Gap junctions in hemodichorial and hemotrichorial placentae

Gap junctions were found to be a constant feature of chorioallantoic placentae with two or three trophoblastic layers. The gap junctions connect layers I and II in hemodichorial and layers II and III in hemotrichorial placentae. Although the gap junctions vary in form and in the packing density of membrane-associated particles, they cover an extensive surface area in all species examined. The gap junctions always connect adjacent membranes of two trophoblastic layers, which show no evidence of micropinocytotic activity; at least one of these trophoblastic layers is syncytial. It is therefore concluded that the gap junctions play an important role in diaplacental transport. We consider that gap junctions act as molecular sieves, resulting in limitations in the transport of large molecules. The passage of small molecules, on the contrary, would be facilitated by the gap junctions.     

Gap junctions in blood forming tissues

More than ten research groups have now reported the presence of gap junctions in blood-forming tissue or cultured cells. It is time to accept that these cell-coupling structures are present in this tissue. To find out what they are doing here we need to develop appropriate experimental techniques. This review covers the particular problems of investigating direct cell-cell communication by gap or other junctions in undisturbed haemopoietic tissue. It then describes and assesses the published reports of haemopoietic gap junctions. Recently, in the author's laboratory, three means of increasing the number of gap junctions 50- to 100-fold in mouse marrow have been described, as well as techniques for doing so in culture. There is a complete report of this work here. At present it is quite unclear what function gap junctions serve in blood-formation, perhaps it is some consolation that 30 years after their ultramicroscopic discovery it is also true for all other unexcitable tissues. Possibly the ability to up-regulate their expression in haemopoietic tissue will help us find out what their role is in blood formation.     

Gap junctions: getting the message through

Connexin proteins make intercellular channels - gap junctions - which provide a direct pathway for cell-cell signaling in vertebrates. Studies of mice lacking connexin genes have demonstrated the need for intercellular transfer of messenger molecules and are uncovering the specific functions of each connexin.     

Gap junctions mediate intercellular calcium signalling in cultured articular chondrocytes

Gap junction-mediated intercellular communication has been implicated in a variety of cellular functions. Among these, signal transduction can be coordinated among several cells due to gap junctional permeability to intracellular second messengers. Chondrocytes from articular cartilage in primary culture respond to extracellular ATP by rhythmically increasing their cytosolic Ca2+ concentration. Digital imaging fluorescence microscopy of Fura-2 loaded cells was used to monitor Ca2+ in confluent and semi-confluent cell layers. Under these conditions, Ca2+ spikes propagate from cell to cell giving rise to intercellular Ca2+ waves. The functional expression of gap junctions was assessed, in confluent chondrocyte cultures, by the intercellular transfer of Lucifer yellow dye in scrape-loading experiments. Intercellular dye transfer was blocked by the gap junction inhibitor 18 alpha-glycyrrhetinic acid. In imaging experiments, the inhibitor caused the loss of synchrony of ATP-induced Ca2+ oscillations, and blocked the intercellular Ca2+ propagation induced by mechanical stimulation of a single cell in a monolayer. It is concluded that gap junctions mediate intercellular signal transduction in cartilage cells and may provide a mechanism for co-ordinating their metabolic activity.     

Gap junctions and growth control in liver regeneration and in isolated rat hepatocytes

The hepatocytes in the mature normal liver are tightly coupled through gap junctions, except during compensatory hyperplasia (regeneration) after partial hepatectomy when the gap junctions become down-regulated. The significance of this down-regulation has been a long-standing enigma. The present study of hepatocytes in primary culture and in the regenerating liver aimed at defining the relationship, if any, between hepatocyte gap junctional communication and proliferation. Gap junctional down-regulation in the regenerating liver appeared to be a specific phenomenon because desmosomes and the surface contact area between neighboring hepatocytes remained constant. All agents and conditions (dexamethasone in vivo; dexamethasone, cyclic adenosine monophosphate, serum, and high cell density in vitro) delaying gap junctional down-regulation also increased the lag before the cells reached competence to enter S phase. This raised the possibility that hepatocyte DNA replication was inhibited through preservation of gap junctions. However, we disproved this assumption by showing that the DNA replication (more specifically the G1/S transition rate constant) was inhibited even in hepatocytes completely devoid of gap junctional communication. The teleological advantage of linking gap junctional down-regulation to hepatocyte G1 progression therefore may not be to trigger DNA replication but to ensure that proliferating hepatocytes and hepatocytes responsible for liver-specific metabolic functions maintain separate pools of metabolites and signaling molecules.     

Gap junctions with amacrine cells provide a feedback pathway for ganglion cells within the retina

In primates, one type of retinal ganglion cell, the parasol cell, makes gap junctions with amacrine cells, the inhibitory, local circuit neurons. To study the effects of these gap junctions, we developed a linear, mathematical model of the retinal circuitry providing input to parasol cells. Electrophysiological studies have indicated that gap junctions do not enlarge the receptive field centres of parasol cells, but our results suggest that they make other contributions to their light responses. According to our model, the coupled amacrine cells enhance the responses of parasol cells to luminance contrast by disinhibition. We also show how a mixed chemical and electrical synapse between two sets of amacrine cells presynaptic to the parasol cells might make the responses of parasol cells more transient and, therefore, more sensitive to motion. Finally, we show how coupling via amacrine cells can synchronize the firing of parasol cells. An action potential in a model parasol cell can excite neighbouring parasol cells, but only when the coupled amacrine cells also fire action potentials. Passive conduction was ineffective due to low-pass temporal filtering. Inhibition from the axons of the coupled amacrine cells also produced oscillations that might synchronize the firing of more distant ganglion cells.     

Gap junctions and nerve terminals among stromal cells in human fallopian tube ampullary mucosa

The purpose of the present study is to investigate the ultrastructure and immunohistochemistry of the stromal cells and terminal nerve fibers in human fallopian tube ampullar mucosa to achieve a detailed characterization of this tissue to permit a better assessment of possible functions. Tissues were obtained during surgery or at autopsy from 26 patients. Specimens were studied by the conventional histologic technique, immunohistochemistry (Cx43, synaptophysin, neurofilament proteins, and S-100 protein), and electron microscopy. Gap junction and nerve terminal frequency between stromal cells were studied by direct assessment on ultrathin sections in the transmission electron microscope. Gap junctions were observed between the cytoplasmic processes of subepithelial stromal cells. There were approximately 23 gap junctions per 73 nucleated stromal spindle cells. Immunohistochemistry using Cx43 antibody confirmed the dot-like distribution of gap junctions. The frequent and intimate association of stromal cell processes with nerve terminals was also demonstrated. Nerve terminals were immunostained by antibodies to nerve-specific molecules and ultrastructurally as axonal profiles containing dense-cored granules or empty vesicles. Analysis of nerve terminal frequency revealed 18 nerve profiles containing 51 axonal profiles per 73 nucleated stromal spindle cells. The present paper documents the participation of autonomic nerve endings and gap junctions in the stromal cell network in human fallopian tube stroma. Similarities to the unique anatomical unit referred to as the 'neuro-reticular complex' in bone marrow tissue (Yamazaki and Allen, 1990) are discussed.     

Mineralization kinetics of air bubbles allowing for the particle detachment and time of buoying of aggregates

The joint consideration of subprocesses of particle capture and detachment, and buoying of aggregates in the periodic nonfrothing flotation conditions shows that the mineral load formed on a separate part for its buoying time (τ_m). This load is a part of the equilibrium mineral load, which can be attained under the infinite mineralization time. It is proposed to characterize the composition and attainment rate of the mineral load by two dimensionless parameters, which depend on intensities of subprocesses. The sort parameter of particles (B) has been uniquely determined by the ratio of the detachment intensity to the capture intensity, while the dimensionless time (D) is determined by the ratio of the particle capture and detachment rate to the buoying velocity of the air bubble. The mineralization kinetic equation by many bubbles is derived in the exponential form similarly to the first-order Beloglazov equation. Intensities of capture and detachment subprocesses in the mineralization rate constant (K_m) determine the magnitude of recovery by a separate bubble (ε_bm) for time τ_m, while the air consumption determines the summary recovery ε.     

Influence of trematode parasitism on the growth of a bivalve host in the field

Trematode-induced gigantism of molluscs, enhanced growth of trematode-parasitised individuals, has been observed in many laboratory studies. This study reports the effect of the sterilising trematode, Rhipidicotyle fennica, on the growth of the freshwater clam Anodonta piscinalis under field conditions. In addition to single infections (prevalence 44%), a few clams (3%) were infected with both R. fennica and Rhipidicotyle campanula. Parasite-induced gigantism was not found; parasites lowered host growth. The decreased in growth was correlated with the quantity of parasite material. Clams with double infections grew the least, although they did not differ significantly from hosts with a heavy single species infection. The growth of the experimental clams was lower than that of undisturbed control clams. Trematode-induced gigantism of molluscs in the field, in general, remains undemonstrated.     

Electrical properties of the rod syncytium in the retina of the turtle

1. Intracellular responses were recorded from rods in isolated eye-cups of the snapping turtle. Chelydra serpentina. Responses to flashes of small (less than 100 mum diameter) and large (1000 mum diameter) spots of 500 nm light were studied. 2. Responses produced by small and large diameter spots which delivered less than 0-3 photons mum-2 had the same shape. The responses produced by large spots were, however, nearly ten times greater in amplitude. The difference in amplitude is termed enhancement. 3. Perfusing an eye-cup with a Co2+-containing medium blocked synaptic transmission from receptors to horizontal cells but did not affect the responses of rods. 4. The membrane conductance of a single rod, estimated by three independent methods, was approximately 1-2 X 10(-9) MHo. 5. Enhancement can be predicted by a mathematical model which treats rods as an electrical syncytium. The space coefficient describing the spread of current is approximately 65 mum indicating that the coupling conductance between rods was relatively high. 6. When the intensity of a small spot was increased from 0-3 photons mum-2 up to 6 photons mum-2, the shape of the response was unchanged. When the intensity of a large spot was increased to more than 0-3 photons mum-2, the voltage during the recovery phase was decreased. This decrease is termed disenhancement. 7. The voltages produced by bright, large and small diameter spots which delivered the same quantity of light to the impaled rod were compared. The voltage produced by a large diameter spot became for a short period during the recovery phase less than the voltage produced by a small diameter spot. This observation indicates that the response to a large spot included during recovery an active process which is not apparent in the response to a small spot.     

Evidence for a major gene controlling susceptibility to tegumentary leishmaniasis in a recently exposed Bolivian population

Tegumentary leishmaniasis due to Leishmania braziliensis is a parasitic disease that occurs in two stages after the infected sandfly bite: (1) a primary cutaneous lesion followed by (2) a secondary mucosal involvement generally resulting in severe facial deformities. In order to investigate the genetic and environmental factors involved in the development of the cutaneous lesion, a familial study was performed in a region of Bolivia in which the disease is endemic. Complete selection of 118 nuclear families (703 subjects, with 241 patients), each with at least one cutaneous affected subject, was achieved; 41 families were of native origin, and 77 (herein designated "migrant") recently had settled in the area. For the analysis, the trait under study was the time to onset of the primary cutaneous lesion. The start of the follow-up was birth, for native population, or date of arrival in the endemic area, for migrant population. Segregation analysis was performed by use of a model based on survival analysis methods that allows joint estimation of genetic and environmental effects and accounts for gene x covariate interactions. A significant effect of gender, home-forest distance, and forest-related activity was found. In the 77 migrant families there was evidence for a recessive major gene controlling the onset of the primary cutaneous lesion, with residual familial dependences and age x genotype interaction. Penetrance estimations show that young subjects are genetically more susceptible than older subjects, suggesting that this genetic component could concern mechanisms involved in the development of individual protection during childhood. There was also a significant genetic heterogeneity of the sample according to the native/migrant origin of the families, and no major-gene effect was found in the native subsample.     

Nonlocal interaction in a system of Josephson junctions

Trisomy 8 is a frequently acquired cytogenetic abnormality in myeloid malignancies, but may also represent a constitutional chromosome abnormality with a wide phenotypic variation. We report a case of myelodysplastic syndrome (MDS) that developed in a child with trisomy 8 mosaicism and normal phenotype. Bone marrow (BM) cells all showed trisomy 8 with additional clonal abnormalities in most cells. Based on the present case and a review of previously published cases of myeloid malignancies in patients with trisomy 8 mosaicism, it appears likely that the malignant cells developed from the trisomic cell population, suggesting that constitutional trisomy 8 may be a predisposing condition to myeloid malignancies. Trisomy 8 in malignant cells is usually considered an acquired abnormality, but this implies a risk of ignoring a constitutional trisomy 8 mosaicism. Examination for constitutional trisomy 8, despite a normal phenotype, may therefore be warranted in hematologic malignancies with trisomy 8 of BM cells to evaluate further the possible association and to preclude erroneous use of trisomy 8 as a tumor marker.     

Recombinant antigens for specific and sensitive serodiagnosis of Latin American tegumentary leishmaniasis

The diagnostic potential of recombinant leishmanial antigens for Latin American tegumentary leishmaniasis (LATL) was examined. Two Leishmania (Viannia) peruviana recombinant proteins, T26-U2 and T26-U4, were assessed by their reactivity to detect specific anti-leishmanial antibodies. Seventy-eight individual sera from persons with LATL, 39 from those with other diseases, and 10 negative control sera were tested by Western blotting and enzyme-linked immunosorbent assay. The sensitivity of the test using T26-U2 plus T26-U4 was similar to that obtained with whole parasite extract (92%). However, the specificity obtained using both recombinant antigens (87%) was higher than that of the whole parasite extract (65%). All tests using recombinant proteins (T26-U2, T26-U2 plus T26-U4 or T26-U4) had a higher positive predictive value (89%, 92% and 98%, respectively) than the value obtained using total parasites (81%). Eleven Colombian sera were also tested, and the results indicated that T26-U2 plus T26-U4 could be used successfully in Peru and in other Latin American countries.