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At least 19 recordsLinked to original sources
PubMed1994

A truncated bone morphogenetic protein 4 receptor alters the fate of ventral mesoderm to dorsal mesoderm: roles of animal pole tissue in the development of ventral mesoderm.

The biological effects of endogenous bone morphogenetic protein 4 (BMP-4), a member of the transforming growth factor beta family, on embryonic development of Xenopus laevis were investigated by using a functionally defective mutant of the BMP-4 receptor (delta mTFR11), which blocks the BMP signaling pathway. Injection of delta mTFR11 RNA into either the animal pole area or ventral marginal cells at the two-cell stage induced a dorsal phenotype in the explant of ventral mesoderm with animal pole tissue from stage 10+ embryo, even though the normal fate of this explant is a "mesenchymal ball" containing blood cells. These explants with the dorsal phenotype contained muscle, neural tissue, eye capsule, and cement gland. Northern blot analysis showed an increase of cardiac alpha-actin mRNA and a decrease of T alpha-globin mRNA expression, providing further evidence of a conversion from ventral to dorsal phenotype. Although injection of delta mTFR11 RNA did not induce mesoderm in an animal cap culture, the same tissue injected with delta mTFR11 RNA can alter the differentiation fate of uninjected ventral mesodermal explant from ventral to dorsal type, suggesting specific interaction of animal pole tissue and prospective ventral mesoderm in vivo.

Animals↗
PubMed2000

Signals from trunk paraxial mesoderm induce pronephros formation in chick intermediate mesoderm.

We used Pax-2 mRNA expression and Lim 1/2 antibody staining as markers for the conversion of chick intermediate mesoderm (IM) to pronephric tissue and Lmx-1 mRNA expression as a marker for mesonephros. Pronephric markers were strongly expressed caudal to the fifth somite by stage 9. To determine whether the pronephros was induced by adjacent tissues and, if so, to identify the inducing tissues and the timing of induction, we microsurgically dissected one side of chick embryos developing in culture and then incubated them for up to 3 days. The undisturbed contralateral side served as a control. Most embryos cut parallel to the rostrocaudal axis between the trunk paraxial mesoderm and IM before stage 8 developed a pronephros on the control side only. Embryos manipulated after stage 9 developed pronephric structures on both sides, but the caudal pronephric extension was attenuated on the cut side. These results suggest that a medial signal is required for pronephric development and show that the signal is propagated in a rostral to caudal sequence. In manipulated embryos cultured for 3 days in ovo, the mesonephros as well as the pronephros failed to develop on the experimental side. In contrast, embryos cut between the notochord and the trunk paraxial mesoderm formed pronephric structures on both sides, regardless of the stage at which the operation was performed, indicating that the signal arises from the paraxial mesoderm (PM) and not from axial mesoderm. This cut also served as a control for cuts between the PM and the IM and showed that signaling itself was blocked in the former experiments, not the migration of pronephric or mesonephric precursor cells from the primitive streak. Additional control experiments ruled out the need for signals from lateral plate mesoderm, ectoderm, or endoderm. To determine whether the trunk paraxial mesoderm caudal to the fifth somite maintains its inductive capacity in the absence of contact with more rostral tissue, embryos were transected. Those transected below the prospective level of the fifth somite expressed Pax-2 in both the rostral and the caudal isolates, whereas embryos transected rostral to this level expressed Pax-2 in the caudal isolate only. Thus, a rostral signal is not required to establish the normal pattern of Pax-2 expression and pronephros formation. To determine whether paraxial mesoderm is sufficient for pronephros induction, stage 7 or earlier chick lateral plate mesoderm was cocultured with caudal stage 8 or 9 quail somites in collagen gels. Pax-2 was expressed in chick tissues in 21 of 25 embryos. Isochronic transplantation of stage 4 or 5 quail node into caudal chick primitive streak resulted in the generation of ectopic somites. These somites induced ectopic pronephroi in lateral plate mesoderm, and the IM that received signals from both native and ectopic somites formed enlarged pronephroi with increased Pax-2 expression. We conclude that signals from a localized region of the trunk paraxial mesoderm are both required and sufficient for the induction of the pronephros from the chick IM. Studies to identify the molecular nature of the induction are in progress.

Animals↗
PubMed1985

Dorsal lateral plate mesoderm influences proliferation and differentiation of hemopoietic stem cells derived from ventral lateral plate mesoderm during early development of Xenopus laevis embryos.

The developmental pattern of dorsal and ventral lateral plate mesoderm was examined by explanting these regions and culturing them in serum-free conditions for 10 days. Under these circumstances, ventral lateral plate mesoderm gave rise to hemopoietic cells, and dorsal lateral plate mesoderm gave rise to stromal cells. Comparison of the differentiation pattern observed when dorsal and ventral mesoderm were cultured separately versus the pattern observed when dorsal and ventral mesoderm were cultured together revealed that dorsal mesoderm influenced the differentiation of ventral-derived hemopoietic cells. Ventral mesoderm cultured alone gave rise to erythrocytes and granulocytes. However, in the presence of dorsal mesoderm, ventral mesoderm gave rise to erythrocytes, monocytes, and granulocytes. Moreover, dorsal mesoderm functioned to maintain the proliferation and differentiation potential of progenitor cells as determined using a colony-forming cell assay (CFU-c).

Animals↗
PubMed2000

Development of chick axial mesoderm: specification of prechordal mesoderm by anterior endoderm-derived TGFbeta family signalling.

Two populations of axial mesoderm cells can be recognised in the chick embryo, posterior notochord and anterior prechordal mesoderm. We have examined the cellular and molecular events that govern the specification of prechordal mesoderm. We report that notochord and prechordal mesoderm cells are intermingled and share expression of many markers as they initially extend out of Hensen's node. In vitro culture studies, together with in vivo grafting experiments, reveal that early extending axial mesoderm cells are labile and that their character may be defined subsequently through signals that derive from anterior endodermal tissues. Anterior endoderm elicits aspects of prechordal mesoderm identity in extending axial mesoderm by repressing notochord characteristics, briefly maintaining gsc expression and inducing BMP7 expression. Together these experiments suggest that, in vivo, signalling by anterior endoderm may determine the extent of prechordal mesoderm. The transforming growth factor (beta) (TGFbeta) superfamily members BMP2, BMP4, BMP7 and activin, all of which are transiently expressed in anterior endoderm mimic distinct aspects of its patterning actions. Together our results suggest that anterior endoderm-derived TGFbetas may specify prechordal mesoderm character in chick axial mesoderm.

Activins↗
PubMed1987

The organization of mesodermal pattern in Xenopus laevis: experiments using a Xenopus mesoderm-inducing factor.

In this paper, we study the mechanism by which a Xenopus cell line-derived mesoderm-inducing factor (MIF) might establish the spatial pattern of cellular differentiation in the mesoderm. The effects of the factor on competent animal pole tissue are consistent with it being identical to the natural mesoderm-inducing factor. The signal can only act on those membrane domains of the animal pole that face the blastocoel, but it can be stably recorded there, such that axial mesoderm is formed, after 15 min exposure or less. This exposure can end some hours, or several cell cycles, before the onset of RNA synthesis yet nevertheless be fully effective, although competence to respond also extends well after the onset of transcription. Exposure of the entire blastocoel lining of intact embryos to MIF causes a synchronous and sudden transformation of the behaviour and adhesive properties of all inner animal cap cells. This transformation mimics and is contemporaneous with the involution behaviour of normal mesoderm in the early gastrula marginal zone. Although high concentrations of MIF totally disorganize gastrulation, lower concentrations permit gastrulation to proceed. However, the pattern of mesoderm in these embryos is disrupted and ectopic mesoderm is formed around the blastocoel remnant. When MIF is injected directly into blastomeres, rather than into the blastocoel, it has no effect. This suggests that the molecule is secreted from source cells and affects target cells through an extracellular receptor. Finally, we show that small pieces of animal pole tissue recently exposed to MIF go on to produce morphogenetic signals perhaps distinct from MIF. We discuss the role of these signals in establishing and modifying the spatial pattern of cellular differentiation in the mesoderm of Xenopus.

Animals↗
PubMed1992

Brown adipose tissue as a derivative of mesoderm grafted below the kidney capsule. A model for differentiation of isolated rat mesoderm.

During development, mesoderm differentiates into connective tissue, cartilage, bone, muscle and kidney. In experimental conditions the developmental spectrum of mesoderm grafted below the kidney capsule is reduced so that mostly brown adipose tissue (BAT) appears. Since BAT is a particular tissue with a specific developmental pattern, the structural and functional characteristics of experimentally developed BAT were analyzed in the present study. Mesoderm from nine-day-old rat embryos was grafted below the kidney capsule of adult rats and one month later the BAT-containing tumors were analyzed. The ultrastructural and morphometrical data of BAT-containing tumors were the same as in BAT developed in situ. Tissue-specific mRNA for uncoupling protein (UCP) was expressed in BAT-containing tumors, and immuno-electron microscopical analysis showed that mitochondria of these brown adipocytes contained UCP. Injections of noradrenaline and exposure of BAT-tumor-bearing rats to cold stress increased both the amount of UCP and the expression of UCP mRNA in tumors of BAT; i.e., experimentally developed BAT entirely resembled standard BAT. It is proposed that mesoderm isolated and displaced below the kidney capsule lacks the inductive stimuli of ectoderm and endoderm, and as a result mesoderm can not realize the natural pattern of differentiation. Here, in a new environment, mesoderm is exposed to new stimuli which induce differentiation of mesoderm into BAT, probably through neuro-vascular elements from the medial side of the kidney (BAT area). Thus, although mesoderm contains a wide differentiation capacity, it can differentiate into only one type of tissue, depending on the presence and range of inductive stimuli.

Adipose Tissue, Brown↗
PubMed1997

Mixed cultures of avian blastoderm cells and the quail mesoderm cell line QCE-6 provide evidence for the pluripotentiality of early mesoderm.

During the early stages of embryogenesis, the mesoderm gives rise to cells of the cardiovascular system which include cardiac myocytes and vascular endothelial and red blood cells. We have investigated the development of these cell phenotypes using aggregate cultures of avian blastoderm cells, which replicated mesodermal cell diversification. The cell phenotypes expressed by the blastoderm cells were dependent upon the age of the blastoderm cells, with Hamburger-Hamilton stage 3 or 4 cells giving rise to endothelial and red blood cells and stage 5 cells producing endothelial and myocardial cells. To begin to understand the stage dependency of the cellular diversification of these aggregate cultures, we treated the cultures with various signaling factors that have been shown to be present in the early avian embryo. These experiments showed that stem cell factor and TGF alpha altered cell phenotypes by stimulating red blood cell and myocardial differentiation, respectively. The ability of these growth factors to shift the differentiation profile of aggregate cultures demonstrated the plasticity of early embryonic cells. To explore the diversification of individual mesodermal cells, labeled QCE-6 cells were incorporated within these blastoderm aggregate cultures. Previous studies have shown that this quail mesodermal cell line possesses characteristics of early nondifferentiated mesodermal cells and can be induced to express either myocardial or endothelial cell phenotypes (C. A. Eisenberg and D. M. Bader, 1996, Circ. Res. 78, 205-216). In the present study, we show that when these cells were cultured as a component of blastoderm cell aggregates, they differentiated into fully contractile cardiomyocytes or endothelial or red blood cells. Moreover, QCE-6 cell differentiation was in accordance with that displayed by the blastoderm cells. Specifically, QCE-6 cells differentiated into red blood cells when cultured within stage 3 or stage 4, but not stage 5, blastoderm cell aggregates. Accordingly, the differentiation of QCE-6 cells into beating cardiomyocytes only occurred when these cells were incorporated into stage 5 blastoderm cell aggregates. The identical sorting and differentiation patterns that were exhibited by QCE-6 and blastoderm cells suggest that expression of differentiated cell types within the early mesoderm is directed by the surrounding environment without immediate cellular commitment. In addition, these results provide further evidence that QCE-6 cells are representative of a multipotential mesodermal stem cell and that they possess the potential to exhibit fully differentiated cell phenotypes.

Animals↗
PubMed2002

Mesodermal and axial determinants contribute to mesoderm regionalization in Bufo arenarum embryos.

The existence of mesodermal determinants in the equator of Bufo arenarum embryos has been previously demonstrated. In this work, their role in dorso-ventral regionalization of mesoderm was studied by transferring the determinants to animal blastomeres. The transfer was performed by cleavage reorientation and cytoplasmic microinjection. Forced inclination during early cleavage caused deviation of the third cleavage plane and annexation of equatorial cytoplasm into animal quartets. Animal blastomeres from embryos oriented with the dorsal side up, incorporated ventro-equatorial cytoplasm and formed blood cells, mesenchyme, and coelomic epithelium. In contrast, animal blastomeres from embryos oriented with the ventral side up, acquired dorso-equatorial cytoplasm and developed notochord, somites, mesenchyme, coelomic epithelium and nervous tissue. In order to investigate if this dorso-ventral differentiation pattern responds to an interaction of mesodermal and axial factors, isolated 8-cell-stage animal quartets were microinjected with subcortical cytoplasm from: (a) the ventro-equatorial region of synchronous embryos; (b) the vegetal pole of uncleaved eggs; (c) a combination of both cytoplasms. As expected, the implanted ventro-equatorial cytoplasm promoted ventral mesoderm differentiation. Conversely, the joint transfer of ventro-equatorial cytoplasm and vegetal pole cytoplasm behaved as the dorso-equatorial cytoplasm, promoting dorso-lateral mesoderm and neural formation. Thus, mesoderm regionalization in B. arenarum embryos seems to be caused by a concurrent action of both mesodermal and axial determinants.

Animals↗
PubMed1998

Mesoderm induction by heterodimeric AP-1 (c-Jun and c-Fos) and its involvement in mesoderm formation through the embryonic fibroblast growth factor/Xbra autocatalytic loop during the early development of Xenopus embryos.

We have previously demonstrated the involvement of AP-1/Jun in fibroblast growth factor (FGF) signaling by loss-of-function assay (Dong, Z., Xu, R.-H., Kim, J., Zhan, S.-N., Ma, W.-Y., Colburn, N. H., and Kung, H. (1996) J. Biol. Chem. 271, 9942-9946). Further investigations by gain-of-function are reported in this study. AP-1 transactivation activity was increased by the treatment of animal cap explants with FGF. Ectopic overexpression of two components of AP-1 (c-jun and c-fos together, but not alone) produced posteriorized embryos and induced mesoderm formation in animal cap explants, indicating that both AP-1 heterodimers are required for mesoderm induction. Since Ras/AP-1 functions downstream of FGF signaling, we then tested the involvement of Ras/AP-1 in mesoderm maintenance mediated by embryonic FGF/Xbra using dominant-negative mutants. Mesoderm maintenance mediated by embryonic FGF/Xbra was blocked by dominant-negative mutants of Ras/AP-1, and AP-1 enhanced the expression of Xbra. Further studies demonstrated the inhibition of Ras/AP-1-mediated mesoderm formation by dominant-negative mutants of the FGF receptor and Xbra. These results indicate that Ras/AP-1 and FGF/Xbra signals are involved in the mesoderm maintenance machinery and mesoderm formation through the synergistic action of the diversified signal pathways derived from the FGF/Xbra autocatalytic loop.

Animals↗
PubMed1991

Xwnt-8, a Xenopus Wnt-1/int-1-related gene responsive to mesoderm-inducing growth factors, may play a role in ventral mesodermal patterning during embryogenesis.

In amphibian embryos, formation of the basic body plan depends on positional differences in the mesoderm. Although peptide growth factors involved in mesoderm induction have tentatively been identified, additional signals are required to generate pattern in this tissue. We have isolated a Xenopus cDNA for a Wnt-1 related gene, designated Xwnt-8, which is activated in response to mesoderm-inducing growth factors. Xwnt-8 transcripts are transiently expressed, being most abundant during gastrulation at which time expression is confined primarily to ventral mesodermal cells. Embryos dorsoanteriorized by exposure to lithium exhibit greatly reduced levels of Xwnt-8 mRNA, supporting a correlation between Xwnt-8 expression and a ventral mesodermal cell fate. Surprisingly, ectopic expression of Xwnt-8 in embryos causes a dorsoanterior-enhanced phenotype. These findings suggest that Xwnt-8 may be a secondary signalling agent which is produced in response to mesoderm-inducing factors and is involved in the early steps of mesodermal patterning.

Animals↗
PubMed1992

Expression of tenascin mRNA in mesoderm during Xenopus laevis embryogenesis: the potential role of mesoderm patterning in tenascin regionalization.

In Xenopus embryos, the extracellular matrix (ECM) protein tenascin (TN) is expressed dorsally in a very restricted pattern. We have studied the spatial and temporal expression of TN mRNA in tailbud-stage embryos by RNAase protection and in situ hybridization using a cDNA probe for Xenopus TN obtained by PCR amplification. We report that TN transcripts are principally expressed in cells dispersed around the neural tube and notochord as well as in myotome and sclerotome cells. No TN mRNA could be detected in lateral plate mesoderm, but expression was detectable beneath tail fin epidermis. In a second series of experiments, we studied the expression of TN mRNA and protein in combinations between animal and vegetal stage-6 blastomeres and in stage-8 blastula animal caps treated with activin A or basic fibroblastic growth factor (b-FGF). Isolated animal cap tissue cultured alone differentiates into epidermis, which expresses neither TN protein nor TN mRNA. TN expression is, however, elicited in response to isolated dorsal vegetal blastomeres and in response to high concentrations of activin, both of which treatments lead to formation of muscle and/or notochord. Low concentrations of activin, and ventral vegetal blastomeres, treatments that induce mesoderm of ventral character, are poor inducers of TN. However, b-FGF, which also induces ventral mesoderm, elicits strong expression. These results indicate that TN regionalization is a complex process, dependent both on the pattern of differentiation of mesodermal tissues and on the agent with which they are induced. The data further show that "ventral mesoderm" induced by low concentrations of activin is distinct from that induced by b-FGF, and imply that activin induces ventral mesoderm of the trunk while b-FGF induces posterior mesoderm of the tailbud.

Animals↗
PubMed1988

Mesoderm induction in Xenopus laevis: responding cells must be in contact for mesoderm formation but suppression of epidermal differentiation can occur in single cells.

When Xenopus embryos are cultured in calcium- and magnesium-free medium (CMFM), the blastomeres lose adhesion but continue dividing to form a loose heap of cells. If divalent cations are restored at the early gastrula stage the cells re-adhere and eventually form muscle (a mesodermal cell type) as well as epidermis. If, however, the cells are dispersed during culture in CMFM, muscle does not form following reaggregation although epidermis does. This suggests that culturing blastomeres in a heap allows the transmission of mesoderm-induction signals from cell to cell while dispersion effectively dilutes the signal. In this paper, we have attempted to substitute for cell proximity by culturing dispersed blastomeres in XTC mesoderm-inducing factor (MIF). We find that dispersed cells do not respond to XTC-MIF by forming mesodermal cell types after reaggregation, but the factor does inhibit epidermal differentiation. One interpretation of this observation is that an early stage in mesoderm induction is the suppression of epidermal differentiation and that formation of mesoderm may require contact-mediated signals that are produced in response to XTC-MIF. We have gone on to study the suppression of epidermal differentiation in more detail. We find that this is a dose-dependent phenomenon that can occur in single cells in the absence of cell division. Animal pole blastomeres become more difficult to divert from epidermal differentiation at later stages of development and by stage 12 they are 'determined' to this fate. Fibroblast growth factor (FGF) also suppresses epidermal differentiation in isolated animal pole blastomeres and transforming growth factor-beta 1 acts synergistically with FGF in doing so.

Animals↗
PubMed1997

Wnt-11 is expressed in early avian mesoderm and required for the differentiation of the quail mesoderm cell line QCE-6.

The beginning of mesodermal development involves the aggregation of newly gastrulated cells into epithelial fields, as a prelude to organ formation. To analyze the molecular regulation of this initial patterning, we have focused on the Wnt family of secreted signaling proteins, molecules which have been shown to promote embryonic patterning by regulating cell-cell associations. In this study, we show that the Wnt-11 gene is expressed by newly gastrulated mesoderm cells within avian embryos. The expression pattern of Wnt-11 also suggests that it may be involved in formation of the cardiogenic fields and somites. Subsequently, we utilized the quail mesoderm cell line QCE-6 as a culture model for examining the influence of Wnt-11 on early mesoderm cell differentiation. This cell line has been shown to be representative of early nondifferentiated mesoderm cells and has the potential to differentiate into cardiomyocytes, endothelial or red blood cells. Similar to early mesoderm cells, QCE-6 cells express Wnt-11. We have engineered stable transfectants of these cells that produce either diminished or enhanced levels of Wnt-11 protein. Our studies show that Wnt-11 regulates cellular interactions of QCE-6 cells, as demonstrated by alterations in contact-inhibited growth, tight and gap junction formation and plakoglobin expression. Both the morphology and growth factor-induced differentiation of QCE-6 cells are regulated in a cooperative fashion by Wnt-11 and fibronectin. These results, described in detail below, demonstrate the uniqueness of QCE-6 cells as a culture system for analyzing Wnt activity. In particular, QCE-6 cells are the first cell line that has demonstrated: (1) Wnt-dependent differentiation; (2) concentration-variable responses to Wnt protein; and (3) altered cell phenotypes as a direct response to Wnt-5a class proteins (e.g. Wnt-4 and Wnt-11).

Amino Acid Sequence↗
PubMed1997

A vegetally localized T-box transcription factor in Xenopus eggs specifies mesoderm and endoderm and is essential for embryonic mesoderm formation.

Pattern formation in early embryogenesis is guided by maternal, localized determinants and by inductive interactions between cells. In Xenopus eggs, localized molecules have been identified and some, such as Vg1 and Xwnt-11, can specify cell fates by functioning as inducers or patterning agents. We have used differential screening to identify new Xenopus genes that regulate mesodermal patterning, and we have isolated a new member of the T-box family of transcription factors. This gene, named Brat, is expressed maternally and its transcripts are localized to the vegetal hemisphere of the egg. During early embryonic cleavage, Brat mRNA becomes partitioned primarily within vegetal cells that are fated to form the endoderm. Zygotic expression of Brat begins at the onset of gastrulation within the presumptive mesoderm of the marginal zone. Consistent with its zygotic expression pattern, Brat induces, in a dose-dependent manner, a full spectrum of mesodermal genes that mark tissues across the dorsal-ventral axis, from the blood through the Spemann organizer. Brat also induces endoderm, consistent with its vegetal localization, making Brat a good candidate for a maternal determinant of the endoderm. We tested whether endogenous Brat is required for mesoderm formation by expressing a dominant-negative, transcriptional repressor form of Brat in embryos. This treatment inhibited mesoderm formation and severely disrupted normal development, thereby establishing that Brat plays a critical role in embryonic mesoderm formation and body patterning.

Activins↗
PubMed1989

Mesoderm induction by the mesoderm of Xenopus neurulae.

Combinations were made between explants of mesoderm from the archenteron roof of early Xenopus neurulae and explants of ectoderm from mid-blastulae. In each combination one component was labeled with the fluorescent lineage label RDA (rhodamine-dextran-amine). Frequent and large mesoderm inductions, consisting mainly of muscle, were found where the presomite plate was used as the inducer. Less frequent and smaller mesoderm inductions were found when notochord was used as the inducer. We conclude that induced mesoderm can itself be active as a mesoderm inducing tissue. If this capability is acquired in the blastula then it follows that mesoderm induction must propagate from cell to cell and its spread be antagonized by some other factor.

Animals↗
PubMed2000

The VT+ and VT- isoforms of the fibroblast growth factor receptor type 1 are differentially expressed in the presumptive mesoderm of Xenopus embryos and differ in their ability to mediate mesoderm formation.

Previously, we cloned a variant form of the type 1 fibroblast growth factor receptor (FGFR1), FGFR-VT-, from Xenopus embryos (Gillespie, L. L., Chen, G., and Paterno, G. D. (1995) J. Biol. Chem. 270, 22758-22763). This isoform differed from the reported FGFR1 sequence (FGFR-VT+) by a 2-amino acid deletion, Val(423)-Thr(424), in the juxtamembrane region. This deletion arises from the use of an alternate 5' splice donor site, and the activity of the VT+ and VT- forms of the FGFR1 was regulated by phosphorylation at this site. We have now investigated the expression pattern and function of these two isoforms in mesoderm formation in Xenopus embryos. Cells within the marginal zone are induced to form mesoderm during blastula stages. RNase protection analysis of blastula stage embryos revealed that the VT+ isoform was expressed throughout the embryo but that the VT- isoform was expressed almost exclusively in the marginal zone. The ratio of VT+:VT- transcripts in the marginal zone indicated that the VT+ form was predominant throughout blastula stages except for a brief interval, coinciding with the start of zygotic transcription, when a dramatic increase in VT- expression levels was detected. This increase could be mimicked in part by treatment of animal cap explants with FGF-2. Overexpression of the VT+ isoform in Xenopus embryos resulted in development of tadpoles with severe reductions in trunk and tail structures, while embryos overexpressing the VT- isoform developed normally. A standard mesoderm induction assay revealed that a 10-fold higher concentration of FGF-2 was required to reach 50% induction in VT+-overexpressing animal cap explants compared with those overexpressing the VT- isoform. Furthermore, little or no expression of the panmesodermal marker Brachyury (Xbra) was detected in VT+-overexpressing embryos, while VT--overexpressing embryos showed normal staining. This demonstrates that VT+ overexpression had a negative effect on mesoderm formation in vivo. These data are consistent with a model in which mesoderm formation in vivo is regulated, at least in part, by the relative expression levels of the VT+ and VT- isoforms.

Animals↗
PubMed1995

Nodal-related signals induce axial mesoderm and dorsalize mesoderm during gastrulation.

Mouse embryos homozygous for a null mutation in nodal arrest development at early gastrulation and contain little or no embryonic mesoderm. Here, two Xenopus nodal-related genes (Xnr-1 and Xnr-2) are identified and shown to be expressed transiently during embryogenesis, first within the vegetal region of late blastulae and later in the marginal zone during gastrulation, with enrichment in the dorsal lip. Xnrs and mouse nodal function as dose-dependent dorsoanterior and ventral mesoderm inducers in whole embryos and explanted animal caps. Using a plasmid vector to produce Xnr proteins during gastrulation, we show that, in contrast to activin and other TGF beta-like molecules, Xnr-1 and Xnr-2 can dorsalize ventral marginal zone explants and induce muscle differentiation. Xnr signalling also rescues a complete embryonic axis in UV-ventralized embryos. The patterns of Xnr expression, the activities of the proteins and the phenotype of mouse nodal mutants, all argue strongly that a signaling pathway involving nodal, or nodal-related peptides, is an essential conserved element in mesoderm differentiation associated with vertebrate gastrulation and axial patterning.

Amino Acid Sequence↗
PubMed1996

Inhibition of Xbra transcription activation causes defects in mesodermal patterning and reveals autoregulation of Xbra in dorsal mesoderm.

The Brachyury (T) gene is required for formation of posterior mesoderm and for axial development in both mouse and zebrafish embryos. In this paper, we first show that the Xenopus homologue of Brachyury, Xbra, and the zebrafish homologue, no tail (ntl), both function as transcription activators. The activation domains of both proteins map to their carboxy terminal regions, and we note that the activation domain is absent in two zebrafish Brachyury mutations, suggesting that it is required for gene function. A dominant-interfering Xbra construct was generated by replacing the activation domain of Xbra with the repressor domain of the Drosophila engrailed protein. Microinjection of RNA encoding this fusion protein allowed us to generate Xenopus and zebrafish embryos which show striking similarities to genetic mutants in mouse and fish. These results indicate that the function of Brachyury during vertebrate gastrulation is to activate transcription of mesoderm-specific genes. Additional experiments show that Xbra transcription activation is required for regulation of Xbra itself in dorsal, but not ventral, mesoderm. The approach described in this paper, in which the DNA-binding domain of a transcription activator is fused to the engrailed repressor domain, should assist in the analysis of other Xenopus and zebrafish transcription factors.

Animals↗