Blastomere derivation and domains of gene expression in the Spemann Organizer of Xenopus laevis

Development ◽  
1995 ◽  
Vol 121 (11) ◽  
pp. 3505-3518 ◽  
Author(s):  
M.A. Vodicka ◽  
J.C. Gerhart
Keyword(s):  
Gene Expression ◽  
Marginal Zone ◽  
Early Stage ◽  
Fate Map ◽  
Early Cleavage ◽  
Spemann Organizer ◽  
Spemann's Organizer ◽  
Cell Embryo ◽  
Early Gastrula

Spemann's Organizer, located in the dorsal marginal zone of the amphibian gastrula, induces and differentiates dorsal axial structures characteristic of this and other vertebrates. To trace the cellular origins of the Xenopus Organizer, we labelled dorsal blastomeres of three of the four tiers (A, B and C) of the 32-cell embryo with green, red and blue fluorescent lineage tracers. A strong vegetalward displacement of labelled clones occurs between the late blastula and early gastrula stages but clones mix only slightly at their borders. The typical early gastrula Organizer is composed of approximately 10% A1 progeny in its animalmost region, 70% B1 progeny in the central region, and 20% C1 progeny in vegetal and deep regions. Variability in the composition of the early gastrula Organizer results from variability in the position of early cleavage planes and in pregastrulation movements. As the Organizer involutes during gastrulation, forming dorsal axial mesoderm, clonal boundaries are greatly dispersed by cell intermixing. Within a clone, deep cells are displaced and intermixed more than superficial cells. Variability in the distribution of progeny in the dorsal axial mesoderm of the late gastrula results mostly from variable intermixing of cells during gastrulation. Experiments to perturb later developmental events by molecular or embryonic manipulations at an early stage must take this variability into account along with the majority distributions of the fate map. Within the early gastrula Organizer, the genes Xbra, goosecoid, noggin and xNR3 are expressed differently in the animal-vegetal and superficial-deep dimensions. In situ hybridization and lineage labelling define distinct regions of the dorsal marginal zone. By the end of gastrulation, dorsal axial mesoderm cells derived from the Organizer have altered their expression of the genes Xbra, goosecoid, noggin and xNR3. At a given stage, a cell's position in the embryo rather than its lineage may be more important in determining which genes it will express.

XIPOU 2 is a potential regulator of Spemann's Organizer

Development ◽  
1997 ◽  
Vol 124 (6) ◽  
pp. 1179-1189 ◽  
Author(s):  
S.E. Witta ◽  
S.M. Sato
Keyword(s):  
Gene Expression ◽  
Wnt Pathway ◽  
Marginal Zone ◽  
Branchial Arch ◽  
Class Iii ◽  
Domain Family ◽  
Potent Inducer ◽  
Pou Domain ◽  
Spemann's Organizer ◽  
Dorsal Mesoderm

XIPOU 2, a member of the class III POU-domain family, is expressed initially at mid-blastula transition (MBT) and during gastrulation in the entire marginal zone mesoderm, including Spemann's Organizer (the Organizer). To identify potential targets of XIPOU 2, the interaction of XIPOU 2 with other genes co-expressed in the Organizer was examined by microinjecting XIPOU 2's mRNA into the lineage of cells that contributes to the Organizer, head mesenchyme and prechordal plate. XIPOU 2 suppresses the expression of a number of dorsal mesoderm-specific genes, including gsc, Xlim-1, Xotx2, noggin and chordin, but not Xnot. As a consequence of the suppression of dorsal mesoderm gene expression, bone morphogenetic factor-4 (Bmp-4), a potent inducer of ventral mesoderm, is activated in the Organizer. Gsc is a potential target of XIPOU 2. XIPOU 2 is capable of binding a class III POU protein binding site (CATTAAT) that is located within the gsc promoter, in the activin-inducible (distal) element. Furthermore, XIPOU 2 suppresses the activation of the gsc promoter by activin signaling. At the neurula and tailbud stages, dorsoanterior structures are affected: embryos displayed micropthalmia and the loss of the first branchial arch, as detected by the expression of pax-6, Xotx2 and en-2. By examining events downstream from the Wnt and chordin pathways, we determined that XIPOU 2, when overexpressed, acts specifically in the Organizer, downstream from GSK-3beta of the Wnt pathway and upstream from chordin. The interference in dorsalizing events caused by XIPOU 2 was rescued by chordin. Thus, in addition to its direct neuralizing ability, in a different context, XIPOU 2 has the potential to antagonize dorsalizing events in the Organizer.

scholarly journals The epithelium of the dorsal marginal zone of Xenopus has organizer properties

Development ◽  
1992 ◽  
Vol 116 (4) ◽  
pp. 887-899 ◽  
Author(s):  
J. Shih ◽  
R. Keller
Keyword(s):  
Neural Tube ◽  
Marginal Zone ◽  
Tissue Differentiation ◽  
Host Cells ◽  
Mesoderm Induction ◽  
Epithelial Layer ◽  
Spemann Organizer ◽  
Deep Region ◽  
Early Gastrula ◽  
Axial Development

We have investigated the properties of the epithelial layer of the dorsal marginal zone (DMZ) of the Xenopus laevis early gastrula and found that it has inductive properties similar to those of the entire Spemann organizer. When grafts of the epithelial layer of the DMZ of early gastrulae labelled with fluorescein dextran were transplanted to the ventral sides of unlabelled host embryos, they induced secondary axes composed of notochord, somites and posterior neural tube. The organizer epithelium rescued embryos ventralized by UV irradiation, inducing notochord, somites and posterior neural tube in these embryos, while over 90% of ventralized controls showed no such structures. Combinations of organizer epithelium and ventral marginal zone (VMZ) in explants of the early gastrula resulted in convergence, extension and differentiation of dorsal mesodermal tissues, whereas similar recombinants of nonorganizer epithelium and the VMZ did none of these things. In all cases, the axial structures forming in response to epithelial grafts were composed of labelled graft and unlabelled host cells, indicating an induction by the organizer epithelium of dorsal, axial morphogenesis and tissue differentiation among mesodermal cells that otherwise showed non-axial development. Serial sectioning and scanning electron microscopy of control grafts shows that the epithelial organizer effect occurs in the absence of contaminating deep cells adhering to the epithelial grafts. However, labelled organizer epithelium grafted to the superficial cell layer contributed cells to deep mesodermal tissues, and organizer epithelium developed into mesodermal tissues when deliberately grafted into the deep region. This shows that these prospective endodermal epithelial cells are able to contribute to mesodermal, mesenchymal tissues when they move or are moved into the deep environment. These results suggest that in normal development, the endodermal epithelium may influence some aspects of the cell motility underlying the mediolateral intercalation (see Shih, J. and Keller, R. (1992) Development 116, 901–914), as well as the tissue differentiation of mesodermal cells. These results have implications for the analysis of mesoderm induction and for analysis of variations in the differentiation and morphogenetic function of the marginal zone in different species of amphibians.

A fate map of superficial and deep circumblastoporal cells in the early gastrula of Pleurodeles waltl

Development ◽  
1992 ◽  
Vol 114 (1) ◽  
pp. 135-146 ◽  
Author(s):  
M. Delarue ◽  
S. Sanchez ◽  
K.E. Johnson ◽  
T. Darribere ◽  
J.C. Boucaut
Keyword(s):  
Marginal Zone ◽  
Cell Lineage ◽  
Tail Bud ◽  
Fate Map ◽  
Pleurodeles Waltl ◽  
Early Gastrula

We have determined the fate of presumptive mesodermal cells in the early Pleurodeles waltl gastrula. We labeled all cells in a gastrula with RLDx cell lineage tracer and superficial cells with 125I and then grafted small pieces of the marginal zone orthotopically into unlabeled host embryos. Labeled progeny were identified in sectioned embryos at the tail bud stage. The use of double-labeled grafts allowed us to study the relative contributions by superficial and deep cells to different derivatives. We found that the presumptive regions are generally distributed according to classical fate maps for urodeles but that the boundaries between presumptive regions are indistinct, due to extensive intermingling between cells at the edges of grafted regions. We have shown that there is a high dorsal to low ventral gradient of mixing between superficial and deep cells.

Bmp-4 acts as a morphogen in dorsoventral mesoderm patterning in Xenopus

Development ◽  
1997 ◽  
Vol 124 (12) ◽  
pp. 2325-2334 ◽  
Author(s):  
R. Dosch ◽  
V. Gawantka ◽  
H. Delius ◽  
C. Blumenstock ◽  
C. Niehrs
Keyword(s):  
Gene Expression ◽  
Long Range ◽  
Marginal Zone ◽  
Cell Types ◽  
Dorsoventral Patterning ◽  
Mesodermal Cell ◽  
Amphibian Embryos ◽  
Mesoderm Patterning ◽  
Early Gastrula ◽  
Different Doses

The marginal zone is a ring of tissue that gives rise to a characteristic dorsoventral pattern of mesoderm in amphibian embryos. Bmp-4 is thought to play an important role in specifying ventral mesodermal fate. Here we show (1) that different doses of Bmp-4 are sufficient to pattern four distinct mesodermal cell types and to pattern gene expression in the early gastrula marginal zone into three domains, (2) that there is a graded requirement for a Bmp signal in mesodermal patterning, and (3) that Bmp-4 has long-range activity which can become graded in the marginal zone by the antagonizing action of noggin. The results argue that Bmp-4 acts as a morphogen in dorsoventral patterning of mesoderm.

The homeobox-containing gene XANF-1 may control development of the Spemann organizer

Development ◽  
1995 ◽  
Vol 121 (11) ◽  
pp. 3839-3847 ◽  
Author(s):  
A.G. Zaraisky ◽  
V. Ecochard ◽  
O.V. Kazanskaya ◽  
S.A. Lukyanov ◽  
I.V. Fesenko ◽  
...  
Keyword(s):  
Marginal Zone ◽  
Anterior Part ◽  
Local Maximum ◽  
Cell Stage ◽  
Spemann Organizer ◽  
Secondary Axis ◽  
Stage Embryo ◽  
Migration Of Cells ◽  
Early Gastrula ◽  
Xenopus Laevis Embryos

At the beginning of gastrulation the homeobox-containing gene, XANF-1, is expressed at a low level throughout the animal hemisphere of Xenopus laevis embryos, with a local maximum of expression in the region of the dorsal blastopore lip. By the end of gastrulation expression ceases everywhere except in the most anterior part of the neurectoderm. We have investigated the functions of this gene by microinjecting XANF-1 mRNA in the blastomeres of the 32-cell stage embryo and have observed the following effects. First, microinjections of the mRNA in the animal blastomeres and the blastomeres of the marginal zone elicited massive migration of cells to the interior of the embryo at the early gastrula stage. Second, overexpression of XANF-1 in the ventral marginal zone (VMZ) resulted in the appearance of an additional centre of gastrulation movements and the formation of a secondary axis. In addition we showed that synthetic XANF-1 mRNA was able to cause dorsal-type differentiation in VMZ explants extirpated from the microinjected embryos at the beginning of gastrulation. These results suggest that XANF-1 may control the main functions of cells of the Spemann organizer.

dino and mercedes, two genes regulating dorsal development in the zebrafish embryo

Development ◽  
1996 ◽  
Vol 123 (1) ◽  
pp. 95-102 ◽  
Author(s):  
M. Hammerschmidt ◽  
F. Pelegri ◽  
M.C. Mullins ◽  
D.A. Kane ◽  
F.J. van Eeden ◽  
...  
Keyword(s):  
Zebrafish Embryo ◽  
Marginal Zone ◽  
Expression Patterns ◽  
Body Plan ◽  
Anterior Region ◽  
Marker Genes ◽  
Amphibian Embryo ◽  
Altered Expression ◽  
Spemann Organizer ◽  
Early Gastrula

We describe two genes, dino and mercedes, which are required for the organization of the zebrafish body plan. In dino mutant embryos, the tail is enlarged at the expense of the head and the anterior region of the trunk. The altered expression patterns of various marker genes reveal that, with the exception of the dorsal most marginal zone, all regions of the early dino mutant embryo acquire more ventral fates. These alterations are already apparent before the onset of gastrulation. mercedes mutant embryos show a similar but weaker phenotype, suggesting a role in the same patterning processes. The phenotypes suggests that dino and mercedes are required for the establishment of dorsal fates in both the marginal and the animal zone of the early gastrula embryo. Their function in the patterning of the ventrolateral mesoderm and the induction of the neuroectoderm is similar to the function of the Spemann organizer in the amphibian embryo.

Plasticity of transposed rhombomeres: Hox gene induction is correlated with phenotypic modifications

Development ◽  
1995 ◽  
Vol 121 (9) ◽  
pp. 2707-2721 ◽  
Author(s):  
A. Grapin-Botton ◽  
M.A. Bonnin ◽  
L.A. McNaughton ◽  
R. Krumlauf ◽  
N.M. Le Douarin
Keyword(s):  
Gene Expression ◽  
Neural Tube ◽  
Hox Genes ◽  
Gene Expression Pattern ◽  
Fate Map ◽  
Hox Gene ◽  
Inductive Signal ◽  
New Location ◽  
Chick And Quail Embryos

In this study we have analysed the expression of Hoxb-4, Hoxb-1, Hoxa-3, Hoxb-3, Hoxa-4 and Hoxd-4 in the neural tube of chick and quail embryos after rhombomere (r) heterotopic transplantations within the rhombencephalic area. Grafting experiments were carried out at the 5-somite stage, i.e. before rhombomere boundaries are visible. They were preceeded by the establishment of the precise fate map of the rhombencephalon in order to determine the presumptive territory corresponding to each rhombomere. When a rhombomere is transplanted from a caudal to a more rostral position it expresses the same set of Hox genes as in situ. By contrast in many cases, if rhombomeres are transplanted from rostral to caudal their Hox gene expression pattern is modified. They express genes normally activated at the new location of the explant, as evidenced by unilateral grafting. This induction occurs whether transplantation is carried out before or after rhombomere boundary formation. Moreover, the fate of the cells of caudally transplanted rhombomeres is modified: the rhombencephalic nuclei in the graft develop according to the new location as shown for an r5/6 to r8 transplantation. Transplantation of 5 consecutive rhombomeres (i.e. r2 to r6), to the r8 level leads to the induction of Hoxb-4 in the two posteriormost rhombomeres but not in r2,3,4. Transplantations to more caudal regions (posterior to somite 3) result in some cases in the induction of Hoxb-4 in the whole transplant. Neither the mesoderm lateral to the graft nor the notochord is responsible for the induction. Thus, the inductive signal emanates from the neural tube itself, suggesting that planar signalling and predominance of posterior properties are involved in the patterning of the neural primordium.

Identification of a transforming MYB-GATA1 fusion gene in acute basophilic leukemia: a new entity in male infants

Blood ◽  
2011 ◽  
Vol 117 (21) ◽  
pp. 5719-5722 ◽  
Author(s):  
Cathy Quelen ◽  
Eric Lippert ◽  
Stephanie Struski ◽  
Cécile Demur ◽  
Gwendoline Soler ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fusion Gene ◽  
Early Stage ◽  
Chromosomal Translocation ◽  
Rapid Amplification ◽  
First Time ◽  
Basophilic Leukemia ◽  
Infant Leukemia

Abstract Acute basophilic leukemia (ABL) is a rare subtype of acute leukemia with clinical features and symptoms related to hyperhistaminemia because of excessive growth of basophils. No known recurrent cytogenetic abnormality is associated with this leukemia. Rare cases of t(X;6)(p11;q23) translocation have been described but these were sporadic. We report here 4 cases of ABL with a t(X;6)(p11;q23) translocation occurring in male infants. Because of its location on chromosome 6q23, MYB was a good candidate gene. Our molecular investigations, based on fluorescence in situ hybridization and rapid amplification of cDNA ends, revealed that the translocation generated a MYB-GATA1 fusion gene. Expression of MYB-GATA1 in mouse lineage-negative cells committed them to the granulocyte lineage and blocked at an early stage of differentiation. Taken together, these results establish, for the first time, a link between a recurrent chromosomal translocation and the development of this particular subtype of infant leukemia.

Normal fates and states of specification of different regions in the axolotl gastrula

Development ◽  
1985 ◽  
Vol 86 (1) ◽  
pp. 247-269
Author(s):  
J. Herman Cleine ◽  
Jonathan M. W. Slack
Keyword(s):  
Marginal Zone ◽  
Culture Conditions ◽  
Early Embryo ◽  
Ambystoma Mexicanum ◽  
Specific Marker ◽  
Fate Map ◽  
Animal Pole ◽  
Glycoprotein Synthesis ◽  
Early Gastrula

A fate map was constructed for four regions of the early gastrula of Ambystoma mexicanum using orthotopic grafts from donors labelled with FLDx (fluoresceinated-lysinated-dextran). The region around the animal pole gave rise to epidermis only and did not include prospective neural plate. The dorsal marginal zone contributed to cephalic endoderm and to the whole length of the axial mesoderm (notochord and somites), the lateral marginal zone to lateroventral and somitic mesoderm, and the ventral marginal zone to lateroventral mesoderm. It was found that the dorsal marginal zone contributed relatively more to the anterior regions of the mesodermal mantle and the ventral marginal zone more to its posterior parts. The same regions of the gastrula and also vegetal yolky tissue were cultured as explants and labelled with tritiated mannose. Their glycoprotein synthesis pattern was compared to those of the neurula tissues to which they contribute in vivo. Animal pole explants synthesized large amounts of the epidermis-specific marker epimucin. Dorsal marginal zone explants did not synthesize epimucin but did make amounts of S2 and S6 indicative of mesoderm, as well as the notochord-specific markers S2·2 and S3·2. Lateral marginal zone explants showed the same pattern as the dorsal marginal zone including the two notochord-specific markers, although they do not contribute to notochord in vivo. Ventral marginal zone explants were more variable in their behaviour. Yolky tissue from the vegetal hemisphere of the gastrula or the archenteron floor of the neurula synthesized mainly polydisperse material of high molecular weight rather than discrete glycoproteins. The results indicate that at the early gastrula stage states of specification exist which correspond to the three germ layers, ecto-, meso- and endoderm. The ectodermal specification of animal pole explants is quite robust and cannot easily be changed by variation of the culture conditions. However treatment with a concentrated pellet of vegetalizing factor does induce a change to mesodermal specification, which is clearly detectable in the pattern of glycoprotein synthesis. Similar inductive interactions between different regions of the early embryo are thought to occur during normal development.

Molecular and Microscopic Analysis of Altered Gene Expression in Transgenic and Gene Targeted Mice

1996 ◽  
Vol 54 ◽  
pp. 12-13
Author(s):  
W. K. Jones ◽  
J. Robbins
Keyword(s):  
Gene Expression ◽  
Pulmonary Veins ◽  
Microscopic Analysis ◽  
Mouse Tissue ◽  
Adult Heart ◽  
Heavy Chains ◽  
Altered Gene Expression ◽  
Altered Gene ◽  
Pulmonary Myocardium

Two myosin heavy chains (MyHC) are expressed in the mammalian heart and are differentially regulated during development. In the mouse, the α-MyHC is expressed constitutively in the atrium. At birth, the β-MyHC is downregulated and replaced by the α-MyHC, which is the sole cardiac MyHC isoform in the adult heart. We have employed transgenic and gene-targeting methodologies to study the regulation of cardiac MyHC gene expression and the functional and developmental consequences of altered α-MyHC expression in the mouse.We previously characterized an α-MyHC promoter capable of driving tissue-specific and developmentally correct expression of a CAT (chloramphenicol acetyltransferase) marker in the mouse. Tissue surveys detected a small amount of CAT activity in the lung (Fig. 1a). The results of in situ hybridization analyses indicated that the pattern of CAT transcript in the adult heart (Fig. 1b, top panel) is the same as that of α-MyHC (Fig. 1b, lower panel). The α-MyHC gene is expressed in a layer of cardiac muscle (pulmonary myocardium) associated with the pulmonary veins (Fig. 1c). These studies extend our understanding of α-MyHC expression and delimit a third cardiac compartment.

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