RhoA/ROCK and Cdc42 regulate cell‐cell contact and N‐cadherin protein level during neurodetermination of P19 embryonal stem cells

Laplante, Isabel; Béliveau, Richard; Paquin, Joanne

doi:10.1002/neu.20036

Cited by 34 publications

(25 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…al. demonstrated that RhoA is implicated during neurodetermination, where it influences cell-cell contact and cadherin levels [56]. They used the P19 cell model of neuronal differentiation to show that RhoA affects cadherin protein level and cell-cell contacts during neuroinduction.…”

Section: Discussionmentioning

confidence: 99%

Role of the microtubule-targeting drug vinflunine on cell-cell adhesions in bladder epithelial tumour cells

et al. 2014

View full text Add to dashboard Cite

BackgroundVinflunine (VFL) is a microtubule-targeting drug that suppresses microtubule dynamics, showing anti-metastatic properties both in vitro and in living cancer cells. An increasing body of evidence underlines the influence of the microtubules dynamics on the cadherin-dependent cell-cell adhesions. E-cadherin is a marker of epithelial-to-mesenchymal transition (EMT) and a tumour suppressor; its reduced levels in carcinoma are associated with poor prognosis. In this report, we investigate the role of VFL on cell-cell adhesions in bladder epithelial tumour cells.MethodsHuman bladder epithelial tumour cell lines HT1376, 5637, SW780, T24 and UMUC3 were used to analyse cadherin-dependent cell-cell adhesions under VFL treatment. VFL effect on growth inhibition was measured by using a MTT colorimetric cell viability assay. Western blot, immunofluorescence and transmission electron microscopy analyses were performed to assess the roles of VFL effect on cell-cell adhesions, epithelial-to-mesenchymal markers and apoptosis. The role of the proteasome in controlling cell-cell adhesion was studied using the proteasome inhibitor MG132.ResultsWe show that VFL induces cell death in bladder cancer cells and activates epithelial differentiation of the remaining living cells, leading to an increase of E-cadherin-dependent cell-cell adhesion and a reduction of mesenchymal markers, such as N-cadherin or vimentin. Moreover, while E-cadherin is increased, the levels of Hakai, an E3 ubiquitin-ligase for E-cadherin, were significantly reduced in presence of VFL. In 5637, this reduction on Hakai expression was blocked by MG132 proteasome inhibitor, indicating that the proteasome pathway could be one of the molecular mechanisms involved in its degradation.ConclusionsOur findings underscore a critical function for VFL in cell-cell adhesions of epithelial bladder tumour cells, suggesting a novel molecular mechanism by which VFL may impact upon EMT and metastasis.

show abstract

Section: Discussionmentioning

confidence: 99%

Role of the microtubule-targeting drug vinflunine on cell-cell adhesions in bladder epithelial tumour cells

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Skeletal muscle cells are also generated with cardiomyocytes in these cultures [1,10]. It is worthy of note that exposure of P19 cells to higher RA concentration (10 −6 M) over the aggregation period generates neurons but not muscle cells [1,[15][16][17]. Interestingly application of such high RA concentration at midterm instead of beginning of the cell aggregation period followed by exposure to T3 and insulin was shown to induce differentiation of ES cells into adipocytes and skeletomyocytes, and perhaps into cardiomyocytes also although generation of this last cell type was not demonstrated [18].…”

Section: Immunocytochemistrymentioning

confidence: 99%

“…Dye retained by cells was extracted with isopropanol and measured by absorbance at 510 nm. Cultures were then counterstained with Crystal violet (absorbance at 595 nm [16]) to correct Oil-Red-O values for variations in cell culture densities.…”

Section: Oil-red-o Stainingmentioning

confidence: 99%

Skeletal and Cardiac Myogenesis Accompany Adipogenesis in P19 Embryonal Stem Cells

Bouchard

Paquin

2009

Stem Cells and Development

Self Cite

View full text Add to dashboard Cite

P19 embryonic carcinoma cells resemble normal embryonic stem (ES) cells. They generate cardiac and skeletal myocytes in response to retinoic acid (RA) or oxytocin (OT). RA treatment followed by exposure to triiodothyronine (T3) and insulin induces ES cells differentiation into adipocytes and skeletomyocytes. On the other hand, OT (10(-7) M) was reported to inhibit 3T3 preadipocyte maturation. The present work was undertaken to determine whether P19 cells have an adipogenic potential that could be affected by OT. Cells were treated with RA (10(-6) M)/T3+insulin (adipogenic protocol) or 10(-7) M OT (cardiomyogenic protocol), and analyzed by polymerase chain reaction, immunotechniques, and cytochemistry. Oil-Red-O staining and expression of peroxisome proliferator-activated receptor-gamma (PPARgamma) and aP2 indicated the generation of adipocytes in cultures submitted to the adipogenic protocol. Contracting cells were also generated. Cells positive for sarcomeric actinin and negative for cardiac troponin inhibitor (cTpnI) indicated generation of skeletomyocytes, and cTpnI positive cells revealed generation of cardiomyocytes. Levels of cTpnI and of the skeletal marker MyoD were almost similar in both protocols, whereas no Oil-Red-O staining was associated with the cardiomyogenic protocol. Addition of 10(-7) M OT to the adipogenic protocol did not affect Oil-Red-O staining and PPARgamma expression. Interestingly, Oct3/4 pluripotency marker disappeared in the adipogenic protocol but remained expressed in the cardiomyogenic one. P19 cells thus have an adipogenic potential non affected by 10(-7) M OT. RA/T3+insulin combination generates a larger spectrum of mesodermal cell derivatives and is a more potent morphogenic treatment than OT. P19 cells could help investigating mechanisms of cell fate decision during development.

show abstract

“…Because RhoA signaling is necessary for cadherindependent cell-cell contact formation and maintenance in flies and in different mammalian cell types (Sahai and Marshall, 2002;Laplante et al, 2004;Homem and Peifer, 2008) we investigated whether the loss of RhoA affected cell-cell adhesion in the spinal cord neuroepithelium of mutants. We analyzed the distribution of the AJ components N-cadherin, ␤-catenin, and actin at the apical side of NEPs lining the lumen of the neural tube by immunofluorescence (Fig.…”

Section: Rhoa Is Required For the Maintenance Of N-cadherin-based Celmentioning

confidence: 99%

The Small GTPase RhoA Is Required to Maintain Spinal Cord Neuroepithelium Organization and the Neural Stem Cell Pool

Herzog¹,

Loetscher²,

Hengel³

et al. 2011

J. Neurosci.

View full text Add to dashboard Cite

The regulation of adherens junctions (AJs) is critical for multiple events during CNS development, including the formation and maintenance of the neuroepithelium. We have addressed the role of the small GTPase RhoA in the developing mouse nervous system using tissue-specific conditional gene ablation. We show that, in the spinal cord neuroepithelium, RhoA is essential to localize N-cadherin and ␤-catenin to AJs and maintain apical-basal polarity of neural progenitor cells. Ablation of RhoA caused the loss of AJs and severe abnormalities in the organization of cells within the neuroepithelium, including decreased neuroepithelial cell proliferation and premature cell-cycle exit, reduction of the neural stem cell pool size, and the infiltration of neuroepithelial cells into the lumen of the ventricle. We also show that, in the absence of RhoA, its effector, mammalian diaphanous-related formin1 (mDia1), does not localize to apical AJs in which it likely stabilizes intracellular adhesion by promoting local actin polymerization and microtubule organization. Furthermore, expressing a dominant-negative form of mDia1 in neural stem/progenitor cells results in a similar phenotype compared with that of the RhoA conditional knock-out, namely the loss of AJs and apical polarity. Together, our data show that RhoA signaling is necessary for AJ regulation and for the maintenance of mammalian neuroepithelium organization preventing precocious cell-cycle exit and differentiation.

show abstract

RhoA/ROCK and Cdc42 regulate cell‐cell contact and N‐cadherin protein level during neurodetermination of P19 embryonal stem cells

Cited by 34 publications

References 65 publications

Role of the microtubule-targeting drug vinflunine on cell-cell adhesions in bladder epithelial tumour cells

Role of the microtubule-targeting drug vinflunine on cell-cell adhesions in bladder epithelial tumour cells

Skeletal and Cardiac Myogenesis Accompany Adipogenesis in P19 Embryonal Stem Cells

The Small GTPase RhoA Is Required to Maintain Spinal Cord Neuroepithelium Organization and the Neural Stem Cell Pool

Contact Info

Product

Resources

About