scholarly journals Frizzled gene expression and negative regulation of canonical WNT–β-catenin signaling in mouse F9 teratocarcinoma cells

2017 ◽  
Vol 95 (2) ◽  
pp. 251-262 ◽  
Author(s):  
Gregory Golenia ◽  
Mohamed I. Gatie ◽  
Gregory M. Kelly
Keyword(s):  
Feedback Loop ◽  
Negative Regulation ◽  
Negative Feedback Loop ◽  
Primitive Endoderm ◽  
F9 Cells ◽  
Knock Down ◽  
Teratocarcinoma Cells ◽  
Parietal Endoderm ◽  
F9 Teratocarcinoma ◽  
Canonical Wnt

Mouse F9 cells differentiate into primitive endoderm (PrE) following the activation of the canonical WNT–β-catenin pathway. The upregulation of Wnt6 and activation of β-catenin–TCF–LEF-dependent transcription is known to accompany differentiation, but the Frizzled (FZD) receptor responsible for transducing the WNT6 signal is not known. Eight of the 10 Fzd genes were found to be expressed in F9 cells, with Fzd7 being the most highly expressed, and chosen for further analysis. To alter steady-state Fzd7 levels and test the effect this has on differentiation, siRNA and overexpression approaches were used to knock-down and ectopically express the Fzd7 message, respectively. siRNA knock-down of Fzd7 resulted in reduced DAB2 levels, and the overexpression activated a TCF–LEF reporter, but neither approach affected differentiation. Our focus turned to how canonical WNT6 signaling was attenuated to allow PrE cells to form parietal endoderm (PE). Dkk1, encoding a WNT antagonist, was examined and results showed that its expression increased in F9 cells treated with retinoic acid (RA) or overexpressing Wnt6. F9 cells overexpressing human DKK1 or treated with DKK1-conditioned medium and then treated with RA failed to differentiate, indicating that a negative feedback loop involving WNT6 and DKK1 attenuates canonical WNT–β-catenin signaling, thereby allowing PE cells to differentiate.

scholarly journals Expression of fibroblast growth factor by F9 teratocarcinoma cells as a function of differentiation.

1989 ◽  
Vol 108 (6) ◽  
pp. 2467-2476 ◽  
Author(s):  
S J Braunhut ◽  
L J Gudas ◽  
T Kurokawa ◽  
J Sasse ◽  
P A D'Amore
Keyword(s):  
Stem Cells ◽  
Cdna Probe ◽  
Biologically Active ◽  
Endothelial Cell Proliferation ◽  
Fibroblast Growth ◽  
Basic Fgf ◽  
F9 Cells ◽  
Parietal Endoderm ◽  
F9 Teratocarcinoma ◽  
Proliferation Assays

F9 teratocarcinoma stem cells treated with retinoic acid (RA) and dibutyryl cAMP (but2 cAMP) differentiate into embryonic parietal endoderm. Using heparin-affinity chromatography, endothelial cell proliferation assays, immunoprecipitation, and Western analysis with antibodies specific for acidic and basic fibroblast growth factors (FGFs), we detected biologically active FGF in F9 cells only after differentiation. A bovine basic FGF cDNA probe hybridized to 2.2-kb mRNAs in both F9 stem and parietal endoderm cells and to a 3.8-kb mRNA in F9 stem cells. A genomic DNA probe for acidic FGF hybridized to a 5.8-6.0-kb mRNA in both F9 stem and parietal endoderm cells, and to a 6.0-6.3-kb mRNA only in parietal endoderm cells. Although these FGF mRNAs were present in the stem cells, we could find no evidence that F9 stem cells synthesized FGFs, whereas differentiated F9 cells synthesized both acidic and basic FGF-like proteins. We conclude that biologically active factors with properties characteristic of acidic and basic FGF are expressed by F9 parietal endoderm cells after differentiation. Differentiating embryonic parietal endoderm thus may serve as a source of FGF molecules in the developing blastocyst, where these factors appear to play a central role in subsequent embryogenesis.

scholarly journals Retinoic acid induces liver/bone/kidney-type alkaline phosphatase gene expression in F9 teratocarcinoma cells

1991 ◽  
Vol 274 (3) ◽  
pp. 673-678 ◽  
Author(s):  
M Gianni ◽  
M Studer ◽  
G Carpani ◽  
M Terao ◽  
E Garattini
Keyword(s):  
Alkaline Phosphatase ◽  
Retinoic Acid ◽  
De Novo ◽  
F9 Cells ◽  
Alp Activity ◽  
Mrna Induction ◽  
All Trans Retinoic Acid ◽  
Teratocarcinoma Cells ◽  
F9 Teratocarcinoma ◽  
De Novo Protein Synthesis

All-trans retinoic acid (RA) induces alkaline phosphatase (ALP) activity by 3-8-fold in murine F9 teratocarcinoma cells, in parallel with their differentiation towards primitive endoderm. The elevation of ALP activity is associated with increases in the amounts of liver/bone/kidney-type ALP protein and the respective transcript. These effects of RA are due to activation of ALP gene transcription rather than to an increase in the half-life of the mRNA. Induction of ALP mRNA does not require de novo protein synthesis, since it is not blocked by treatment with cycloheximide. Dibutyryl cyclic AMP, which is known to induce further differentiation of F9 cells from the primitive to the parietal endoderm, blocks the induction of ALP mRNA by RA.

Reversible Interconversion between Primitive Endoderm- and Parietal Endoderm-Like F9 Cells Demonstrated by mRNAs Expression1

1987 ◽  
Vol 102 (2) ◽  
pp. 385-392 ◽  
Author(s):  
Kiyoshi MIKI ◽  
Etsuro SUGIMOTO ◽  
Yasuo KITAGAWA
Keyword(s):  
Primitive Endoderm ◽  
F9 Cells ◽  
Parietal Endoderm

Induction of alpha-fetoprotein synthesis in differentiating F9 teratocarcinoma cells is accompanied by a genome-wide loss of DNA methylation

1984 ◽  
Vol 4 (5) ◽  
pp. 898-907
Author(s):  
P R Young ◽  
S M Tilghman
Keyword(s):  
Dna Methylation ◽  
Dna Methyltransferase ◽  
Alpha Fetoprotein ◽  
Restriction Enzyme Digestion ◽  
Visceral Endoderm ◽  
F9 Cells ◽  
Genome Wide ◽  
Teratocarcinoma Cells ◽  
A Genome ◽  
F9 Teratocarcinoma

F9 teratocarcinoma cells can be grown as monolayers or aggregates, and upon treatment with retinoic acid they will differentiate into parietal or visceral endoderm, respectively. Visceral endoderm specifically synthesizes alpha-fetoprotein and albumin mRNAs, which are not found in parietal endoderm. In contrast, both endoderms produce enhanced levels of the major histocompatibility antigen (H2) mRNA compared with F9 cells. F9 cells contain highly methylated DNA as judged by restriction enzyme digestion. However, upon differentiation into visceral endoderm, there is a genome-wide loss of methylation in induced, silent, and constitutively expressed genes. Experiments in which methylation loss is induced via the methyltransferase inhibitor 5-azacytidine result in no induction of alpha-fetoprotein mRNA and no morphological differentiation, suggesting that methylation loss alone is not sufficient to induce the visceral endoderm phenotype. Likewise, 5-azacytidine treatment of differentiated cells does not result in enhanced expression of alpha-fetoprotein mRNA. However, the patterns of loss of DNA methylation at all sites examined after differentiation or 5-azacytidine treatment were remarkably similar, suggesting that the two occur by a similar mechanism, the inhibition of DNA methyltransferase activity. These results argue that the specificity for methylation loss at a given site is an inherent property of aggregated F9 cell chromatin. This system provides a model for studying a tissue-specific change in DNA methylation upon differentiation.

scholarly journals Induction of alpha-fetoprotein synthesis in differentiating F9 teratocarcinoma cells is accompanied by a genome-wide loss of DNA methylation.

1984 ◽  
Vol 4 (5) ◽  
pp. 898-907 ◽  
Author(s):  
P R Young ◽  
S M Tilghman
Keyword(s):  
Dna Methylation ◽  
Dna Methyltransferase ◽  
Alpha Fetoprotein ◽  
Restriction Enzyme Digestion ◽  
Visceral Endoderm ◽  
F9 Cells ◽  
Genome Wide ◽  
Teratocarcinoma Cells ◽  
A Genome ◽  
F9 Teratocarcinoma

F9 teratocarcinoma cells can be grown as monolayers or aggregates, and upon treatment with retinoic acid they will differentiate into parietal or visceral endoderm, respectively. Visceral endoderm specifically synthesizes alpha-fetoprotein and albumin mRNAs, which are not found in parietal endoderm. In contrast, both endoderms produce enhanced levels of the major histocompatibility antigen (H2) mRNA compared with F9 cells. F9 cells contain highly methylated DNA as judged by restriction enzyme digestion. However, upon differentiation into visceral endoderm, there is a genome-wide loss of methylation in induced, silent, and constitutively expressed genes. Experiments in which methylation loss is induced via the methyltransferase inhibitor 5-azacytidine result in no induction of alpha-fetoprotein mRNA and no morphological differentiation, suggesting that methylation loss alone is not sufficient to induce the visceral endoderm phenotype. Likewise, 5-azacytidine treatment of differentiated cells does not result in enhanced expression of alpha-fetoprotein mRNA. However, the patterns of loss of DNA methylation at all sites examined after differentiation or 5-azacytidine treatment were remarkably similar, suggesting that the two occur by a similar mechanism, the inhibition of DNA methyltransferase activity. These results argue that the specificity for methylation loss at a given site is an inherent property of aggregated F9 cell chromatin. This system provides a model for studying a tissue-specific change in DNA methylation upon differentiation.

c-myc regulation during retinoic acid-induced differentiation of F9 cells is posttranscriptional and associated with growth arrest

1986 ◽  
Vol 6 (2) ◽  
pp. 518-524
Author(s):  
M Dean ◽  
R A Levine ◽  
J Campisi
Keyword(s):  
Retinoic Acid ◽  
Cyclic Amp ◽  
Posttranscriptional Regulation ◽  
Growth Arrest ◽  
Mrna Levels ◽  
F9 Cells ◽  
Induction Of Differentiation ◽  
Parietal Endoderm ◽  
Arrest Growth ◽  
F9 Teratocarcinoma

We have shown that c-myc mRNA levels decrease more than 20-fold when F9 teratocarcinoma stem cells are induced to arrest growth and terminally differentiate to parietal endoderm after exposure to retinoic acid and cyclic AMP (Campisi et al., Cell 36:241-247, 1984). Here, we demonstrate that although growth arrest and full expression of the differentiated phenotype required about 3 days, c-myc mRNA declined abruptly between 8 and 16 h after the addition of retinoic acid and cyclic AMP. The decline was independent of cyclic AMP. We found little or no change in the level of c-myc transcription during differentiation, although two other genes showed marked transcriptional regulation. Thus, decreased c-myc mRNA is a consequence of very early posttranscriptional regulation directed by retinoic acid. Differentiation was not fundamental to this regulation. We have shown that sodium butyrate blocks expression of the differentiated phenotype if added within 8 h of retinoic acid and cyclic AMP (Levine et al., Dev. Biol. 105:443-450, 1984). However, butyrate did not inhibit the decrease in c-myc mRNA. Furthermore, F9 cells partially arrested growth without differentiating when grown in isoleucine-deficient medium. Under these conditions, c-myc mRNA levels also declined. Our results suggest that induction of differentiation-specific genes may be under retinoic acid-mediated control dissimilar from that responsible for the decay of c-myc mRNA. In addition, they raise the possibility that growth arrest may be initiated by reduced c-myc expression.

A role for p21 (WAF1) in the cAMP-dependent differentiation of F9 teratocarcinoma cells into parietal endoderm

2005 ◽  
Vol 304 (1) ◽  
pp. 293-304 ◽  
Author(s):  
B DRDOVA ◽  
J VACHTENHEIM
Keyword(s):  
Teratocarcinoma Cells ◽  
Parietal Endoderm ◽  
P21 Waf1 ◽  
F9 Teratocarcinoma

Coordinate Gα13 and Wnt6-β-catenin signaling in F9 embryonal carcinoma cells is required for primitive endoderm differentiation

2009 ◽  
Vol 87 (4) ◽  
pp. 567-580 ◽  
Author(s):  
Roman Krawetz ◽  
Gregory M. Kelly
Keyword(s):  
Retinoic Acid ◽  
Epithelial To Mesenchymal Transition ◽  
Embryonal Carcinoma ◽  
Embryonal Carcinoma Cell ◽  
Primitive Endoderm ◽  
F9 Cells ◽  
Mesenchymal Transition ◽  
Signaling Axis ◽  
Parietal Endoderm ◽  
Constitutively Active

The mouse F9 embryonal carcinoma cell line is ideally suited to study the epithelial-to-mesenchymal transition accompanying the differentiation of primitive to parietal extraembryonic endoderm. In F9 cells, the application of exogenous agents including retinoic acid or activation of signal transduction cascades downstream of G-proteins triggers widespread changes in gene expression and leads to the formation of primitive endoderm. The epithelial-to-mesenchymal transition is completed and parietal endoderm develops as of result of increasing PKA activity in primitive endoderm cells. Expression of a constitutively active form of Gα13(Q226L) is sufficient to induce F9 cells into parietal endoderm and a model is emerging that a signaling axis linking G-protein signaling to RhoA and the ERM protein moesin is required for differentiation. In this study, we found that expression of either p115RhoGEF or a constitutively active, GTPase-deficient form of RhoA(L63) promoted primitive, but not parietal, endoderm formation. The overexpression of Gα13(Q226L) or p115RhoGEF, but not Rho(L63), caused β-catenin to translocate to the nucleus. Surprisingly, the stimulation of the Wnt-β-catenin pathway was accompanied by nuclear β-catenin and primitive endoderm formation, even when a dominant negative was used to block the signaling axis at the level of p115RhoGEF or when ROCK activity was inhibited using the pharmacological agent Y-27632. Together, results indicate that the coordinate signaling by two independent pathways, one involving canonical Wnt-β-catenin activation of target genes and the other with Gα13 signaling to ERM proteins to modulate cytoarchitectural changes, is required during the retinoic acid induced differentiation of F9 cells to primitive endoderm.

Expression of c-fos in parietal endoderm, amnion and differentiating F9 teratocarcinoma cells

1985 ◽  
Vol 30 (1) ◽  
pp. 76-81 ◽  
Author(s):  
Ivor Mason ◽  
David Murphy ◽  
Brigid L.M. Hogan
Keyword(s):  
Teratocarcinoma Cells ◽  
Parietal Endoderm ◽  
F9 Teratocarcinoma
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