrial tissues from 29 women who underwent cesarean section in various stages of labor were examined quantitatively in the electron microscope. Gap junctions were present between smooth muscle cells with low frequency in women with a closed cervix and infrequent contractions of the uterus. There was a statistically significant correlation between increased cervical dilation or increased frequency of uterine contractions and increased area of gap junctions. Tissues from women in premature labor had more gap junctions than tissues from other women who underwent cesarean section either prematurely or at term. We propose that the formation of gap junctions may be stimulated by some physiologic change. The appearance of gap junctions may then terminate pregnancy by resulting in coordinated, synchronized muscle activity and dilatation of the cervix followed by expulsion of the fetus.

Tetraethylammonium modifies gap junctions between pancreatic beta-cells.

Gap junctions between pancreatic beta-cells were quantitatively assessed in freeze-fracture replicas of isolated rat islets of Langerhans incubated for 90 min with or without the potassium conductance blocker tetraethylammonium (TEA). The results show that TEA increases the median number of particles per beta-cell gap junction but not the frequency of gap junctions at both nonstimulating and threshold-stimulating concentrations of glucose. TEA increased the relative gap junctional area at both concentrations of glucose. TEA had no effect on insulin release at a basal concentration of glucose but potentiated that release at the threshold glucose level. Thus TEA modifies beta-cell gap junctions independently of its effect on insulin release. However, the junctional changes observed were greater when insulin release was also elevated.

Incidence of gap junctions and microvilli in variant cell lines with altered capacity for metabolic co-operation

The metabolic cooperation-defective embryonal carcinoma cell variant R5/3 has a reduced area of gap junctions per unit cell volume and an increased surface area of microvilli per unit volume compared with its parent PC13TG8. Both changes are reversed in the cooperation-competent revertant H2T12. This provides strong evidence in support of the view that metabolic cooperation is mediated by gap junctions, and suggests that the altered properties of R5/3 may be due to a cytoskeletal defect.

Quantitative evaluation of gap junctions during development of the brown adipose tissue.

Gap junctions of rat brown adipose tissue (BAT) were studied with the freeze-fracture technique during development. The frequency and the mean area of gap junctions increased after birth, reached a maximal development during the functional phase of the tissue (birth to 3--4 weeks) and decreased while brown adipose tissue's function regressed (from 3--4 weeks on). Gap junctional area per cell volume, an indirect estimate of intercellular coupling, followed closely the phases of BAT activity, suggesting an involvement of these junctions in the functioning of brown adipose tissue.

Fine structural differentiation of germ layers in the mouse at the time of mesoderm formation

The morphology of early postimplantation mouse egg cylinders was studied using light and electron microscopy. Implantation sites at seven, seven and one-half and eight days of gestation were dissected from the myometrium and whole implants, including both decidua and egg cylinders were processed for electron microscopy. Pre-primitive streak egg cylinders were composed of two germ layers, a tall columnar ectoderm and an outer visceral endodermal layer. Ectodermal cells demonstrated large oval nuclei and an organelle sparse cytoplasm except for many free polyribosomes. The visceral endodermal layer was composed of two cell populations. One visceral endodermal cell type observed was tall columnar in shape and appeared absorptive as demonstrated by many microvilli, pinocytotic profiles and lysosomal granules. This population was confined to extraembryonic regions of the egg cylinder. The second visceral endodermal cell type, squamous in shape, evidenced only a few microvilli, pinocytotic profiles and lysosomal granules. This population was confined to the embryonic region of the egg cylinder. Concurrent with the formation of the primitive streak an increased number of cellular junctions and nuclear pores became evident in the ectoderm. Mesodermal cells were large and stellate-shaped exhibiting many filapodia which made contact with adjacent mesodermal elements. Later the cephalic region of the primitive streak proliferated resulting in the migration of wedge-shaped mass of cells, the head process. At the most ventral extremity of the post-primitive streak egg cylinder the cells of the head process became intimately associated with the ectoderm by areas of focal contact and gap junctions.

Improved stereological techniques for studying myocardial cell growth: application to external sarcolemma, T system, and intercalated disks of rabbit and rat hearts

Stereological methods have been developed for the electron microscopic description of the growth of myocardial cells in the longitudinal and radial directions. The measurements are either independent of the contractile state or can be normalized to a reference sarcomere length. The description yields two convenient parameters, D std , an equivalent myocardial cell diameter, and L cell , an equivalent myocardial cell length expressed in dimensionless units of sarcomeres. The description takes into account sarcolemmal surface amplification due to periodic banding or the occurrence of hemispherical protuberances. Methods are also introduced for more precise estimates of the membrane areas of external sarcolemma, T system, and gap junctions. These methods have been applied to myocardial cells of rabbit right ventricular papillary muscle and rat left ventricular free wall.

Morphological and functional correlates of synchronous beating between embryonic heart cell aggregates and layers

We have examined correlations between morphological and functional evidence of cell coupling between aggregates of beating embryonic heart cells and underlying layers. Synchronously beating aggregate-layer pairs were compared with asynchronous pairs. Intracellular microelectrode studies demonstrated that asynchronously beating aggregate-layers could not be induced to beat synchronously by electrical stimulation of the aggregate, whereas 86% of synchronous instances showed propagation of stimulating current pulses from aggregate to layer. By freeze fracture we have found significant differences both in the number and in the total area of gap junctions between the aggregate-layer interfaces of synchronous and asynchronous preparations. The data suggest that synchronous beating is a reliable functional indication of effective ionic coupling, and requires a certain area and number of gap junction/cell.

Quantitative analysis of low-resistance junctions between cultured cells and correlation with gap-junctional areas.

Electrophysiological studies of low-resistance junctions between Novikoff hepatoma cells grown in suspension cultures were carried out and correlated with gap-junctional areas per inferface determined by freeze-fracture. The mean coupling coefficient between isolated cell pairs was 0.773 +/- 0.025 (SEM) in 67G medium and 0.653 +/- 0.028 in M67 medium; the respective means for the central pairs of four-cell chains were 0.714 +/- 0.034 and 0.595 +/- 0.026. Mean estimates of nonjunctional resistances for cell pairs were 3.0 +/- 0.32 x 10(7) ohm (67G) and 2.01 +/- 0.01 x 10(7) ohm (M67), and the respective estimates for specific nonjunctional resistances were 158.6 +/- 8.1 ohm-cm2 (67G) and 133.0 +/- 812 ohm-cm2 (M67). Mean estimates of junctional conductances were 0.409 +/- 0.058 x 10(-6) mho (67G) and 0.211 +/- 0.018 x 10(-6) mho (M67) for pairs and 0.291 +/- 0.063 x 10(-6) mho (67G) and 0.212 +/- 0.04 mho (M67) for four-cell chains. The mean area of gap junction per interface for separate cell populations was 0.187 +/- 0.049 micron 2 and 0.269 +/- 0.054 micron 2 for cells fixed in loose pellets and in suspension, respectively. When compared with the mean junctional conductance, these values gave specific junctional conductance estimates of 1.13 x 10(2) mho/cm2 and 0.78 x 10(2) mho/cm2, respectively. These values are higher than most previous estimates, but are consistent with the hypothesis that gap-junctional particles contain central hydrophilic channels, about 2 nm in diameter, which have cytoplasmic resistivity.

MORPHOLOGICAL AND FUNCTIONAL CORRELATES OF SYNCHRONOUS BEATING BETWEEN EMBRYONIC HEART CELL AGGREGATES AND LAYERS

In the belief that intercellular communication plays a role in development, we have studied beating 7-day chick embryo heart cell aggregates and layers. In comparison with asynchronous controls we have shown correlations between synchronous beating and both the appearance of gap junctions revealed by freeze-fracture and electro-physiologic evidence of ionic coupling at the interface between them. The freeze-fracture studies reveal gap junctions which are small, often unusual in shape and occur in clusters; these features are also characteristic of the gap junctions in intact hearts from embryos of the same age. When the interfaces between 11 synchronous and 10 asynchronous aggregate-layers were compared there were significant differences in the number of gap junctions and the total area of gap junctions per cell, but no significant differences in the number of cells containing junctions, the percent of total membrane area occupied by junctions, their density or their average size. The electrophysiological studies showed no propagation of stimulating current pulses from aggregate to layer in 16 asynchronous instances, but successful transmission in 6 of 7 synchronous aggregate-layers. These data indicate that synchronous beating is a sensitive detector of the presence of ionic coupling and of gap junctions, suggesting its usefulness in further studies of the role of intercellular communication in development.

Fine structure of the ocellus of the cabbage looper moth (Trichoplusia ni).

The dorsal ocelli of adult cabbage looper moths were studied by light microscopy, and scanning and transmission electron microscopy. Each ocellus has a cuticular lens located on the distal end of a cuticular cone which encapsulates the receptor cells. There are two distinct types of receptor cells in the ocellus. Seventy large receptor cells from plate-like rhabdoms with several adjacent cells to produce a rhabdom network in the ocellus. Proximally ninety small receptor cells which have a disorganized microvillar rhabdomere are located at the base of the rhabdoms formed by the large cells. Extensive areas of gap junctions which occur between the rhabdoms and the membranes of large and small cells suggest that the cells may be electrically coupled to one another. Axons from both large and small receptor cells leave the base of the ocellus and extend to the protocerebrum to synapse with second order neurons.

Hormonal regulation of gap junction differentiation.

Thin-section, tracer, and freeze-cleave experiments on hypophysectomized Rana pipiens larvae reveal that gap junctions form between differentiating ependymoglial cells in response to thyroid hormone. These junctions assemble in large particle-free areas of the plasma membrane known as formation plaques. Between 20 and 40 h after hormone application, formation plaque area increases approximately 26-fold while gap junction area rises about 20-fold. The differentiation of these junctions requires the synthesis of new protein and probably RNA as well. On the basis of inhibitor experiments, it can be reported that formation plaques develop at about 16-20 h after hormone treatment and stages in the construction of gap junctions appear 4-8 h later. These studies suggest that gap junction subunits are synthesized and inserted into formation plaque membrane during the differentiation of the anuran ependymoglial cells.

Intercellular coupling in the atrioventricular node and other tissues of the rabbit heart.

1. A fluorescent tracer dye, sodium fluorescein (mol.wt. 332), was used to assess the relative degree of intercellular coupling in various tissues of the rabbit heart. 2. Dye was injected intracellularly by micro-iontophoresis. Subsequent movement into contiguous cells was monitored by video microscopy. From these data the permeability of the intercellular boundaries was computed. 3. The values of boundary permeability were consistent with those expected from previous studies with tracers whose molecular weights bracketed that of fluorescein. 4. In the atrium, ventricle, Purkinje strands and His bundle, the relative magnitude of the boundary permeability correlated reasonably well with the relative profusity of gap junctional area on the intercalated disk, the latter estimated from published data. 5. The rate of passage of dye between N cells of the atrioventricular, AV, node was at least three orders of magnitude lower than between cells of the other tissues studied; this result is consistent with published reports indicating few gap junctions between cells within the region of slow conduction. 6. Quantitative considerations based on these data indicate that N cells may not be sufficiently well coupled to permit impulse propagation through the AV node by intercellular current flow, alone.

Ultrastructure of the labyrinth in the rat full-term placenta.

The placental labyrinth of the chorioallantoic placenta of the rat was studied with transmission and scanning electron microscopy. Full-term placentas were investigated after perfusion fixation from the maternal and fetal circulation, including thin-sectioning and freeze-fracturing. The labyrinthal three-dimensional structure was found not to be trabecular but spongious lamellar. We propose a division of the lamellae into three groups—the first, second, and third order. The trophoblastic layers are described in detail, showing a cellular layer I and two syncytial layers II and III. Layer I is found to be fenestrated and highly permeable. Layers II and III are connected by extended gap junction areas. It is suggested that the gap junctions function as a molecular sieve and represent the limiting barrier in diaplacental transport of the choriollantoic placenta. Fetal capillaries are fenestrated and endothelial cells are connected by tight junctions. The functional significance of the morphologic findings is discussed.

Morphological correlates of electrotonic coupling in the magnocellular mesencephalic nucleus of the weakly electric fish Gymnotus carapo.

The magnocellular mesencephalic nucleus (MMN) of Gymnotus carapo was studied by electron microscopy. This particular nucleus, characteristic of weakly electric fish, contains two principal classes of neuron. (1) Large neurons (25-35 mum): these are rounded unipolar cells, with the perikaryon partially covered by a sheath of compact myelin. The axon leaves the neuron as a short thick unmyelinated process not resembling the initial segment of multipolar neurons. The axon branches profusely and becomes myelinated very close to its origin. The perikaryal surface not covered by the myelin sheath receives abundant club endings. The synaptic interface between club endings and large neurons is characterized by alternating gap junctions and attachment plaques. In addition, at the periphery of the club endings, "active" zones are generally present, and this synapse is therefore a "mixed" synapse. (2) Small neurons (5-12 mum): these are uni- or bipolar cells, scattered throughout the nucleus, and occasionally, grouped in small clusters. Gap junctions were not observed between neuronal perikarya in such clusters. The synaptic investment of small neurons is formed by long cup endings which almost completely encircle the perikarya. The synaptic interface between cup endings and the perikarya of small neurons is characterized by large areas of gap junctions. A single cup ending establishing gap junctions with two small neurons within the plane of the section was frequently observed and this arrangement provides a morphological basis for electrotonic coupling between small neurons by way of presynaptic fibres. In the neuropil of the MMN, there are abundant synaptic islands constituted by a large axon terminal in synaptic contact with small unidentified profiles; both synaptic elements are surrounded by numerous thin glial lamellae. At the synaptic interface, in the islands, both gap junctions and "active" zones are present. The synaptic islands must also be considered as "mixed" synapses. The morphological results presented here correlate with electrophysiological data (Szabo et al., 1975). The very short delay (0.8-1.3 ms) of the MMS response to the fish's own electric organ discharge can only be explained by the existence of electrotonic transmission along the neuronal chain of the electrosensory pathway. The presence of gap junctions between club endings and large neurons provides a morphological basis for electrotonic transmission at the mesencephalic level of the electrosensory pathway.

Gap junctions in the heart of teleost fish.

An examination of cell contacts was made in the atrial and ventricular muscle of Teleost fish. The intercalated discs consist of two types of junction resembling fascia and macula adherens. Small focal areas of gap junctions were shown to be numerous away from the regions of intercalated disc between the myocardial cells.