Ion transport in ‘tight’ epithelial monolayers of MDCK cells

1981 ◽  
Vol 59 (2) ◽  
pp. 105-114 ◽  
Author(s):  
N. L. Simmons
Keyword(s):  
Ion Transport ◽  
Mdck Cells ◽  
Epithelial Monolayers

Hyperosmotic and isosmotic shrinkage differentially affect protein phosphorylation and ion transport

2012 ◽  
Vol 90 (2) ◽  
pp. 209-217 ◽  
Author(s):  
Svetlana V. Koltsova ◽  
Olga A. Akimova ◽  
Sergei V. Kotelevtsev ◽  
Ryszard Grygorczyk ◽  
Sergei N. Orlov
Keyword(s):  
Ion Transport ◽  
Protein Phosphorylation ◽  
Cell Volume ◽  
Mdck Cells ◽  
Rat Aorta ◽  
Cellular Functions ◽  
Hypertonic Medium ◽  
Mitogen Activated Protein ◽  
Volume Decrease ◽  
In Cells

In the present work, we compared the outcome of hyperosmotic and isosmotic shrinkage on ion transport and protein phosphorylation in C11-MDCK cells resembling intercalated cells from collecting ducts and in vascular smooth muscle cells (VSMC) from the rat aorta. Hyperosmotic shrinkage was triggered by cell exposure to hypertonic medium, whereas isosmotic shrinkage was evoked by cell transfer from an hypoosmotic to an isosmotic environment. Despite a similar cell volume decrease of 40%–50%, the consequences of hyperosmotic and isosmotic shrinkage on cellular functions were sharply different. In C11-MDCK and VSMC, hyperosmotic shrinkage completely inhibited Na+,K+-ATPase and Na+,Pi cotransport. In contrast, in both types of cells isosmotic shrinkage slightly increased rather than suppressed Na+,K+-ATPase and did not change Na+,Pi cotransport. In C11-MDCK cells, phosphorylation of JNK1/2 and Erk1/2 mitogen-activated protein kinases was augmented in hyperosmotically shrunken cells by ∼7- and 2-fold, respectively, but was not affected in cells subjected to isosmotic shrinkage. These results demonstrate that the data obtained in cells subjected to hyperosmotic shrinkage cannot be considered as sufficient proof implicating cell volume perturbations in the regulation of cellular functions under isosmotic conditions.

Nonmitogenic morphoregulatory action of pp60v-src on multicellular epithelial structures

1987 ◽  
Vol 7 (4) ◽  
pp. 1326-1337
Author(s):  
S L Warren ◽  
W J Nelson
Keyword(s):  
Tyrosine Kinases ◽  
Mdck Cells ◽  
Growth Potential ◽  
Junctional Complex ◽  
Madin Darby Canine Kidney ◽  
Epithelial Monolayers ◽  
Zonula Occludens ◽  
Darby Canine Kidney ◽  
Canine Kidney

Madin-Darby canine kidney (MDCK) cells form polarized, multicellular epithelial structures in vitro. Low-level expression of pp60v-src in MDCK cells elicits plasticity in these multicellular structures. Plasticity was revealed by the displacement of cells from mechanically stressed regions of the epithelial monolayers; however, the two-dimensional relationship between the cells in the remainder of the monolayer was maintained. Electron microscopy of multicellular structures revealed abnormal separation of the lateral membranes of adjacent cells and selective uncoupling of the junctional complex; the zonula adherens was disrupted, but the zonula occludens and desmosomes were retained. Significantly, this result was not accompanied by transformation of the cells, as judged by the absence of anchorage-independent growth potential. These results demonstrate a nonmitogenic biological activity of pp60v-src which is experimentally dissociable from transformation. This morphoregulatory action on higher-order epithelial structures may reflect a function of related cellular tyrosine kinases.

Ion Transport in MDCK Cells

1985 ◽  
pp. 37-50 ◽  
Author(s):  
S. Fernández-Castelo ◽  
J. J. Bolívar ◽  
R. López-Vancell ◽  
G. Beaty ◽  
M. Cereijido
Keyword(s):  
Ion Transport ◽  
Mdck Cells

scholarly journals Nonmitogenic morphoregulatory action of pp60v-src on multicellular epithelial structures.

1987 ◽  
Vol 7 (4) ◽  
pp. 1326-1337 ◽  
Author(s):  
S L Warren ◽  
W J Nelson
Keyword(s):  
Tyrosine Kinases ◽  
Mdck Cells ◽  
Growth Potential ◽  
Junctional Complex ◽  
Madin Darby Canine Kidney ◽  
Epithelial Monolayers ◽  
Zonula Occludens ◽  
Darby Canine Kidney ◽  
Canine Kidney

Madin-Darby canine kidney (MDCK) cells form polarized, multicellular epithelial structures in vitro. Low-level expression of pp60v-src in MDCK cells elicits plasticity in these multicellular structures. Plasticity was revealed by the displacement of cells from mechanically stressed regions of the epithelial monolayers; however, the two-dimensional relationship between the cells in the remainder of the monolayer was maintained. Electron microscopy of multicellular structures revealed abnormal separation of the lateral membranes of adjacent cells and selective uncoupling of the junctional complex; the zonula adherens was disrupted, but the zonula occludens and desmosomes were retained. Significantly, this result was not accompanied by transformation of the cells, as judged by the absence of anchorage-independent growth potential. These results demonstrate a nonmitogenic biological activity of pp60v-src which is experimentally dissociable from transformation. This morphoregulatory action on higher-order epithelial structures may reflect a function of related cellular tyrosine kinases.

Occluding junctions in cultured epithelial monolayers

1981 ◽  
Vol 240 (3) ◽  
pp. C96-C102 ◽  
Author(s):  
M. Cereijido ◽  
I. Meza ◽  
A. Martinez-Palomo
Keyword(s):  
Electrical Resistance ◽  
Mdck Cells ◽  
Freeze Fracture ◽  
Structural Components ◽  
Epithelial Monolayers ◽  
Cation Selectivity ◽  
Occluding Junctions ◽  
Freeze Fracture Electron Microscopy ◽  
Macromolecular Tracers ◽  
Fracture Electron

When MDCK cells are cultured in monolayers, they synthesize, assemble, and seal occluding junctions that limit the paracellular route. These processes may be impaired by inhibitors of the protein synthesis but not by inhibitors of the synthesis of RNA. Once established, the occluding junctions confer to the monolayer an overall electrical resistance of 80–600 omega . cm2. At the microscopical level, the resistance of individual junctions have large variations along the perimeter of a given cell. This agrees with the images of freeze-fracture electron microscopy where the network of the junction varies abruptly from 1 to 10 strands. The junctions are impermeable to macromolecular tracers, have a 9 to 1 Na+/Cl- discrimination, and a cation selectivity following the order: K+ greater than Na+ greater than Rb+ greater than Cs+ greater than Li+. Sealing requires extracellular Ca2+, but the junctions open when the concentration of Ca2+ in the cytoplasm increases. The structural components of the cytoskeleton (microtubules and microfilaments) seem to be involved in the junctional events as revealed by staining with immunofluorescent specific antibodies. If the cells are treated with cytochalasin B, actin microfilaments disorganize, the junctions open, and the electrical resistance across the monolayers falls. The resealing of the tight junction is inhibited by this drug.

scholarly journals Listeria monocytogenesInlP interacts with afadin and facilitates basement membrane crossing

2018 ◽  
Author(s):  
Cristina Faralla ◽  
Effie E. Bastounis ◽  
Fabian E. Ortega ◽  
Samuel H. Light ◽  
Gabrielle Rizzuto ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Mdck Cells ◽  
Cell Junctions ◽  
Host Cells ◽  
Protective Function ◽  
Knock Out ◽  
Placental Infection ◽  
Epithelial Monolayers ◽  
Basal Face ◽  
Cell Cell

ABSTRACTDuring pregnancy, the placenta protects the fetus against the maternal immune response, as well as bacterial and viral pathogens. Bacterial pathogens that have evolved specific mechanisms of breaching this barrier, such asListeria monocytogenes, present a unique opportunity for learning how the placenta carries out its protective function. We previously identified theL. monocytogenesprotein Internalin P (InlP) as a secreted virulence factor critical for placental infection (1). Here, we show that InlP, but not the highly similarL. monocytogenesinternalin Lmo2027, binds to human afadin (encoded byAF-6), a protein associated with cell-cell junctions. A crystal structure of InlP reveals several unique features, including an extended leucine-rich repeat (LRR) domain with a distinctive Ca2+-binding site. Despite afadin’s involvement in the formation of cell-cell junctions, MDCK epithelial cells expressing InlP displayed a decrease in the magnitude of the traction stresses they could exert on deformable substrates, similar to the decrease in traction exhibited byAF-6knock-out MDCK cells.L. monocytogenes ΔinlPmutants were deficient in their ability to form actin-rich protrusions from the basal face of polarized epithelial monolayers, a necessary step in the crossing of such monolayers (transcytosis). A similar phenotype was observed for bacteria expressing an internal in-frame deletion ininlP(inlPDLRR5) that specifically disrupts its interaction with afadin. However, afadin deletion in the host cells did not rescue the transcytosis defect. We conclude that secreted InlP targets cytosolic afadin to specifically promoteL. monocytogenestranscytosis across the basal face of epithelial monolayers, which may contribute to the crossing of the basement membrane during placental infection.

scholarly journals Control and mechanisms of pulsatile flows in epithelial monolayers

2020 ◽  
Author(s):  
Raghavan Thiagarajan ◽  
Alka Bhat ◽  
Guillaume Salbreux ◽  
Mandar M. Inamdar ◽  
Daniel Riveline
Keyword(s):  
Mdck Cells ◽  
Cell Polarization ◽  
Cellular Mechanisms ◽  
Epithelial Monolayers ◽  
Local Contraction ◽  
Collective Movements ◽  
Pulsatile Flows ◽  
Contraction And Expansion

Epithelial cells flows are observed both in vivo and in vitro and are essential for morphogenesis. Here, we show that pulsatile flows involving local contraction and expansion of a tissue can arise in vitro in an epithelial monolayer of Madine Darby Canine Kidney (MDCK) cells. The strength of pulsation can be modulated through friction heterogeneity by observing the monolayer dynamics on micro-contact printed fibronectin grids with dimensions matching the length-scale of spontaneous oscillations. We also report pulsations by inducing wound closure in domains of similar size with micro-fabricated pillars. In contrast, strongly coherent flows can be induced by adding and washing out acto-myosin cytoskeleton inhibitors. To gain insight into the associated cellular mechanisms, we fluorescently label actin and myosin. We find that lamellipodia align with the direction of the flow, and tissue-scale myosin gradients arise during pulsations in wound-healing experiments. Pulsations and flows are recapitulated in silico by a vertex model with cell motility and polarisation dynamics. The nature of collective movements depends on the interplay between velocity alignment and random diffusion of cell polarisation. When they are comparable, a significant pulsatile flow emerges, whereas the tissue undergoes long-range flows when alignment dominates. We conjecture that the interplay between lamellipodial motile activity and cell polarization, with a possible additional role for tissue-scale myosin gradients, is at the origin of the pulsatile nature of the collective flow. Altogether, our study reveals that monolayer dynamics is dictated by simple rules of interaction at cellular levels which could be involved in morphogenesis.

scholarly journals Pili Binding to Asialo-GM1 on Epithelial Cells Can Mediate Cytotoxicity or Bacterial Internalization byPseudomonas aeruginosa

1999 ◽  
Vol 67 (7) ◽  
pp. 3207-3214 ◽  
Author(s):  
James C. Comolli ◽  
Leslie L. Waite ◽  
Keith E. Mostov ◽  
Joanne N. Engel
Keyword(s):  
Epithelial Cells ◽  
Cell Injury ◽  
Mdck Cells ◽  
Type Iv Pili ◽  
Receptor Interaction ◽  
Apical Surface ◽  
Epithelial Monolayers ◽  
Type Iv ◽  
Mediate Cytotoxicity ◽  
Asialo Gm1

ABSTRACT The interaction of Pseudomonas aeruginosa type IV pili and the glycosphingolipid asialo-GM1 (aGM1) can mediate bacterial adherence to epithelial cells, but the steps subsequent to this adherence have not been elucidated. To investigate the result of the interaction of pili and aGM1, we used polarized epithelial monolayers of Madin-Darby canine kidney (MDCK) cells in culture, which contained little detectable aGM1 on their apical surface but were able to incorporate exogenous aGM1. Compared to an untreated monolayer,P. aeruginosa PA103 displayed an eightfold increase in association with and fivefold more cytotoxicity toward MDCK cells pretreated with aGM1. Cytotoxicity of either carrier-treated or aGM1-treated monolayers required the type III secreted protein ExoU. Asialo-GM1 pretreatment of MDCK monolayers likewise augmented bacterial internalization of an isogenic invasive strain approximately fourfold. These increases were not seen in monolayers treated with GM1, the sialyated form of the glycolipid, and were inhibited by treatment with an antibody to aGM1. Also, the aGM1-mediated adhesion, cytotoxicity, and internalization required intact type IV pili since nonpiliated PA103 mutants were unaffected by aGM1 pretreatment of MDCK cells. These results demonstrate that epithelial cell injury and bacterial internalization can proceed from the same adhesin-receptor interaction, and they indicate that P. aeruginosa exoproducts solely determine the steps subsequent to adhesion.

Transfection Alters Ion Transport in MDCK Cells

1996 ◽  
Vol 149 (1) ◽  
pp. 49-55 ◽  
Author(s):  
D.M. Kaji *, † , J. Bates ◽  
J.D. Goyzueta *, † , K. Prasadan ◽  
H. Yu ◽  
S. Kumar
Keyword(s):  
Ion Transport ◽  
Mdck Cells
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