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PubMed1992

Development of gap junctions between gastric surface mucous cells during cell maturation in rats.

The development of gap junctions in rat gastric surface mucous cells during their maturation were examined by indirect immunofluorescence and freeze-fracture methods. Monoclonal antibody against liver gap junction protein stained in large spots along the intercellular junctions between mature gastric surface mucous cells. On the other hand, relatively small fluorescent spots were present over the immature surface mucous cells. The freeze-fracture method showed many large irregular gap junctions between mature surface mucous cells. In immature surface mucous cells, the gap junctions were less developed than those of mature cells, and small gap junctions were occasionally associated with tight junctional strands. Quantitatively, the gap junctions in mature cells were larger and more numerous than those in immature cells. These findings show that gap junctions develop during the maturation of surface mucous cells and suggest that the intercellular communication mediated by gap junctions between gastric surface mucous cells plays an important role in the regulation of cell differentiation and in tissue homeostasis.

Animals↗
PubMed1985

Further characterization of the in vitro assay for inhibitors of metabolic cooperation in the Chinese hamster V79 cell line.

12-O-Tetradecanoylphorbol-13-acetate (TPA) has been previously shown to inhibit metabolic cooperation in Chinese hamster V79 cells. An in vitro assay, based on this phenomenon, has been developed to study tumor promoters. Several parameters concerning the metabolic cooperation assay using V79 Chinese hamster cells were further investigated in this report. Pretreatment of the cells with TPA in situ for different periods of time did not result in any detectable change in the inhibition of metabolic cooperation. If cells were replated after TPA treatment, a different result was obtained. There was an apparent decrease in the ability of TPA to inhibit metabolic cooperation when TPA was added back to the TPA-pretreated cultures. However, when TPA was omitted from the TPA pretreated cultures after replating, the inhibition of metabolic cooperation remained high. It was also found that pretreatment of the cells with another chemical, aldrin, exhibited the same pattern as the in situ TPA pretreatment effect on inhibition of metabolic cooperation. In order to obtain a high level of inhibition of metabolic cooperation when using aldrin in this assay, it was determined that the chemical needed to be present for more than one day. Our studies also showed that a 24 h treatment with 6-thioguanine did not kill 6-thioguanine-sensitive cells quickly, nor did it prevent them from performing metabolic cooperation. The relationship of cell density and TPA concentration was also studied. It was observed that a higher cell density required higher TPA concentration to inhibit, maximally, metabolic cooperation. A 'down regulation' type effect was noted when culture was challenged with different concentrations of TPA. These results were interpreted to be consistent with the hypothesis that inhibited gap-junctional intercellular communication is one of the components of tumor promotion.

Aldrin↗
PubMed1994

Feline immunodeficiency virus decreases cell-cell communication and mitochondrial membrane potential.

The in vitro effects of viral replication on mitochondrial membrane potential (MMP) and gap junctional intercellular communication (GJIC) were evaluated as two parameters of potential cellular injury. Two distinct cell types were infected with the Petaluma strain of feline immunodeficiency virus (FIV). Primary astroglia supported acute FIV infection, resulting in syncytia within 3 days of infection, whereas immortalized Crandell feline kidney (CRFK) cells of epithelial origin supported persistent FIV infection in the absence of an obvious cytopathic effect. An examination of cells under conditions that included an infection rate of more than 90% for either population revealed that the astroglia produced about four times more virus than the CRFK cells. The mitochondrial uptake of the cationic fluorescent dye rhodamine 123 in infected astroglia was less than 45% of that of normal control cells, whereas the MMP of the CRFK cells, which produced about one-fourth as much virus, was 80.8% of that of the normal cells. Cell-cell communication between adjacent cells was determined by the recovery of fluorescence following photobleaching of a single cell. In spite of the lower level of innate cell-cell communication among cultured CRFK cells than among astroglia, viral replication resulted in a 30% decrease in the GJIC of both astroglia and CRFK cells. These studies indicate that cell injury, as defined by an inhibition of MMP and GJIC, can occur as a result of persistent and acute infection with the Petaluma strain of FIV.

Animals↗
PubMed1986

Cell-cell interaction and carcinogenesis.

Before carcinogenic stimuli are introduced, a potentially cancerous cell functions normally; growth is controlled and the cell exists in complete harmony with that of surrounding cells in a given tissue. During the process of multi-stage carcinogenesis, such harmony is destroyed and a presumptive initiated cell accomplishes a clonal expansion, neglecting the growth pattern of surrounding normal cells. Recent results from our own laboratory and others strongly suggest that blocked gap-junctional intercellular communication is an important determinant during the process of tumor promotion.

Animals↗
PubMed2000

Cadherin repertoire determines partner-specific gap junctional communication during melanoma progression.

Reduced gap junction activity has long been implicated in tumorigenesis. To elucidate the potential role of intercellular communication in melanoma development, we examined gap junctional capability of melanocytic cells from various stages of tumor progression in coculture models using dye transfer assays. Normal melanocytes coupled with keratinocytes by gap junctional formation, whereas melanoma cells did not. Instead, melanoma cells communicated among themselves and with fibroblasts. This switch in communication partners coincided with a shift from E-cadherin to N-cadherin expression during melanoma development. Forced expression of E-cadherin by adenoviral gene transfer in N-cadherin-expressing melanoma cells restored gap junctional compatibility with keratinocytes. Our data suggest that (1) melanocyte transformation is associated with loss of the pre-existing gap junctional activity with keratinocytes but a concomitant gain of communication with a newly juxtaposed cell type, the fibroblasts, (2) the specificity of gap junctional formation during melanoma development is determined by the cadherin profile on the melanocytic cells and (3) the overall gap junctional activity of melanocytic cells is not reduced with transformation.

Adenoviridae↗
PubMed1992

In vivo growth of C6 glioma cells transfected with connexin43 cDNA.

In order to examine the possible role of intercellular communication via gap junctions in the control of tumor growth, we have transfected C6 glioma cells with connexin43 cDNA. We obtained several clones with variable expression of connexin43. The growth rate of these clones in culture was inversely related to the degree of expression of the transfected cDNA. To examine the growth of these transfected cells in vivo, cells were grown in spinner culture flasks to form spheroids 250-300 microns in diameter. Spheroids of nontransfected C6 cells produced large gliomas. Immunohistochemical and in situ hybridization analyses revealed relatively high levels of connexin43 protein and mRNA in the host tissue, while little of this protein was detected in the glioma. In contrast, spheroids of connexin43-transfected cells grew more slowly and exhibited elevated levels of connexin43 protein and mRNA. These findings suggest that the expression of connexin43 may be associated with the control of brain tumor growth in vivo.

Animals↗
PubMed2005

[Selective heterologous communication among human lung epithelial cells, fibroblasts, and lung cancer PG cells].

OBJECTIVE: To further understand the roles of gap junctional intercellular communication in carcinogenesis and progression of human lung cancer. METHODS: The heterologous communication was characterized among human lung epithelial cells HLEC, human lung fibroblasts HLF, human lung giant carcinoma PG cells and its Cx43 transfectants PG/C4 by using a preloading assay. Cx43 immunofluorescent staining and Northern blot were performed to examine Cx43 gene expression. RESULTS: Although both human lung fibroblasts HLF and epithelial cells HLEC expressed Cx43 and had very strong homologous communication, HLEC cells were unable to communicate with HLF cells. The human lung carcinoma cell line PG was defective of both homologous communication, and heterologous communication with its epithelial origin HLEC. Transfection of Cx43 into PG cells, which rescued PG cells' homologous communication and enhanced heterologous communication with HLF cells, could restore heterologous coupling with HLEC cells. CONCLUSION: Selective heterologous communication exists among different kinds of human lung cells, when human lung carcinoma cells lost coupling with its epithelial origin, Cx43 gene expression is not enough to establish heterologous communication between them.

Cell Communication↗
PubMed1993

[Studies on gap junction formation and junctional communication between cultured endothelial cells and smooth muscle cells].

Gap junction (GJ) between two contacted cells is the structural basis of intercellular communication. Gap junction transfer plays an important role in regulation of cell proliferation. Cultured endothelial cells (EC) and smooth muscle cells (SMC) were injected with fluorescein Lucifer Yellow by iontophoretic injection, and examined under fluorescein microscope. We have demonstrated that there existed homologous and heterologous GJ and contact mediated dye transfer between cultured EC and SMC. LDL and insulin which promote SMC proliferation may inhibit the junctional communication between SMC and EC. Tumor promoter TPA induced almost complete inhibition of junctional communication between EC and SMC. The results indicate that normally regulative signals of metabolism and growth are transferred between EC and SMC via junctional communication to maintain the vessel wall homeostasis. High dosage LDL and insulin may keep SMC away from normal growth control and induce their proliferation by means of inhibiting junctional communication. It may be one of important mechanisms of atherogenesis.

Animals↗
PubMed2003

Genes, neurons and codes: remarks on biological communication.

I examine critically the application of information-theoretic ideas to biological communication during embryonic development and in the functioning central nervous system (CNS). I show that intercellular communication relies mostly on simple signals whose role is to effect a selection among predetermined cellular states. Hence, a crucial role is played by cellular memory, which stabilizes such states. Memory in cells is partly located in the nuclear DNA; no less important however is (phenotypic) memory lying in the cell's organelles and compartments. Because of combinatorial effects in gene expression patterns, cell memory is an enormously powerful mechanism, which also underlies plasticity, and thus constitutes the factor unifying genetic determination, plasticity and learning. Communication in the CNS is analyzed in some detail: here, cellular memory is embodied in anatomy (i.e., cell shape) and neurochemistry. These are the major, relatively "static" factors affecting the routing of neural impulses in the adult CNS. In addition, however, faster channeling is also required: action potentials must be directed to their targets along a few of exponentially many paths, and this dynamic routing is crucial for proper operation. I suggest that collective oscillatory modes may play a role in solving this addressing problem, in the same way that the clock signal gates the operation of man-made computers.

Animals↗
PubMed1997

Filamentous contacts: the ultrastructure and three-dimensional organization of specialized non-synaptic interneuronal appositions in thalamic relay nuclei.

Filamentous contacts are non-synaptic interneuronal junctions characteristic of thalamic relay nuclei. Symmetrical filamentous contacts occur between two dendrites, two somata or a dendrite and a soma; asymmetrical filamentous contacts occur between axon terminals and dendrites, or occasionally somata, chiefly between the large specific afferent axon terminals of the synaptic glomeruli and the shafts of relay cell dendrites. Both are arranged as extensive net-like (reticular) specializations. The strands of the network enclose fenestrae of variable shape and size and, in perpendicular thin sections, appear as stretches of slightly widened intercellular space containing an electron-dense material and bounded by plasma membranes, the cytoplasmic surfaces of which are coated by electron-dense material into which microfilaments appear to insert. The lamina of cytoplasmic material in dendrites and somata is thicker than that in axon terminals and contains distinct electron-dense sub-units. Regular synaptic junctions may be situated like islands within the territory of an asymmetrical filamentous contact, and small spot-like close membrane appositions resembling gap junctions are occasionally seen in the fenestrae adjacent to the strands of both varieties of contact. Bundles of neurofilaments running in different directions, but in a plane parallel to the plasma membrane, are prominent on either side of the symmetrical filamentous contact and on the dendritic side of the asymmetrical variety. The agranular reticulum also exhibits differences between the contact types. Because of their highly specialized ultrastructure and specific distribution, filamentous contacts probably do not serve a purely adhesive function. Their possible role in the establishment and maintenance of orderly connections between cells is discussed but not favoured. Filamentous contacts probably mediate some form of intercellular communication, possibly involving gap junctions.

Intercellular Junctions↗
PubMed2005

Coordinated movement of bile canalicular networks reconstructed by rat small hepatocytes.

Hepatocytes in vivo have a potential for liver regeneration, but it has been very difficult to reconstruct hepatic organoids in vitro. Recent studies have shown that small hepatocytes (SHs) can reconstruct hepatic organoids including functional bile canaliculi (BC). In the present study we analyzed the movement of BC formed in the hepatic organoids, focusing on the coordination of contraction and dilation among cells and the mechanism producing the coordination. Hepatic cells, including SHs, were isolated from an adult rat liver and cultured. Time-lapse images of BC movements were taken and analyzed in cells treated with or without cytochalasin B (CB). Time-lapse images revealed that all BC, regardless of region contracted in a coordinated manner. Actin filaments were observed along the BC even after the BC networks treated with CB dilated markedly. Microinjection of dye was also carried out to investigate the flow thorough BC. Secreted fluorescein from the injected cell flowed along BC, and gap junctional protein connexin 32 was expressed along BC networks, suggesting cell-to-cell communication. Thus, groups of hepatocytes in the hepatic organoids act in a coordinated manner through intercellular communication.

Animals↗
PubMed1999

Cardiac fibrosis and inflammation: interaction with hemodynamic and hormonal factors.

It is generally admitted that the pathogenesis of perivascular and interstitial cardiac fibrosis involves the response to two types of stimuli: a hormonal one, mainly involving the renin-angiotensin-aldosterone system and the more recently described endothelin system, and a hemodynamic stimulus, particularly high blood pressure. We propose in the present review a third step which, although not exclusive, interacts with the hormonal and hemodynamic ones, and involves inflammatory mechanisms. Indeed, hypertension is invariably associated with inflammatory cell infiltration either in the intimal part of large vessels or in the adventitial region of arterioles. This has led us to hypothesize that arterial wall cells may trigger the initial communications attracting inflammatory cells to the perivascular region. In this paper, we review the proinflammatory intercellular communications as well as the intracellular signaling which confer an inflammatory phenotype to arteries. In this context, the profibrogenic and proinflammatory effects of hemodynamic overload and peptidergic systems such as angiotensin II and endothelin are considered. The study of the inflammatory process is not without interest, especially in view of the strong modulating effect of the inflammatory mediators both on the inflammatory process itself and on the fibrotic process. The principal and the most potent mediators are reviewed. Finally, the hypothesis that the inflammatory process could be in reality an immune specific process is suggested.

Angiotensin II↗
PubMed1984

A comparative study on the intercellular canalicular system and intercellular junctions in the pancreatic islets of some rodents.

Intercellular junctions were studied in pancreatic islets of some rodents by electron microscopy of thin sections and freeze-fracture-replicas. Junctions between islet cells can be classified into three types: macula occludens, a combination of the macula occludens and gap junction, and gap junction. The first and second types are mainly located on the cell membrane near the intercellular canaliculus, while the third type is present independently of it. The development of these types of junctions varies considerably among animal species: in mice, guinea pigs and hamsters, the second and third types are frequent, but the first type is rarely seen. In Mongolian gerbils, diminutive elements of the three types are infrequently present. In rats, only the third type is developed. In addition to intercellular communication through gap junctions, the first and second types of junctions may incompletely discriminate the intercellular canalicular lumen, and regulate the microenvironment of the islet cell.

Animals↗
PubMed1986

[Neurochemical aspects of interneuronal communication].

It is suggested that the term neurotransmission, which is used to designate neuronal communication at synaptic level, be associated to the less restrictive term neuromodulation. These two types of intercellular communication seem in fact to be two basically different mechanisms, both of which contribute to neuronal integration. The integration of neuronal information at cellular level appears to be more complex than the simple addition of excitatory plus inhibitory influences eliciting postsynaptic responses. Evidence has been obtained that non synaptic transmission can alter the capacity of a given synapse to transfer neuronal information from the presynaptic element to the postsynaptic neuron. For instance, presynaptic mechanisms provide evidence for the functional independence of the nerve terminals, since the release of neuromediators by the latter is sometimes independent of the axonal firing rate. Similarly, the somato-dendritic part of some neurons exhibits intrinsic functions, such as a dendritic release of neuromediator, suggesting that the control of the axonal firing rate takes place partly at this somato-dendritic level and does not depend for the totality on afferent axonic information. The intercellular operations which organize individual neurons into neuronal networks will also occur either at somato-dendritic level or at the level of specific nerve terminals selected as the result of presynaptic interactions. This integration of neuronal information also seems to take place at postsynaptic level, where cooperative interactions have been shown to occur between various receptors. These mechanisms will function at the level of a single nerve terminal containing more than one neuromediator. Neuromodulation can therefore be said to involve very efficient adaptive processes, which help to account for the fact that such large behavioral responses are expressed by such a small number of neuronal elements.

Animals↗
PubMed2006

Gap junction communications influence upon fibroblast synthesis of Type I collagen and fibronectin.

In rats polyvinyl alcohol sponge subcutaneous implants treated with gap junctional intercellular communications (GJIC) uncouplers showed reduced deposition of connective tissue. Do uncouplers inhibit the synthesis and deposition of a new connective tissue by fibroblasts? Confluent human dermal fibroblasts in serum-free medium received either endosulfan or oleamide, GJIC uncouplers. Collected media were subjected to Dot Blot analysis for native Type I collagen and fibronectin. Uncoupler-treated fibroblasts released less Type I collagen, while there was no change in fibronectin release. Collagen synthesis was restored to normal, when the uncouplers were removed, showing that these uncouplers were reversible and not toxic to cells. Northern blot analysis revealed procollagen alpha1 (I) mRNA was minimally affected by endosulfan. Oleamide-treated 17-day chick embryo calvaria explants were incubated with Type I collagen antibody, frozen, cryosectioned, and then subjected to rhodamine (Rh) tagged anti-mouse-IgG antibody, to detect newly deposited Type I collagen. Fluorescent antibody-collagen complexes were localized on the periphery of cells in control calvaria, but absent around cells in oleamide-treated calvaria. GJIC optimize collagen synthesis but not fibronectin synthesis. The lack of connective tissue deposited in granulation tissues treated with uncouplers appears related to the inhibition of collagen synthesis. These findings suggest that altering GJIC might control collagen deposition in scarring.

Animals↗
PubMed1995

Intercellular signaling in neuronal-glial networks.

Glial cells have recently been found to exhibit electrophysiological and metabolic responses to many neurotransmitters and neuromodulators. These findings have focused attention on the possibility that active signaling between neurons and glia could represent an important form of intercellular communication within the brain. Since glial and neuronal networks are both physically and metabolically interlinked, such intercellular signaling may represent a mechanism for inducing collective changes in the cellular physiology of neuronal and glial cell populations. Within the nervous tissue of both vertebrate and invertebrate organisms, glial cells are known to secrete extracellular signal molecules, modulate carbohydrate metabolism, and control the volume and ionic composition of extracellular space. In this paper, the roles that cytoplasmic [Ca2+] transients may play in regulating these glial cell functions are reviewed. Mechanisms by which intracellular Ca oscillations and intercellular Ca waves may be generated in neurotransmitter-stimulated glial cells are also discussed. In addition, it is proposed that rhythmic glial cell contractions and shape changes, which have been observed for many decades, are linked to Ca-induced secretion of ions, water, and neuroactive compounds. These activities represent mechanisms by which Ca-induced changes in glial cell physiology could potentially alter the excitability of neuronal networks.

Animals↗
PubMed2000

Effects of four inorganic lead compounds on the proliferation and junctional coupling of cultured REL liver cells.

BACKGROUND: Chronic, low-level exposure to inorganic lead (Pb) has been involved in a number of human diseases, including tumors. In this study, the effect of four different inorganic Pb compounds (acetate, chloride, monoxide, and sulfate) was evaluated, in vitro, on liver-derived REL cells, known to be very sensitive to tumor promoters. METHODS: Cytotoxicity and effects on intercellular communication (GJIC) were evaluated, respectively, by cell- density/proliferation and dye-transfer assays. Pb concentration in the media solutions used for each treatment was quantified by atomic absorption spectroscopy-electrothermal atomization. RESULTS: Each of the Pb compounds we tested showed a typical dose- and time-related effect on REL cell proliferation, this effect not being related to the free metal concentration. Contrary to classical tumor promoters, none of the compounds significantly affected REL GJIC (1-hour treatment). CONCLUSIONS: Our results are indicative of specificity in the effects of the different Pb compounds. The mechanism(s) of their action need further investigations.

Animals↗
PubMed1985

cAMP in ovine oocytes: localization of synthesis and its action on protein synthesis, phosphorylation, and meiosis.

In the first series of experiments, the source of cAMP in the sheep oocyte was studied. Cholera toxin was shown to be a potent stimulator of cAMP in isolated sheep oocytes, demonstrating the presence of adenyl cyclase. There was no evidence for transmission of cAMP from stimulated myocardial cell monolayers to cumulus-enclosed oocytes even though the existence of a concentration gradient of cAMP and of intercellular communication were demonstrated. However, gonadotrophin-stimulated follicle shells were able to induce a rise in the cAMP content of denuded or cumulus-enclosed oocytes in the same dish, independently of cell contact. Further experiments were designed to study the effects of a cholera toxin-stimulated rise in cAMP on the maturation of oocytes. When applied to cumulus-oocyte complexes, cholera toxin did not block germinal vesicle breakdown (GVBD), nor the accompanying changes in protein synthesis and phosphorylation, although there was evidence for a delaying effect. There were, however, indications that the toxin was inducing abnormalities that became gross when the concentration was raised to 1 microgram/ml. This high concentration of cholera toxin was able to block the maturation of oocytes in intact, gonadotrophin-treated follicles, although once again abnormalities were evident. We conclude that the role of cAMP in the maturation of the sheep oocyte is different from that proposed in the mouse.

Animals↗