Adhesion and Cytosolic Dye Transfer between Macrophages and Intestinal Epithelial Cells

Martin, Carla A.; El-Sabban, Marwan; Zhao, Liming; Burakoff, Robert; Homaidan, Fadia R.

doi:10.3109/15419069809040283

Cited by 25 publications

(25 citation statements)

References 42 publications

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“…All other macrophagic cells previously studied, such as cultured epidermal Langerhans cells (15), peritoneal macrophages (30,31), foam cells of atherosclerotic lesions (32), J774 cells (33), polymorphonuclear cells in a tissue subjected to ischemia-reperfusion (31), and Kupffer cells (34), express Cx43. In addition, gap junctional communication has been identified morphologically and͞or functionally between cultured macro- phages (35)(36)(37)(38)(39). Presumably, all of these cell types were activated to some degree, inasmuch as they were either under culture conditions or present at inflammatory foci, supporting the fact that Cx43 expression is induced upon activation.…”

Section: Discussionmentioning

confidence: 95%

Microglia at brain stab wounds express connexin 43 andin vitroform functional gap junctions after treatment with interferon-γ and tumor necrosis factor-α

Eugenín

Eckardt

Theis

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

208

203

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Gap junctional communication between microglia was investigated at rat brain stab wounds and in primary cultures of rat and mouse cells. Under resting conditions, rat microglia (FITC-isolectin-B4-reactive cells) were sparsely distributed in the neocortex, and most (95%) were not immunoreactive for Cx43, a gap junction protein subunit. At brain stab wounds, microglia progressively accumulated over several days and formed aggregates that frequently showed Cx43 immunoreactivity at interfaces between cells. In primary culture, microglia showed low levels of Cx43 determined by Western blotting, diffuse intracellular Cx43 immunoreactivity, and a low incidence of dye cou- inflammation ͉ macrophage ͉ connexin ͉ cytokines ͉ dye coupling

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Section: Discussionmentioning

confidence: 95%

Microglia at brain stab wounds express connexin 43 andin vitroform functional gap junctions after treatment with interferon-γ and tumor necrosis factor-α

Eugenín

Eckardt

Theis

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

208

203

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“…The presence of functional gap junctions in macrophages has been a highly controversial issue for years (Kane and Bols, 1980;Dean et al, 1988;Polacek et al, 1993;Alves et al, 1996;Porvaznik and MacVittie, 1979;Martin et al, 1998). Reported differences in homocellular and heterocellular coupling involving macrophages and distinct cell types, such as endothelial cells, fibroblasts and epithelial cells, might be due to diverse experimental conditions leading to the expression of soluble and/or membrane-bound molecules capable of modulating cell coupling.…”

Section: Discussionmentioning

confidence: 99%

Modulation of intercellular communication in macrophages: possible interactions between GAP junctions and P2 receptors

Fortes

Pecora

Persechini

et al. 2004

Journal of Cell Science

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Gap junctions are connexin-formed channels that play an important role in intercellular communication in most cell types. In the immune system, specifically in macrophages, the expression of connexins and the establishment of functional gap junctions are still controversial issues. Macrophages express P2X7 receptors that, once activated by the binding of extracellular ATP, lead to the opening of transmembrane pores permeable to molecules of up to 900 Da. There is evidence suggesting an interplay between gap junctions and P2 receptors in different cell systems. Thus, we used ATP-sensitive and -insensitive J774.G8 macrophage cell lines to investigate this interplay. To study junctional communication in J774-macrophage-like cells, we assessed cell-to-cell communication by microinjecting Lucifer Yellow. Confluent cultures of ATP-sensitive J774 cells (ATP-s cells) are coupled, whereas ATP-insensitive J774 cells (ATP-i cells), derived by overexposing J774 cells to extracellular ATP until they do not display the phenomenon of ATP-induced permeabilization, are essentially uncoupled. Western-blot and reverse-transcription polymerase chain reaction assays revealed that ATP-s and ATP-i cells express connexin43 (Cx43), whereas only ATP-s cells express the P2X7 receptor. Accordingly, ATP-i cells did not display any detectable ATP-induced current under whole-cell patch-clamp recordings. Using immunofluorescence microscopy, Cx43 reactivity was found at the cell surface and in regions of cell-cell contact of ATP-s cells, whereas, in ATP-i cells, Cx43 immunoreactivity was only present in cytosolic compartments. Using confocal microscopy, it is shown here that, in ATP-s cells as well as in peritoneal macrophages, Cx43 and P2X7 receptors are co-localized to the membrane of ATP-s cells and peritoneal macrophages.

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“…In the case of cells with obvious distinct cell morphology, the dye transfer assay may be performed using only the permeable dye (calcein-AM or BCECF-AM) (21,26,38). However, if the two cell populations are indistinguishable by size (FSC) and granularity (SSC) parameters, the strategy of labeling one cell population to distinguish donor from recipient cells is necessary (18,23,31,33).…”

Section: Flow Cytometry: a Useful Approach To Study Gap Junctional Comentioning

confidence: 99%

“…This methodology is also a useful tool to compare the influence of cell-type and connexin-type on coupling properties, since changing the donor and recipient cells allows comparison of dye transfer efficiency by different connexin isoforms and distinct cell types (19,21,33,34,39,40). For example, using flow cytometry, Czyz and coworkers (34) observed that Cx43-transfected HeLa cells transferred calcein more efficiently than did Cx40-transfected HeLa cells.…”

Section: Flow Cytometry: a Useful Approach To Study Gap Junctional Comentioning

confidence: 99%

Flow cytometry analysis of gap junction‐mediated cell–cell communication: Advantages and pitfalls

et al. 2006

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Background: Since the first morphological description of the gap junctions use electron microscopy, a considerable number of techniques has been introduced to evaluate gap junction channel functionality, many of which use dye transfer techniques, such as dye injection and fluorescent dye transfer, analyzed by flow cytometry. Methods: To analyze dye transfer, generally one population of cells is incubated with calcein-AM (0.5 lM) for 30 min at 37°C, and the other population was incubated with the lipophilic dye DiIC 18 (3) (10 lM) for 1 h at 37°C; after incubation, these cells were washed five times with PBS and cocultured for different times, and then the dye transfer was analyzed by flow cytometry.

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Adhesion and Cytosolic Dye Transfer between Macrophages and Intestinal Epithelial Cells

Cited by 25 publications

References 42 publications

Microglia at brain stab wounds express connexin 43 andin vitroform functional gap junctions after treatment with interferon-γ and tumor necrosis factor-α

Microglia at brain stab wounds express connexin 43 andin vitroform functional gap junctions after treatment with interferon-γ and tumor necrosis factor-α

Modulation of intercellular communication in macrophages: possible interactions between GAP junctions and P2 receptors

Flow cytometry analysis of gap junction‐mediated cell–cell communication: Advantages and pitfalls

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