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.

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.

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.

scholarly journals Reducing inositol lipid hydrolysis, Ins(1,4,5)P3 receptor availability, or Ca2+ gradients lengthens the duration of the cell cycle in Xenopus laevis blastomeres.

1992 ◽  
Vol 116 (1) ◽  
pp. 147-156 ◽  
Author(s):  
J K Han ◽  
K Fukami ◽  
R Nuccitelli
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Xenopus Laevis ◽  
Mitotic Cell ◽  
Embryonic Cell ◽  
Cycle Duration ◽  
Cell Stage ◽  
Stage Embryo ◽  
Rate Of Hydrolysis ◽  
Xenopus Laevis Embryos

We have microinjected a mAb specifically directed to phosphatidylinositol 4,5-bisphosphate (PIP2) into one blastomere of two-cell stage Xenopus laevis embryos. This antibody binds to endogenous PIP2 and reduces its rate of hydrolysis by phospholipase C. Antibody-injected blastomeres undergo partial or complete arrest of the cell cycle whereas the uninjected sister blastomeres divided normally. Since PIP2 hydrolysis normally produces diacylglycerol (DG) and inositol 1,4,5-triphosphate (Ins[1,4,5]P3), we attempted to measure changes in the levels of DG following stimulation of PIP2 hydrolysis in antibody-injected oocytes. The total amount of DG in antibody-injected oocytes was significantly reduced compared to that of water-injected ones following stimulation by either acetylcholine or progesterone indicating that the antibody does indeed suppress PIP2 hydrolysis. We also found that the PIP2 antibodies greatly reduced the amount of intracellular Ca2+ released in the egg cortex during egg activation. As an indirect test for Ins(1,4,5)P3 involvement in the cell cycle we injected heparin which competes with Ins(1,4,5)P3 for binding to its receptor, and thus inhibits Ins(1,4,5)P3-induced Ca2+ release. Microinjection of heparin into one blastomere of the two-cell stage embryo caused partial or complete arrest of the cell cycle depending upon the concentration of heparin injected. We further investigated the effect of reducing any [Ca2+]i gradients by microinjecting dibromo-BAPTA into the blastomere. Dibromo-BAPTA injection completely blocked mitotic cell division when a final concentration of 1.5 mM was used. These results suggest that PIP2 turnover as well as second messenger activity influence cell cycle duration during embryonic cell division in frogs.

scholarly journals Fate map for the 32-cell stage of Xenopus laevis

Development ◽  
1987 ◽  
Vol 99 (4) ◽  
pp. 527-551 ◽  
Author(s):  
L. Dale ◽  
J.M. Slack
Keyword(s):  
Xenopus Laevis ◽  
Short Range ◽  
Three Dimensional ◽  
Spatial Localization ◽  
Spatial Location ◽  
Fate Map ◽  
Cell Stage ◽  
Range Cell ◽  
Early Gastrula ◽  
Xenopus Laevis Embryos

A complete fate map has been produced for the 32-cell stage of Xenopus laevis. Embryos with a regular cleavage pattern were selected and individual blastomeres were injected with the lineage label fluorescein-dextran-amine (FDA). The spatial location of the clones was deduced from three-dimensional (3D) reconstructions of later stages and the volume of each tissue colonized by labelled cells in each tissue was measured. The results from 107 cases were pooled to give a fate map which shows the fate of each blastomere in terms of tissue types, the composition of each tissue by blastomere, the location of each prospective region on the embryo and the fate of each blastomere in terms of spatial localization. Morphogenetic movements up to stage 10 (early gastrula) were assessed by carrying out a number of orthotopic grafts at blastula and gastrula stages using donor embryos uniformly labelled with FDA. Although there is a regular topographic projection from the 32-cell stage this varies a little between individuals because of variability of positions of cleavage planes and because of short-range cell mixing during gastrulation. The cell mixing means that the topographic projection fails for anteroposterior segments of the dorsal axial structures and it is not possible to include short segments of notochord or neural tube or individual somites on the pregastrulation fate map.

Xenopus maternal RNAs from a dorsal animal blastomere induce a secondary axis in host embryos

Development ◽  
1992 ◽  
Vol 116 (2) ◽  
pp. 347-355 ◽  
Author(s):  
A.M. Hainski ◽  
S.A. Moody
Keyword(s):  
Gene Families ◽  
Similar Process ◽  
Axis Formation ◽  
Cell Stage ◽  
Dorsal Midline ◽  
Rna Molecules ◽  
Secondary Axis ◽  
Dorsal Axis ◽  
Stage Embryo ◽  
Maternal Determinants

The initial steps of dorsal axis formation are controlled by localized maternal determinants in Drosophila, and a similar process has been proposed in Xenopus. The present study demonstrates that there are axis-inducing RNA molecules located in a specific dorsal midline, animal blastomere (D1.1) of the 16-cell-stage embryo. This blastomere, although in the animal hemisphere at cleavage stages, populates most of the dorsal lip of the blastopore, the region of Spemann's organizer, during gastrulation, and is the major progenitor for dorsal mesodermal tissues. Cytosol from this blastomere causes ventral cells to take a more dorsal fate. RNA from this blastomere induces a secondary axis when injected into ventral blastomeres and restores the dorsal axis in UV-irradiated embryos. In Xenopus, activin beta B, goosecoid and Xwnt-8 RNAs can ectopically induce a dorsal axis; however, none is a maternal transcript. Therefore, the D1.1 blastomere probably contains dorsal determinant(s) that are either maternal members of these gene families, or other presently unknown molecule(s). Regardless of the identity of the determinant(s), this study presents the first indication that Xenopus maternal RNAs in the dorsal animal hemisphere are able to organize the dorsal axis.

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.

Initiation of mesodermal cell migration and spreading relative to gastrulation in the urodele amphibian Pleurodeles waltl

Development ◽  
1989 ◽  
Vol 105 (2) ◽  
pp. 351-363 ◽  
Author(s):  
D.-L. Shi ◽  
T. Darribere ◽  
K.E. Johnson ◽  
J.-C. Boucaut
Keyword(s):  
Cell Migration ◽  
Marginal Zone ◽  
Gastrula Stage ◽  
Cell Stage ◽  
Animal Pole ◽  
Mesodermal Cell ◽  
Marginal Cells ◽  
Pleurodeles Waltl ◽  
Early Gastrula

We have investigated the autonomous migration of marginal cells and their interactions with extracellular matrix (ECM) located on the inner surface of the blastocoel roof in the urodele amphibian, Pleurodeles waltl, using a novel in vitro migration assay. Animal hemispheres containing equatorial cells removed at different cleavage stages and dorsal marginal zone (DMZ) explants of early gastrula stage were cultured either on fibronectin (FN)-coated or ECM-conditioned substrata. In explanted animal hemispheres, dorsal marginal cells showed autonomous migration on FN-coated substratum at the same time as the onset of gastrulation in control embryos. They acquired this capacity at least at the 32-cell stage, whereas lateral and ventral marginal cells acquired it after the 64-cell stage. DMZ outgrowths of early gastrula stage exhibited autonomous spreading on both substrata. In addition, we showed that they spread preferentially toward the animal pole when deposited on substratum conditioned by the dorsal roof of the blastocoel. By culturing dissociated marginal cells on ECM- conditioned substratum, we also found that increased spreading capacity of marginal cells was related to the initiation of their migration. A comparative study of the migration of marginal cells in ultraviolet (u.v.)-irradiated and normal embryos was also made. The results indicate that dorsal marginal cell migration was absent or dramatically reduced by u.v.-irradiation. These results suggest that the differential acquisition in the spreading capacity both in timing and in intensity around the marginal zone was correlated with the sequential involution of mesodermal cells in the course of gastrulation.

The origins of primitive blood in Xenopus: implications for axial patterning

Development ◽  
1999 ◽  
Vol 126 (3) ◽  
pp. 423-434 ◽  
Author(s):  
M.C. Lane ◽  
W.C. Smith
Keyword(s):  
Xenopus Laevis ◽  
Marginal Zone ◽  
Cell Stage ◽  
Axial Patterning ◽  
Xenopus Embryos ◽  
Spemann's Organizer ◽  
Dorsal Mesoderm

The marginal zone in Xenopus laevis is proposed to be patterned with dorsal mesoderm situated near the upper blastoporal lip and ventral mesoderm near the lower blastoporal lip. We determined the origins of the ventralmost mesoderm, primitive blood, and show it arises from all vegetal blastomeres at the 32-cell stage, including blastomere C1, a progenitor of Spemann's organizer. This demonstrates that cells located at the upper blastoporal lip become ventral mesoderm, not solely dorsal mesoderm as previously believed. Reassessment of extant fate maps shows dorsal mesoderm and dorsal endoderm descend from the animal region of the marginal zone, whereas ventral mesoderm descends from the vegetal region of the marginal zone, and ventral endoderm descends from cells located vegetal of the bottle cells. Thus, the orientation of the dorsal-ventral axis of the mesoderm and endoderm is rotated 90(degrees) from its current portrayal in fate maps. This reassessment leads us to propose revisions in the nomenclature of the marginal zone and the orientation of the axes in pre-gastrula Xenopus embryos.

scholarly journals Imaging patterns of calcium transients during neural induction in Xenopus laevis embryos

2000 ◽  
Vol 113 (19) ◽  
pp. 3519-3529 ◽  
Author(s):  
C. Leclerc ◽  
S.E. Webb ◽  
C. Daguzan ◽  
M. Moreau ◽  
A.L. Miller
Keyword(s):  
Xenopus Laevis ◽  
Calcium Signalling ◽  
Neural Induction ◽  
Calcium Transients ◽  
Free Calcium ◽  
Treated Animal ◽  
Cell Stage ◽  
Subsequent Formation ◽  
Xenopus Laevis Embryos

Through the injection of f-aequorin (a calcium-sensitive bioluminescent reporter) into the dorsal micromeres of 8-cell stage Xenopus laevis embryos, and the use of a Photon Imaging Microscope, distinct patterns of calcium signalling were visualised during the gastrulation period. We present results to show that localised domains of elevated calcium were observed exclusively in the anterior dorsal part of the ectoderm, and that these transients increased in number and amplitude between stages 9 to 11, just prior to the onset of neural induction. During this time, however, no increase in cytosolic free calcium was observed in the ventral ectoderm, mesoderm or endoderm. The origin and role of these dorsal calcium-signalling patterns were also investigated. Calcium transients require the presence of functional L-type voltage-sensitive calcium channels. Inhibition of channel activation from stages 8 to 14 with the specific antagonist R(+)BayK 8644 led to a complete inhibition of the calcium transients during gastrulation and resulted in severe defects in the subsequent formation of the anterior nervous system. BayK treatment also led to a reduction in the expression of Zic3 and geminin in whole embryos, and of NCAM in noggin-treated animal caps. The possible role of calcium transients in regulating developmental gene expression is discussed.

Structure and developmental tendency of the dorsal marginal zone in the early amphibian gastrula

Development ◽  
1971 ◽  
Vol 25 (2) ◽  
pp. 263-276
Author(s):  
Nobushige Ikushima ◽  
Setsuko Maruyama
Keyword(s):  
Outer Layer ◽  
Marginal Zone ◽  
Kinetic Properties ◽  
Electron Microscopic ◽  
Structural Differences ◽  
Peripheral Surface ◽  
Early Gastrula ◽  
A Share

The peripheral surface of a fertilized, uncleaved egg is subdivided through cleavage and is allotted to constituent cells. This is called the primary surface. In an early morula a constituent cell has two kinds of surfaces: the primary surface, and the secondary surface, which does not participate in forming the periphery of the embryo. Electron-microscopic observations showed structural differences between the two surfaces. When the dorsal marginal zone of an early gastrula of Hynobius nebulosus is excised and immersed in Feldman's solution, the piece can easily be separated into two layers: the outer layer, whose constituent cells are given a share of the primary surface, and the inner layer, whose constituent cells are completely covered only by the secondary surface. Both an explanted piece of the outer layer and an intact double-layered piece show three kinds of movement: spreading, convergence followed by stretching, and spherical thickening. The inner layer is kinetically very inert, showing slight spreading and thickening. An explanted piece of the outer layer differentiates into axial mesodermal structures, while the inner layer does not. When a piece of either the inner or the outer layer is implanted in the blastocoel of another gastrula, it induces deuterencephalic and spino-caudal structures and seems to differentiate into axial mesodermal structures. Differences of kinetic properties and differentiation are considered to result from the fact that the outer layer has the primary surface, while the inner layer does not. Functional effects of the primary surface on the movement of tissues and differentiation are discussed.

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