Hemicyst formation stimulated by cyclic AMP in dog kidney cell line MDCK

Valentich, J. D.; Tchao, Ruy; Leighton, Joseph

doi:10.1002/jcp.1041000210

Cited by 95 publications

(48 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…T h e tight junction limiting the apical pole is close to the outer surface of the cell layer. Such an inverted orientation of cell polarity has been reported for other epithelial cells cultured under these conditions: L L C P K 1 cells (Wohlwend et al 1985) and M D C K cells (Valentich et al 1979). …”

Section: Culture In Suspensionsupporting

confidence: 74%

Epithelial Cell Polarization In Culture: Orientation of Cell Polarity and Expression of Specific Functions, Studied With Cultured Thyroid Cells

Mauchamp

Chambard

Verrier

et al. 1987

Journal of Cell Science

View full text Add to dashboard Cite

SUMMARYIsolated porcine thyroid cells reorganize in culture into various types of multicellular structure, which differ in the orientation of cell polarity and in the surface of the cell layer accessible to molecules present in the culture medium. T he types of structure are: (1) follicles', the basal pole is oriented toward the medium; (2) inside-out follicles or monolayers: the apical pole is facing the culture medium; (3) monolayers on a perm eable substratum', both sides of the cell layer are accessible to the medium. Follicles can be transformed into inside-out follicles or monolayers and vice versa by manipulation of the external cell environment and without dissociating the cells.Cells concentrate iodide and respond to acute stimulation by thyroid-stimulating hormone (T S H ) when the basal pole is accessible, and organification occurs only when cells form a closed follicular lumen.In porous-bottomed culture chambers monolayers are formed with the basal surface accessible to the medium and the apical compartment separated from the medium. Under these conditions 8 5 -9 5 % of the thvroglobulin produced is secreted apically and 5-15% basally. Thyrotropin stimulates (X3) apical accumulation without modifying secretion in the basal compartment.Sodium transport across the cell layer has been characterized. An amiloride-sensitive influx occurs at the apical pole whereas the Na+/ K +-ATPase, localized in the basolateral membrane, mediates ouabain-sensitive efflux at the basal pole. The thyroid epithelium in culture appears therefore as a Na+-absorbing epithelium. The role of this transport in the stabilization of cell polarity is discussed.

show abstract

Section: Culture In Suspensionsupporting

confidence: 74%

Epithelial Cell Polarization In Culture: Orientation of Cell Polarity and Expression of Specific Functions, Studied With Cultured Thyroid Cells

Mauchamp

Chambard

Verrier

et al. 1987

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…Even though R5 cells formed continuous monolayers, their saturation density was reduced, and dome formation did not occur. Because domes reflect the vectorial transport of solute (Valentich et al, 1979), their absence indicated that R5 cells might not be as polarized as normal MDCK cells. Fluorescent staining of lateral cell borders with the anti-114-kD antibody presented further evidence that the polarity in R5 cells grown on plastic substrata was compromised (unpublished observation).…”

Section: Morphology Of Transformed Mdck Cellsmentioning

confidence: 99%

Multilayering and loss of apical polarity in MDCK cells transformed with viral K-ras.

Schoenenberger

Żuk

Kendall

et al. 1991

The Journal of Cell Biology

View full text Add to dashboard Cite

Abstract. The effects of viral Kirsten ras oncogene expression on the polarized phenotype of MDCK cells were investigated. Stable transformed MDCK cell lines expressing the v-K-ras oncogene were generated via infection with a helper-independent retroviral vector construct. When grown on plastic substrata, transformed cells formed continuous monolayers with epithelial-like morphology. However, on permeable filter supports where normal cells form highly polarized monolayers, transformed MDCK cells detached from the substratum and developed multilayers. Morphological analysis of the multilayers revealed that oncogene expression perturbed the polarized organization of MDCK cells such that the transformed cells lacked an apical-basal axis around which the cytoplasm is normally organized. Evidence for selective disruption of apical membrane polarity was provided by immunolocalization of membrane proteins; a normally apical l l4-kD protein was randomly distributed on the cell surface in the transformed cell line, whereas normally basolateral proteins remained exclusively localized to areas of cell contact and did not appear on the free cell surface. The discrete distribution of the tight junction-associated ZO-1 protein as well as transepithelial resistance and flux measurements suggested that tight junctions were also assembled.These findings indicate that v-K-ras transformation alters cell-substratum and cell-cell interactions in MDCK cells. Furthermore, v-K-ras expression perturbs apical polarization but does not interfere with the development of a basolateral domain, suggesting that apical and basolateral polarity in epithelial ceils may be regulated independently.

show abstract

“…The assay is based on the ability of MadinDarby canine kidney (MDCK) cells to establish specific cell-cell contacts and to generate tissuelike multicellular structures (cysts) composed of polarized epithelial monolayers (10,11,30,52,59,70). We have shown previously that low-level expression of pp60-src in MDCK cells leads to flat, more spread-out cells and disrupts selective cell adhesion components between MDCK cells, but does not have a mitogenic effect (72).…”

mentioning

confidence: 99%

Elevated expression of pp60c-src alters a selective morphogenetic property of epithelial cells in vitro without a mitogenic effect.

Warren¹,

Handel²,

Nelson³

1988

Mol. Cell. Biol.

View full text Add to dashboard Cite

Madin-Darby canine kidney (MDCK) cells are highly differentiated and have retained the morphogenetic properties necessary to form polarized, multicellular epithelial structures (cysts) in vitro that resemble epithelial tissues in vivo. We introduced the c-src gene into MDCK cells to elevate the level of the plasma membrane-associated cellular tyrosine kinase, pp60Or(, to levels two-to ninefold higher than that expressed in parent MDCK cells. Our results revealed a highly discriminatory biological action of pp6C-src on the morphogenetic properties of MDCK cells. Elevated expression of pp60`src conferred on MDCK cells the ability to undergo dramatic changes of cell shape that includes the formation of long cell processes (100 to 200 ,um), never observed in control MDCK cells. The morphogenesis of multicellular epithelial cysts was altered by elevated levels of pp60`csrc and led to predictable distortions of their three-dimensional architecture. However, these cells established morphologically normal cell polarity, formed adhesive epithelial cell-cell contacts indistinguishable from those of control MDCK cells, and exhibited neither focus-forming ability or anchorageindependent growth potential. Finally, we showed that MDCK cells expressing elevated levels of pp60lj'CC exhibit increased phosphorylation of a more limited number of phosphotyrosine-containing proteins than MDCK cells expressing pp6vS'v. We suggest that a natural function of pp60`cu'c is to regulate the morphogenetic properties which determine the shape of differentiated cells and multicellular structures.The cellular tyrosine kinase, pp6O-src, is structurally homologous to the transforming protein of the Rous sarcoma virus, pp6Ov-src, which induces morphological alterations and uncontrolled proliferation of cells. However, the natural biological function of pp6O-src is unknown (4,5,34,35,66,67).The c-src gene has been assayed for biological activity almost exclusively by transformation assays that detect genes which lead to uncontrolled proliferation of cells (9,36,38,41,44,56,57,61). Elevated levels of pp6O-src do not transform chicken or rat fibroblasts in vitro (36, 56), although very high levels of pp60c-src have weak focus-inducing activity in NIH 3T3 cells (38). Thus, pp60f-src can partially mimic the transforming action of pp6Ov-src, but only under exceptional circumstances (34). Since elevated pp60c-src expression in vivo is most abundant in differentiated cells that cannot divide (15,21,45,64,71; also see Discussion), it is possible that pp60-src has a nonmitogenic natural function which does not manifest itself in transformation assays.We have developed a simple in vitro biological assay that detects nonmitogenic, morphoregulatory activities of pp6Osrc kinases (72). The assay is based on the ability of MadinDarby canine kidney (MDCK) cells to establish specific cell-cell contacts and to generate tissuelike multicellular structures (cysts) composed of polarized epithelial monolayers (10,11,30,52,59,70). We have shown previously that low-leve...

show abstract

Hemicyst formation stimulated by cyclic AMP in dog kidney cell line MDCK

Cited by 95 publications

References 52 publications

Epithelial Cell Polarization In Culture: Orientation of Cell Polarity and Expression of Specific Functions, Studied With Cultured Thyroid Cells

Epithelial Cell Polarization In Culture: Orientation of Cell Polarity and Expression of Specific Functions, Studied With Cultured Thyroid Cells

Multilayering and loss of apical polarity in MDCK cells transformed with viral K-ras.

Elevated expression of pp60c-src alters a selective morphogenetic property of epithelial cells in vitro without a mitogenic effect.

Contact Info

Product

Resources

About