scholarly journals LGR4, Not LGR5, Enhances hPSC Hematopoiesis by Facilitating Mesoderm Induction via TGF-Beta Signaling Activation

Cell Reports ◽  
2020 ◽  
Vol 31 (5) ◽  
pp. 107600 ◽  
Author(s):  
Yu Wang ◽  
Hongtao Wang ◽  
Jiaojiao Guo ◽  
Jie Gao ◽  
Mengge Wang ◽  
...  
Keyword(s):  
Mesoderm Induction ◽  
Tgf Beta ◽  
Signaling Activation

scholarly journals Evidence for involvement of activin A and bone morphogenetic protein 4 in mammalian mesoderm and hematopoietic development.

1995 ◽  
Vol 15 (1) ◽  
pp. 141-151 ◽  
Author(s):  
B M Johansson ◽  
M V Wiles
Keyword(s):  
Growth Factor ◽  
Bone Morphogenetic Protein ◽  
Activin A ◽  
Bone Morphogenetic Protein 4 ◽  
Mesoderm Induction ◽  
Dependent Manner ◽  
Tgf Beta ◽  
Mouse Development ◽  
Morphogenetic Protein

Xenopus in vitro studies have implicated both transforming growth factor beta (TGF-beta) and fibroblast growth factor (FGF) families in mesoderm induction. Although members of both families are present during mouse mesoderm formation, there is little evidence for their functional role in mesoderm induction. We show that mouse embryonic stem cells, which resemble primitive ectoderm, can differentiate to mesoderm in vitro in a chemically defined medium (CDM) in the absence of fetal bovine serum. In CDM, this differentiation is responsive to TGF-beta family members in a concentration-dependent manner, with activin A mediating the formation of dorsoanterior-like mesoderm and bone morphogenetic protein 4 mediating the formation of ventral mesoderm, including hematopoietic precursors. These effects are not observed in CDM alone or when TGF-beta 1, -beta 2, or -beta 3, acid FGF, or basic FGF is added individually to CDM. In vivo, at day 6.5 of mouse development, activin beta A RNA is detectable in the decidua and bone morphogenetic protein 4 RNA is detectable in the egg cylinder. Together, our data strongly implicate the TGF-beta family in mammalian mesoderm development and hematopoietic cell formation.

Mesoderm induction in amphibians: the role of TGF-beta 2-like factors

Science ◽  
1988 ◽  
Vol 239 (4841) ◽  
pp. 783-785 ◽  
Author(s):  
F Rosa ◽  
A. Roberts ◽  
D Danielpour ◽  
L. Dart ◽  
M. Sporn ◽  
...  
Keyword(s):  
Mesoderm Induction ◽  
Tgf Beta ◽  
Beta 2

scholarly journals Activin-mediated mesoderm induction requires FGF

Development ◽  
1994 ◽  
Vol 120 (2) ◽  
pp. 453-462 ◽  
Author(s):  
R.A. Cornell ◽  
D. Kimelman
Keyword(s):  
Dominant Negative ◽  
Mesoderm Induction ◽  
Fibroblast Growth ◽  
Fgf Signaling ◽  
Tgf Beta ◽  
Fgf Receptor ◽  
Basic Fgf ◽  
Dominant Negative Form ◽  
Negative Form

The early patterning of mesoderm in the Xenopus embryo requires signals from several intercellular factors, including mesoderm-inducing agents that belong to the fibroblast growth factor (FGF) and TGF-beta families. In animal hemisphere explants (animal caps), basic FGF and the TGF-beta family member activin are capable of converting pre-ectodermal cells to a mesodermal fate, although activin is much more effective at inducing dorsal and anterior mesoderm than is basic FGF. Using a dominant-negative form of the Xenopus type 1 FGF receptor, we show that an FGF signal is required for the full induction of mesoderm by activin. Animal caps isolated from embryos that have been injected with the truncated FGF receptor and cultured with activin do not extend and the induction of some genes, including cardiac actin and Xbra, is greatly diminished, while the induction of other genes, including the head organizer-specific genes gsc and Xlim-1, is less sensitive. These results are consistent with the phenotype of the truncated FGF receptor-injected embryo and imply that the activin induction of mesoderm depends on FGF, with some genes requiring a higher level of FGF signaling than others.

Cellular interpretation of multiple TGF-beta signals: intracellular antagonism between activin/BVg1 and BMP-2/4 signaling mediated by Smads

Development ◽  
1997 ◽  
Vol 124 (22) ◽  
pp. 4467-4480 ◽  
Author(s):  
A.F. Candia ◽  
T. Watabe ◽  
S.H. Hawley ◽  
D. Onichtchouk ◽  
Y. Zhang ◽  
...  
Keyword(s):  
Cell Fate ◽  
Bone Morphogenetic Proteins ◽  
Dominant Negative ◽  
Signaling Cascades ◽  
Mesoderm Induction ◽  
Tgf Beta ◽  
Functional Involvement ◽  
Dominant Negative Variant ◽  
Transcriptional Complex ◽  
Mesoderm Patterning

During early embryogenesis of Xenopus, dorsoventral polarity of the mesoderm is established by dorsalizing and ventralizing agents, which are presumably mediated by the activity of an activin/BVg1-like protein and Bone Morphogenetic Proteins (BMP), respectively. Interestingly, these two TGF-beta subfamilies are found in overlapping regions during mesoderm patterning. This raises the question of how the presumptive mesodermal cells recognize the multiple TGF-beta signals and differentially interpret this information to assign a particular cell fate. In this study, we have exploited the well characterized model of Xenopus mesoderm induction to determine the intracellular interactions between BMP-2/4 and activin/BVg1 signaling cascades. Using a constitutively active BMP-2/4 receptor that transduces BMP-2/4 signals in a ligand-independent fashion, we demonstrate that signals provided by activin/BVg1 and BMP modulate each other's activity and that this crosstalk occurs through intracellular mechanisms. In assays using BMP-2/4 and activin/BVg1-specific reporters, we determined that the specificity of BMP-2/4 and activin/BVg1 signaling is mediated by Smad1 and Smad2, respectively. These Smads should be considered as the mediators of the intracellular antagonism between BMP-2/4 and activin/BVg1 signaling possibly through sequestration of a limited pool of Smad4. Consistent with such a mechanism, Smad4 interacts functionally with both Smad1 and −2 to potentiate their signaling activities, and a dominant negative variant of Smad4 can inhibit both activin/BVg1 and BMP-2/4 mediated signaling Finally, we demonstrate that an activin/BVg1-dependent transcriptional complex contains both Smad2 and Smad4 and thereby provides a physical basis for the functional involvement of both Smads in TGF-beta-dependent transcriptional regulation. Thus, Smad4 plays a central role in synergistically activating activin/BVg1 and BMP-dependent transcription and functions as an intracellular sensor for TGF-beta-related signals.

The biological effects of XTC-MIF: quantitative comparison with Xenopus bFGF

Development ◽  
1990 ◽  
Vol 108 (1) ◽  
pp. 173-183 ◽  
Author(s):  
J.B. Green ◽  
G. Howes ◽  
K. Symes ◽  
J. Cooke ◽  
J.C. Smith
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Biological Effects ◽  
Tissue Type ◽  
Mesoderm Induction ◽  
Tgf Beta ◽  
Signal Molecule ◽  
Animal Pole

Mesoderm in Xenopus and other amphibian embryos is induced by signals from the vegetal hemisphere acting on equatorial or animal hemisphere cells. These signals are diffusible and two classes of candidate signal molecule have been identified: the fibroblast growth factor (FGF) and transforming growth factor beta (TGF-beta) types. In this paper, we compare the effects of cloned Xenopus basic FGF (XbFGF) and electophoretically homogeneous XTC-MIF (a TGF-beta-like factor obtained from a Xenopus cell line) on animal pole explants. We find that they have a similar minimum active concentration (0.1-0.2 ng ml-1) but that, nonetheless, XTC-MIF is at least 40 times more active in inducing muscle. In general, we find that the two factors cause inductions of significantly different characters in terms of tissue type, morphology, gene expression and timing. At low concentrations (0.1-1.0 ng ml-1) both factors induce the differentiation of ‘mesenchyme’ and ‘mesothelium’ as well as blood-like cells. These latter cells do not, however, react with an antibody to Xenopus globin. This raised the possibility that the identification of red blood cells in other studies on mesoderm induction might have been mistaken, but combinations of animal pole regions with ventral vegetal pole regions confirmed that genuine erythrocytes are formed. The identity of the blood-like cells formed in response to the inducing factors remains unknown. At higher concentrations XTC-MIF induces neural tissue, notochord, pronephros and substantial and often segmented muscle. By contrast, XbFGF only induces significant amounts of muscle above 24 ng ml-1 and even then this is much less than that induced by XTC-MIF. For both factors an exposure of less than 30 min is effective. Competence of animal pole cells to respond to XbFGF is completely lost by the beginning of gastrulation (stage 10) while competence to XTC-MIF is detectable until somewhat later (stage 11). Since animal pole tissue is known to be able to respond to the natural inducer at least until stage 10, and perhaps until stage 10.5, this suggests that bFGF cannot be the sole inducer of mesoderm in vivo. Taken together, these results are consistent with XTC-MIF being a dorsoanterior inducer and XbFGF a ventroposterior inducer, suggesting that body pattern is established by the interaction of two types of inducing signal. This model is discussed in view of the qualitative and quantitative differences between the factors.

The Balance of Positive and Negative Effects of TGF-Beta Signaling Regulates the Hemangioblast Development in Human Pluripotent Stem Cells

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1220-1220
Author(s):  
Hao Bai ◽  
Yin-Liang Xie ◽  
Yong-Xing Gao ◽  
Tao Cheng ◽  
Zack Z Wang
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Endothelial Cells ◽  
Pluripotent Stem Cells ◽  
Hematopoietic Cells ◽  
Human Pluripotent Stem Cells ◽  
Embryoid Bodies ◽  
Mesoderm Induction ◽  
Tgf Beta ◽  
Free System

Abstract Abstract 1220 Derived from mesoderm precursors, hemangioblasts are bipotential common progenitors of hematopoietic cells and endothelial cells. The regulatory events controlling human hemangioblast development are largely unknown. Our previous study demonstrated that CD34 progenitors from human embryonic stem cells (hESCs) contain a population of cells that give rise to hematopoietic cells and endothelial cells. In this study, we established a serum-free and feeder-free system to investigate the signals that direct differentiation of human pluripotent stem cells (hPSCs), including hESCs and induced-pluripotent stem cells (hiPSCs). The hemangioblast development in hPSCs was established in three major steps: i) the formation of uniformed embryoid bodies (EBs) in hanging-drops, ii) the development of multipotential mesoderm precursors in cultivation of EBs, and iii) the generation of CD34+CD31+VE-cadherin+ progenitors, which have hemangioblast potential to give rise to both hematopoietic and endothelial cells (via hemogenic endothelium). The hESC-derived hematopoietic cells expressed CD41, CD61, CD117, CD71, and CD235a. The endothelial cells were CD31+ and VE-cadherin+ cells. We found that inhibition of Rho-associated coiled-coil forming protein serine/threonine kinase (ROCK) by Y27632 was critical for hanging-drop EB formation. Transforming growth factor beta (TGF-beta) is a key regulator of hematopoiesis with potent inhibitory effects on progenitor and stem cell proliferation. The expression of TGF-beta was increased during hPSC differentiation. Inhibition of TGF-beta signaling by the TGF-beta inhibitor, SB431542, significantly decreased the efficiency of CD34+ progenitors, suggesting a crucial role of TGF-beta signaling in hPSC differentiation. To further investigate when TGF-beta signaling affects hPSC differentiation to CD34+ progenitors, we added TGF-beta inhibitor at different stages of hPSC differentiation. We found that TGF-beta signaling is required for the early mesoderm induction. Inhibition of TGFb signaling before hemangioblast commitment during PSC differentiation significantly decreases the frequency of hemangioblasts (CD34+/CD31+/VE-cadherin+ cells) and the expression of mesoderm markers, such as Brachyury, Tie1, EOMES, and MIXL1. On the other hand, inhibition of TGF-beta signaling after mesoderm induction increases the number of CD34+/CD31+/VE-cadherin+ cells. EdU cell proliferation assays indicated that the proliferation of CD34+ progenitors were not affected by SB431542, suggesting that TGF-beta signaling suppresses hemangioblast generation. Our study demonstrated that TGF-beta signaling has the double-edged effect on hPSC differentiation to hemangioblasts. Disclosures: No relevant conflicts of interest to declare.

An activated form of type I serine/threonine kinase receptor TARAM-A reveals a specific signalling pathway involved in fish head organiser formation

Development ◽  
1996 ◽  
Vol 122 (12) ◽  
pp. 3735-3743 ◽  
Author(s):  
A. Renucci ◽  
V. Lemarchandel ◽  
F. Rosa
Keyword(s):  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Axis Formation ◽  
Mesoderm Induction ◽  
Type I ◽  
Tgf Beta ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Anterior Dorsal ◽  
Dorsal Mesoderm

The role of Transforming Growth Factor beta (TGF-beta)-related molecules in axis formation and mesoderm patterning in vertebrates has been extensively documented, but the identity and mechanisms of action of the endogenous molecules remained uncertain. In this study, we isolate a novel serine/threonine kinase type I receptor, TARAM-A, expressed during early zebrafish embryogenesis first ubiquitously and then restricted to dorsal mesoderm during gastrulation. A constitutive form of the receptor is able to induce the most anterior dorsal mesoderm rapidly and to confer an anterior organizing activity. By contrast, the wild-type form is only able to induce a local expansion of the dorsal mesoderm. Thus an activated form of TARAM-A is sufficient to induce dorsoanterior structures and TARAM-A may be activated by dorsally localized signals. Our data suggest the existence in fish of a specific TGF-beta-related pathway for anterior dorsal mesoderm induction, possibly mediated by TARAM-A and activated at the late blastula stage by localized dorsal determinant.

DVR-4 (bone morphogenetic protein-4) as a posterior-ventralizing factor in Xenopus mesoderm induction

Development ◽  
1992 ◽  
Vol 115 (2) ◽  
pp. 639-647 ◽  
Author(s):  
C.M. Jones ◽  
K.M. Lyons ◽  
P.M. Lapan ◽  
C.V. Wright ◽  
B.L. Hogan
Keyword(s):  
Growth Factors ◽  
Bone Morphogenetic Protein ◽  
Ectopic Expression ◽  
Body Plan ◽  
Bone Morphogenetic Protein 4 ◽  
Mesoderm Induction ◽  
Tgf Beta ◽  
Signalling Molecules ◽  
Morphogenetic Protein ◽  
Peptide Growth Factors

Establishment of mesodermal tissues in the amphibian body involves a series of inductive interactions probably elicited by a variety of peptide growth factors. Results reported here suggest that mesodermal patterning involves an array of signalling molecules including DVR-4, a TGF-beta-like molecule. We show that ectopic expression of DVR-4 causes embryos to develop with an overall posterior and/or ventral character, and that DVR-4 induces ventral types of mesoderm in animal cap explants. Moreover, DVR-4 overrides the dorsalizing effects of activin. DVR-4 is therefore the first molecule reported both to induce posteroventral mesoderm and to counteract dorsalizing signals such as activin. Possible interactions between these molecules resulting in establishment of the embryonic body plan are discussed.

Synergistic principles of development: overlapping patterning systems in Xenopus mesoderm induction

Development ◽  
1992 ◽  
Vol 116 (1) ◽  
pp. 1-12 ◽  
Author(s):  
D. Kimelman ◽  
J. L. Christian ◽  
R. T. Moon
Keyword(s):  
Experimental Data ◽  
Gene Families ◽  
Mesoderm Induction ◽  
Tgf Beta ◽  
Signalling Molecules ◽  
Working Model ◽  
Xenopus Development ◽  
Anterior Posterior

The first inductive event in Xenopus development establishes the mesoderm at the equator of the developing embryo. As part of this process, the dorsal-ventral and anterior-posterior axes of the embryo are initially established. A number of signalling molecules which may play a role in mesodermal induction and patterning have been identified in the last several years, including members of the FGF, TGF-beta and Wnt gene families. A variety of experiments, using either purified factors or injection of RNA encoding these factors, have added to the wealth of classical embryogical experimental data collected over the last century. We have synthesized some recent results with the classical data to provide a framework for examining the process of mesoderm induction, and to formulate putative roles for some of the different factors. We incorporate these ideas into a working model of mesoderm induction that provides a basis for future experimental directions. Finally, we suggest that mesoderm induction may not be a discrete set of well separated events, but instead may be a process involving partially overlapping signals that produce the same pattern.

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