Dynamics of E-cadherin and catenin complexes γ-catenin complexes during dedifferentiation of polarized MDCK cells

Balkovetz, Daniel F.; Sambandam, Vijaya

doi:10.1046/j.1523-1755.1999.00623.x

Cited by 24 publications

(25 citation statements)

References 40 publications

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“…Previous studies have analyzed the effects of HGF on intestinal or renal epithelial permeability to inulin and have obtained results similar to ours showing a slight increase in inulin flux compared with untreated monolayers. However, the authors of these previous studies have drawn contrasting conclusions, suggesting either that marginal increases in inulin permeability induced by HGF indicate that monolayer integrity is preserved or that small increases in inulin flux indicate a loss of cell-cell junctional integrity (5,53). In our study, in addition to testing the inulin permeability of control and HGF-treated MDCK cell monolayers, we also analyzed inulin permeability across monolayers that lack TJs by testing confluent MDCK cell monolayers in which cell-cell adhesion was removed by treatment with low-Ca 2ϩ medium.…”

Section: Discussionmentioning

confidence: 98%

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Hepatocyte growth factor induces MDCK cell morphogenesis without causing loss of tight junction functional integrity

Pollack¹,

Apodaca²,

Mostov³

2004

American Journal of Physiology-Cell Physiology

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Hepatocyte growth factor (HGF) induces mitogenesis, motogenesis, and tubulogenesis of cultured Madin-Darby canine kidney (MDCK) epithelial cells. We report that in addition to these effects HGF stimulates morphogenesis of tight, polarized MDCK cell monolayers into pseudostratified layers without loss of tight junction (TJ) functional integrity. We tested TJ functional integrity during formation of pseudostratified layers. In response to HGF, the TJ marker ZO-1 remained in morphologically complete rings and functional barriers to paracellular diffusion of ruthenium red were maintained in pseudostratified layers. Transepithelial resistance (TER) increased transiently two-to threefold during the morphogenetic transition from monolayers to pseudostratified layers and then declined to baseline levels once pseudostratified layers were formed. In MDCK cells expressing the trk/met chimera, both HGF and NGF at concentrations of 2.5 ng/ml induced scattering. However, 2.5 ng/ml HGF did not affect TER. The peak effect of HGF on TER was at a concentration of 100 ng/ml. In contrast, NGF at concentrations as high as 25 g/ml had no effect on TER or pseudostratified layer morphogenesis of trk/met-expressing cultures. These results suggest that altered presentation of the stimulus, such as through HGF interaction with low-affinity sites, may change the downstream signaling response. In addition, our results demonstrate that HGF stimulates pseudostratified layer morphogenesis while inducing an increase in TER and maintaining the overall tightness of the epithelial layer. Stimulation of epithelial cell movements by HGF without loss of functional TJs may be important for maintaining epithelial integrity during morphogenetic events such as formation of pseudostratified epithelia, organ regeneration, and tissue repair. c-met protooncogene; transepithelial resistance; Madin-Darby canine kidney cell MORPHOGENETIC CELL REARRANGEMENTS are stimulated by hepatocyte growth factor (HGF), a fibroblast-derived growth factor (77) that has paracrine actions on a variety of cell types. In vivo, HGF is important for the development of the placenta, liver, limbs, and kidney (6,28,67,71,72,82,87) and induces morphogenesis of blood vessels (9,22,47,70). In addition, both HGF and the c-met protooncogene (c-met) (31, 86), a tyrosine kinase receptor (11,19,51,54) to which HGF binds with high affinity (25,32), are regulated in response to organ injury and stimulate epithelial morphogenesis that contributes to tissue repair and regeneration in liver, kidney, lung, gastric mucosa, and muscle (12,38,40,66,81,88). In vitro, many of the pleiotropic effects of HGF can be modeled and show a dependence on both the target cell type and environmental context: paracrine actions of HGF increase hepatocyte mitogenesis (20,41,49,50,90), inhibit growth of tumor cells (69, 78), activate motogenesis and invasiveness of epithelial and endothelial cells (9,18,63,64,70,75,84,85), stimulate wound repair (53), and induce morphogenesis of either epithelial tubes or pseudostrati...

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Section: Discussionmentioning

confidence: 98%

“…Transwell filter samples were imaged either in the X-Y plane or in the X-Z plane with a motor step size of 0. 5 …”

Section: Methodsmentioning

confidence: 99%

Hepatocyte growth factor induces MDCK cell morphogenesis without causing loss of tight junction functional integrity

Pollack¹,

Apodaca²,

Mostov³

2004

American Journal of Physiology-Cell Physiology

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“…MDCK Culture, HGF/4-HT Treatment, and RNA IsolationLow passage type II MDCK cells were obtained from K. Mostov (University of California, San Francisco) and used between passages 3 and 10 as described previously (23,24). These cells were originally cloned by Daniel Louvard at the European Molecular Biology Laboratory (EMBL) and came to Keith Mostov via Karl Matlin.…”

Section: Methodsmentioning

confidence: 99%

Matrix Metalloproteinase 13 (MMP13) and Tissue Inhibitor of Matrix Metalloproteinase 1 (TIMP1), Regulated by the MAPK Pathway, Are Both Necessary for Madin-Darby Canine Kidney Tubulogenesis

Hellman

Spector

Robinson

et al. 2008

Journal of Biological Chemistry

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A classic model of tubulogenesis utilizes Madin-Darby canine kidney (MDCK) cells. MDCK cells form monoclonal cysts inthree-dimensional collagen and tubulate in response to hepatocyte growth factor, which activates multiple signaling pathways, including the mitogen-activated protein kinase (MAPK) pathway. It was shown previously that MAPK activation is necessary and sufficient to induce the first stage of tubulogenesis, the partial epithelial to mesenchymal transition (p-EMT), whereas matrix metalloproteinases (MMPs) are necessary for the second redifferentiation stage. To identify specific MMP genes, their regulators, tissue inhibitors of matrix metalloproteinases (TIMPs), and the molecular pathways by which they are activated, we used two distinct MAPK inhibitors and a technique we have termed subtraction pathway microarray analysis. Of the 19 MMPs and 3 TIMPs present on the Canine Genome 2.0 Array, MMP13 and TIMP1 were up-regulated 198-and 169-fold, respectively, via the MAPK pathway. This was confirmed by two-dimensional and three-dimensional real time PCR, as well as in MDCK cells inducible for the MAPK gene Raf. Knockdown of MMP13 using short hairpin RNA prevented progression past the initial phase of p-EMT. Knockdown of TIMP1 prevented normal cystogenesis, although the initial phase of p-EMT did occasionally occur. The MMP13 knockdown phenotype is likely because of decreased collagenase activity, whereas the TIMP1 knockdown phenotype appears due to increased apoptosis. These data suggest a model, which may also be important for development of other branched organs, whereby the MAPK pathway controls both MDCK p-EMT and redifferentiation, in part by activating MMP13 and TIMP1.

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“…43,44 Seventy-two hours after transfection, COS-7 cells were incubated for 20 minutes at 37°C in MEM (MEM Select-Amine Kit, GibcoBRL), which did not contain cysteine, methionine, or serum. Cells were pulse-labeled for 30 35 S-label, ICN Biochemicals), 300 Ci/mL.…”

Section: Metabolic Labeling Study To Determine Nos2 Half-lifementioning

confidence: 99%

Nitric Oxide Synthase (NOS2) Mutation in Dahl/Rapp Rats Decreases Enzyme Stability

Ying

Xia

Sanders

2001

Circulation Research

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Abstract-The pathogenesis of salt-sensitive hypertension remains poorly defined, but a role for nitric oxide (NO) has been suggested. The Dahl/Rapp salt-sensitive rat possesses a defect in NO synthesis that is overcome by supplementation with L-arginine, which increases NO and cGMP production and prevents salt-sensitive hypertension. An S714P mutation of inducible NO synthase (NOS2) was subsequently identified. The current report examined the functional significance of an S714P mutation in NOS2. COS-7 cells were transiently transfected with cDNA of wild-type NOS2 and S714P and S714A mutants of NOS2, and enzyme function was determined. Whereas steady-state mRNA levels did not differ, immunoblot analysis demonstrated decreased levels of NOS2 protein.Metabolic labeling experiments confirmed a reduced half-life of the S714P mutation. Nitrite production, which was dependent on the concentration of L-arginine in the medium, was diminished in cells transfected with the S714P mutant, compared with the wild type and the S714A mutant. These data provide a biochemical explanation of the physiological abnormalities of NOS2 in the Dahl/Rapp salt-sensitive rat and suggest that a posttranslational mechanism involving the proteasome may be responsible for the diminished NO production observed in response to increased dietary salt intake in these animals. Key Words: hypertension Ⅲ nitric oxide Ⅲ nitric oxide synthase Ⅲ proteasome Ⅲ lactacystin A lmost a quarter of the population of the United States has hypertension. 1 Although African-Americans are much more likely to exhibit salt sensitivity, 50% to 75% of the total hypertensive population demonstrate salt sensitivity; that is, blood pressure is altered as dietary salt intake is changed. 2 Breeding studies in rats have confirmed that there is a genetic susceptibility to develop salt-sensitive hypertension. 3,4 Whereas multiple genes are involved in the blood pressure response to changes in dietary salt intake, increasing evidence supports an important role of altered synthesis of nitric oxide (NO) in salt sensitivity. NO synthesis occurs in the vessel wall 5-7 and in the kidney 8 -10 and serves as a locally produced vasodilator 5-7,11 and natriuretic factor. 12-16 Kidney production of NO appears to be particularly important in blood pressure regulation, because infusion of N G -nitro-Larginine methyl ester directly into the renal medulla causes sodium retention and increased blood pressure. 17 Thus, NO is uniquely situated to control blood pressure responses to increased salt intake.Dietary salt modulates NO production in healthy humans 18 and in normotensive rats. 19,20 An increase in salt intake increases NO production. 19,20 Along with determining the blood pressure response to a high-salt diet, 12,19,21,22 the renal vascular resistance and renal functional changes that occur with an increase in dietary salt are also modulated by NO. 12,13,23 In contrast to normotensive rats, the increase in NO production is absent in Dahl/Rapp salt-sensitive (S) rats, an inbred strain of r...

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Dynamics of E-cadherin and catenin complexes γ-catenin complexes during dedifferentiation of polarized MDCK cells

Cited by 24 publications

References 40 publications

Hepatocyte growth factor induces MDCK cell morphogenesis without causing loss of tight junction functional integrity

Hepatocyte growth factor induces MDCK cell morphogenesis without causing loss of tight junction functional integrity

Matrix Metalloproteinase 13 (MMP13) and Tissue Inhibitor of Matrix Metalloproteinase 1 (TIMP1), Regulated by the MAPK Pathway, Are Both Necessary for Madin-Darby Canine Kidney Tubulogenesis

Nitric Oxide Synthase (NOS2) Mutation in Dahl/Rapp Rats Decreases Enzyme Stability

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