Constitutively active mutant GS alpha (G225T) and null-mutant G alpha i-2 (G203T) induce primitive endoderm from stem cells

1995 ◽  
Vol 268 (6) ◽  
pp. C1460-C1466 ◽  
Author(s):  
P. Gao ◽  
D. C. Watkins ◽  
C. C. Malbon
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Control Point ◽  
Null Mutant ◽  
Primitive Endoderm ◽  
Mouse Development ◽  
Differentiation Markers ◽  
Early Mouse ◽  
Gs Alpha ◽  
Constitutively Active

In F9 teratocarcinoma stem cells, retinoic acid induces a primitive endoderm-like phenotype and a sharp decline in G alpha i-2, a response mimicked by expression of RNA antisense to G alpha i-2 in the absence of this morphogen (D. C. Watkins, G. L. Johnson, and C. C. Malbon. Science Wash. DC 258: 1373-1375, 1992). The role of the GS alpha/G alpha i-2 axis in cellular differentiation was explored. In the absence of retinoic acid, F9 stem cells stably expressing a constitutively active mutant of GS alpha (G225T) progressed to the primitive endoderm phenotype, as judged by morphological and differentiation markers, such as tissue plasminogen activator. Although elevated in cells expressing G225T GS alpha, adenosine 3',5'-cyclic monophosphate does not mimic retinoic acid action and alone fails to induce stem cells to primitive endoderm. In the absence of retinoic acid, expression of a null mutant of G alpha i-2 (G203T) also induced stem cells to primitive endoderm. These observations establish G proteins in the GS alpha/G alpha i-2 axis as a control point for regulating progression to primitive endoderm independent of adenylate cyclase, in the present study's model of early mouse development.

scholarly journals Live imaging of X chromosome reactivation dynamics in early mouse development can discriminate naïve from primed pluripotent stem cells

Development ◽  
2016 ◽  
Vol 143 (16) ◽  
pp. 2958-2964 ◽  
Author(s):  
Shin Kobayashi ◽  
Yusuke Hosoi ◽  
Hirosuke Shiura ◽  
Kazuo Yamagata ◽  
Saori Takahashi ◽  
...  
Keyword(s):  
Stem Cells ◽  
X Chromosome ◽  
Pluripotent Stem Cells ◽  
Live Imaging ◽  
Mouse Development ◽  
Early Mouse

scholarly journals Bmi1 facilitates primitive endoderm formation by stabilizing Gata6 during early mouse development

2012 ◽  
Vol 26 (13) ◽  
pp. 1445-1458 ◽  
Author(s):  
F. Lavial ◽  
S. Bessonnard ◽  
Y. Ohnishi ◽  
A. Tsumura ◽  
A. Chandrashekran ◽  
...  
Keyword(s):  
Primitive Endoderm ◽  
Mouse Development ◽  
Early Mouse

scholarly journals A novel role for Gab2 in bFGF-mediated cell survival during retinoic acid–induced neuronal differentiation

2005 ◽  
Vol 170 (2) ◽  
pp. 305-316 ◽  
Author(s):  
Yingwei Mao ◽  
Angel W.-M. Lee
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Neuronal Differentiation ◽  
Embryonic Stem ◽  
Akt Activation ◽  
Important Player ◽  
Ec Cells ◽  
Phosphoinositide 3 Kinase ◽  
Induced Apoptosis ◽  
Constitutively Active

Gab proteins amplify and integrate signals stimulated by many growth factors. In culture and animals, retinoic acid (RA) induces neuronal differentiation. We show that Gab2 expression is detected in neurons in three models of neuronal differentiation: embryonic carcinoma (EC) stem cells, embryonic stem cells, and primary neural stem cells (NSCs). RA treatment induces apoptosis, countered by basic FGF (bFGF). In EC cells, Gab2 silencing results in hypersensitivity to RA-induced apoptosis and abrogates the protection by bFGF. Gab2 suppression reduces bFGF-dependent activation of AKT but not ERK, and constitutively active AKT, but not constitutively active MEK1, reverses the hypersensitization. Thus, Gab2-mediated AKT activation is required for bFGF's protection. Moreover, Gab2 silencing impairs the differentiation of EC cells to neurons. Similarly, in NSCs, Gab2 suppression reduces bFGF-dependent proliferation as well as neuronal survival and production upon differentiation. Our findings provide the first evidence that Gab2 is an important player in neural differentiation, partly by acting downstream of bFGF to mediate survival through phosphoinositide 3 kinase–AKT.

scholarly journals Functional evaluation of transposable elements as enhancers in mouse embryonic and trophoblast stem cells

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Christopher D Todd ◽  
Özgen Deniz ◽  
Darren Taylor ◽  
Miguel R Branco
Keyword(s):  
Stem Cells ◽  
Gene Regulation ◽  
Transposable Elements ◽  
Specific Gene ◽  
Functional Evaluation ◽  
Mouse Development ◽  
Trophoblast Stem Cells ◽  
Trophoblast Stem ◽  
Gene Regulatory ◽  
Early Mouse

Transposable elements (TEs) are thought to have helped establish gene regulatory networks. Both the embryonic and extraembryonic lineages of the early mouse embryo have seemingly co-opted TEs as enhancers, but there is little evidence that they play significant roles in gene regulation. Here we tested a set of long terminal repeat TE families for roles as enhancers in mouse embryonic and trophoblast stem cells. Epigenomic and transcriptomic data suggested that a large number of TEs helped to establish tissue-specific gene expression programmes. Genetic editing of individual TEs confirmed a subset of these regulatory relationships. However, a wider survey via CRISPR interference of RLTR13D6 elements in embryonic stem cells revealed that only a minority play significant roles in gene regulation. Our results suggest that a subset of TEs are important for gene regulation in early mouse development, and highlight the importance of functional experiments when evaluating gene regulatory roles of TEs.

scholarly journals A genetic dissection of the retinoid signalling pathway in the mouse

1999 ◽  
Vol 58 (3) ◽  
pp. 609-613 ◽  
Author(s):  
Manuel Mark ◽  
Norbert B. Ghyselinck ◽  
Olivia Wendling ◽  
Valérie Dupé ◽  
Bénédicte Mascrez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Homologous Recombination ◽  
Embryonic Stem ◽  
Signalling Pathway ◽  
Retinoic Acid Receptors ◽  
Genetic Dissection ◽  
Null Mutant ◽  
Retinoic Acid Signalling ◽  
Null Mutant Mice

To determine the functions of retinoic acid receptors RAR and RXR, we have systematically knocked-out their genes by homologous recombination in the embryonic stem cells and generated null-mutant mice. This approach has allowed us to perform a genetic dissection of the retinoic acid signalling pathway.

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.

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