scholarly journals HGF-induced serine 897 phosphorylation of EphA2 regulates epithelial morphogenesis of MDCK cells in 3D culture

2015 ◽  
Vol 128 (10) ◽  
pp. 1912-1921 ◽  
Author(s):  
K. Harada ◽  
M. Negishi ◽  
H. Katoh
Keyword(s):  
Mdck Cells ◽  
3D Culture ◽  
Epithelial Morphogenesis

scholarly journals Epithelial morphogenesis of MDCK cells in three-dimensional collagen culture is modulated by interleukin-8

2013 ◽  
Vol 304 (10) ◽  
pp. C966-C975 ◽  
Author(s):  
Erika K. Wells ◽  
OrLando Yarborough ◽  
Richard P. Lifton ◽  
Lloyd G. Cantley ◽  
Michael J. Caplan
Keyword(s):  
Gene Expression ◽  
Mdck Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Collagen Gel ◽  
Interleukin 8 ◽  
Epithelial Morphogenesis ◽  
Pcr Analysis ◽  
Protein Levels ◽  
Complex Interactions

Epithelial morphogenesis is dependent upon a variety of factors, many of which involve complex interactions between cells and their surrounding environments. We analyzed the patterns of differential gene expression associated with Madin-Darby canine kidney (MDCK) renal epithelial cells grown within a collagen gel in three-dimensional (3D) culture compared with those grown atop a collagen gel in two-dimensional (2D) culture. Under these conditions, MDCK cells spontaneously formed either hollow spherical cysts or flat monolayer sheets, respectively. Microarray analysis of gene expression revealed a twofold or greater expression difference in 732 gene sets from MDCK cysts compared with monolayers (false discovery rate or FDR-adjusted P values <0.05). Interleukin-8 (IL-8) was reproducibly found to be among the genes whose expression was most dramatically upregulated, and this behavior was verified through real-time PCR analysis. The level of IL-8 protein expression was significantly increased in 3D MDCK cultures compared with that detected in cells in 2D culture. Hepatocyte growth factor (HGF) induces MDCK cells in 3D culture to form linear tubule-like structures. We found that HGF stimulation caused MDCK cells in 3D culture to decrease the expression of IL-8 at both the mRNA and protein levels. Furthermore, the addition of recombinant IL-8 to HGF-stimulated 3D MDCK cultures was sufficient to partially reverse the tubulogenic effects of HGF, resulting in the formation of cystic structures. These data suggest that IL-8 participates in the formation of cystic structures by MDCK cells in 3D culture and that HGF may stimulate tubulogenesis through the suppression of IL-8.

scholarly journals HGF-induced serine 897 phosphorylation of EphA2 regulates epithelial morphogenesis of MDCK cells in 3D culture

Development ◽  
2015 ◽  
Vol 142 (11) ◽  
pp. e1105-e1105
Author(s):  
K. Harada ◽  
M. Negishi ◽  
H. Katoh
Keyword(s):  
Mdck Cells ◽  
3D Culture ◽  
Epithelial Morphogenesis

scholarly journals The Transforming Rho Family GTPase Wrch-1 Disrupts Epithelial Cell Tight Junctions and Epithelial Morphogenesis

2008 ◽  
Vol 29 (4) ◽  
pp. 1035-1049 ◽  
Author(s):  
Donita C. Brady ◽  
Jamie K. Alan ◽  
James P. Madigan ◽  
Alan S. Fanning ◽  
Adrienne D. Cox
Keyword(s):  
Epithelial Cell ◽  
Cell Polarity ◽  
Rho Gtpase ◽  
Mdck Cells ◽  
3D Culture ◽  
Cell Polarization ◽  
Epithelial Morphogenesis ◽  
Detectable Effect ◽  
Dependent Manner ◽  
Actin Organization

ABSTRACT Wrch-1, an atypical and transforming Rho GTPase, regulates cellular activities including proliferation and actin organization, but its functions and effectors remain poorly characterized. We show here that Wrch-1 distributes along the apical and basolateral membranes in MDCK cells and binds the cell polarity protein Par6 in a GTP-dependent manner. Activated Wrch-1 negatively regulates the kinetics of tight junction (TJ) assembly during epithelial cell polarization but has no detectable effect on overall cell polarity in confluent monolayers. It also causes a dramatic cytoskeletal reorganization and multilayering in cells grown in two-dimensional culture and disrupts cystogenesis of cells grown in three-dimensional (3D) culture. Similarly, short hairpin RNA-mediated knockdown of Wrch-1 perturbs cystogenesis in 3D culture, suggesting that tight regulation of Wrch-1 activity is necessary for normal epithelial morphogenesis. A weakly transforming effector domain mutant of activated Wrch-1 that inhibits Par6 binding abrogates the ability of Wrch-1 to disrupt TJ formation, actin organization, and epithelial morphogenesis. We hypothesize that Wrch-1-induced morphological and growth transformation may occur in part through Par6-mediated disruption of TJs and actin organization.

scholarly journals Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition

2017 ◽  
Vol 28 (8) ◽  
pp. 1088-1100 ◽  
Author(s):  
Lynne A. Lapierre ◽  
Elizabeth H. Manning ◽  
Kenya M. Mitchell ◽  
Cathy M. Caldwell ◽  
James R. Goldenring
Keyword(s):  
Mdck Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Cyst Formation ◽  
Epithelial Polarity ◽  
Lateral Membrane ◽  
Low Calcium ◽  
Single Lumen ◽  
Apical Junctions ◽  
Temporal Localization

MARK2 regulates the establishment of polarity in Madin–Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral membrane before pS227-FIP2 arrival. Later in recovery, Eps15 and pS227-FIP2 colocalized at the lateral membrane. In MDCK cells expressing the pseudophosphorylated FIP2 mutant FIP2(S227E), during recovery from low calcium, Eps15 was trapped and never localized to the lateral membrane. Mutation of any of the three NPF domains within GFP-FIP2(S227E) rescued Eps15 localization at the lateral membrane and reestablished single-lumen cyst formation in GFP-FIP2(S227E)–expressing cells in three-dimensional (3D) culture. Whereas expression of GFP-FIP2(S227E) induced the loss of E-cadherin and occludin, mutation of any of the NPF domains of GFP-FIP2(S227E) reestablished both proteins at the apical junctions. Knockdown of Eps15 altered the spatial and temporal localization of pS227-FIP2 and also elicited formation of multiple lumens in MDCK 3D cysts. Thus an interaction of Eps15 and pS227-FIP2 at the appropriate time and location in polarizing cells is necessary for proper establishment of epithelial polarity.

Mutant cadherin affects epithelial morphogenesis and invasion, but not transformation

2001 ◽  
Vol 114 (6) ◽  
pp. 1237-1246 ◽  
Author(s):  
M.L. Troxell ◽  
D.J. Loftus ◽  
W.J. Nelson ◽  
J.A. Marrs
Keyword(s):  
Tumor Progression ◽  
Mdck Cells ◽  
Collagen Gel ◽  
Collagen Matrix ◽  
Soft Agar ◽  
Epithelial Morphogenesis ◽  
Adhesion System ◽  
Severe Impairment ◽  
Hepatocyte Growth ◽  
E Cadherin

MDCK cells were engineered to reversibly express mutant E-cadherin protein with a large extracellular deletion. Mutant cadherin overexpression reduced the expression of endogenous E- and K-cadherins in MDCK cells to negligible levels, resulting in decreased cell adhesion. Despite severe impairment of the cadherin adhesion system, cells overexpressing mutant E-cadherin formed fluid-filled cysts in collagen gel cultures and responded to hepatocyte growth factor/scatter factor (HGF/SF) that induced cellular extension formation with a frequency similar to that of control cysts. However, cells were shed from cyst walls into the lumen and into the collagen matrix prior to and during HGF/SF induced tubule extension. Despite the propensity for cell dissociation, MDCK cells lacking cadherin adhesion molecules were not capable of anchorage-independent growth in soft agar and cell proliferation rate was not affected. Thus, cadherin loss does not induce transformation, despite inducing an invasive phenotype, a later stage of tumor progression. These experiments are especially relevant to tumor progression in cells with altered E-cadherin expression, particularly tumor samples with identified E-cadherin extracellular domain genomic mutations.

scholarly journals LGN regulates mitotic spindle orientation during epithelial morphogenesis

2010 ◽  
Vol 189 (2) ◽  
pp. 275-288 ◽  
Author(s):  
Zhen Zheng ◽  
Huabin Zhu ◽  
Qingwen Wan ◽  
Jing Liu ◽  
Zhuoni Xiao ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Mdck Cells ◽  
Apical Cell ◽  
Cell Polarization ◽  
Epithelial Morphogenesis ◽  
Spindle Orientation ◽  
Cell Cortex ◽  
Dividing Cells ◽  
Cortical Localization ◽  
Lateral Cell

Coordinated cell polarization and mitotic spindle orientation are thought to be important for epithelial morphogenesis. Whether spindle orientation is indeed linked to epithelial morphogenesis and how it is controlled at the molecular level is still unknown. Here, we show that the NuMA- and Gα-binding protein LGN is required for directing spindle orientation during cystogenesis of MDCK cells. LGN localizes to the lateral cell cortex, and is excluded from the apical cell cortex of dividing cells. Depleting LGN, preventing its cortical localization, or disrupting its interaction with endogenous NuMA or Gα proteins all lead to spindle misorientation and abnormal cystogenesis. Moreover, artificial mistargeting of endogenous LGN to the apical membrane results in a near 90° rotation of the spindle axis and profound cystogenesis defects that are dependent on cell division. The normal apical exclusion of LGN during mitosis appears to be mediated by atypical PKC. Thus, cell polarization–mediated spatial restriction of spindle orientation determinants is critical for epithelial morphogenesis.

scholarly journals Fibrocystin in MDCK cells: impact on cell adhesion and epithelial morphogenesis

2015 ◽  
Vol 2 (Suppl 1) ◽  
pp. A26 ◽  
Author(s):  
Birga Sötje ◽  
Dieter Haffner ◽  
Wolfgang H Ziegler
Keyword(s):  
Cell Adhesion ◽  
Mdck Cells ◽  
Epithelial Morphogenesis

The role of collagen reorganization on mammary epithelial morphogenesis in a 3D culture model

Biomaterials ◽  
2010 ◽  
Vol 31 (13) ◽  
pp. 3622-3630 ◽  
Author(s):  
Eugen Dhimolea ◽  
Maricel V. Maffini ◽  
Ana M. Soto ◽  
Carlos Sonnenschein
Keyword(s):  
3D Culture ◽  
Mammary Epithelial ◽  
Epithelial Morphogenesis ◽  
Culture Model ◽  
3D Culture Model

scholarly journals Different responses in transformation of MDCK cells in 2D and 3D culture by v-Src as revealed by microarray techniques, RT-PCR and functional assays

2010 ◽  
Vol 90 (6) ◽  
pp. 915-928 ◽  
Author(s):  
Mira Töyli ◽  
Linda Rosberg-Kulha ◽  
Janne Capra ◽  
Jussi Vuoristo ◽  
Sinikka Eskelinen
Keyword(s):  
Mdck Cells ◽  
3D Culture ◽  
Rt Pcr ◽  
Functional Assays ◽  
2D And 3D
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