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.

Autonomous differentiation of dorsal axial structures from an animal cap cleavage stage blastomere in Xenopus

Development ◽  
1991 ◽  
Vol 112 (4) ◽  
pp. 1103-1114 ◽  
Author(s):  
B.C. Gallagher ◽  
A.M. Hainski ◽  
S.A. Moody
Keyword(s):  
Original Position ◽  
Cleavage Stage ◽  
Cell Stage ◽  
Dorsal Midline ◽  
Developmental Program ◽  
Opposite Pole ◽  
Secondary Axis ◽  
Dorsal Axis ◽  
Exogenous Growth ◽  
Primary Axis

Dorsal or ventral blastomeres of the 16- and 32-cell stage animal hemisphere were labeled with a lineage dye and transplanted into the position of a ventral, vegetal midline blastomere. The donor blastomeres normally give rise to substantial amounts of head structures and central nervous system, whereas the blastomere which they replaced normally gives rise to trunk mesoderm and endoderm. The clones derived from the transplanted ventral blastomeres were found in tissues appropriate for their new position, whereas those derived from the transplanted dorsal blastomeres were found in tissues appropriate for their original position. The transplanted dorsal clones usually migrated into the host's primary axis (D1.1, 92%; D1.1.1, 69%; D1.1.2, 100%), and in many cases they also induced and populated a secondary axis (D1.1, 43%; D1.1.1, 67%; D1.1.2, 63%). Bilateral deletion of the dorsal blastomeres resulted in partial deficits of dorsal axial structures in the majority of cases, whereas deletions of ventral midline blastomeres did not. When the dorsal blastomeres were cultured as explants they elongated. Notochord and cement glands frequently differentiated in these explants. These studies show that the progeny of the dorsal, midline, animal blastomeres: (1) follow their normal lineage program to populate dorsal axial structures after the blastomere is transplanted to the opposite pole of the embryo; (2) induce and contribute to a secondary axis from their transplanted position in many embryos; (3) are important for the normal formation of the entire length of the dorsal axis; and (4) autonomously differentiate in the absence of exogenous growth factor signals. These data indicate that by the 16-cell stage, these blastomeres have received instructions regarding their fate, and they are intrinsically capable of carrying out some of their developmental program.

Cortical cytoplasm, which induces dorsal axis formation in Xenopus, is inactivated by UV irradiation of the oocyte

Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 277-285 ◽  
Author(s):  
T. Holowacz ◽  
R.P. Elinson
Keyword(s):  
Spatial Distribution ◽  
Uv Irradiation ◽  
Axis Formation ◽  
Prophase I ◽  
Xenopus Embryos ◽  
Secondary Axis ◽  
Vegetal Cortex ◽  
Dorsal Axis ◽  
Cell Embryo ◽  
Maternal Determinants

Localized maternal determinants control the formation of dorsal axial structures in Xenopus embryos. To examine the spatial distribution of dorsal determinants, we injected cytoplasm from various regions of the egg and 16-cell embryo into the ventral vegetal cells of a 16-cell recipient embryo. Cortical cytoplasm from the egg vegetal surface induced the formation of a secondary dorsal axis in 53% of recipients. In contrast, animal cortical, equatorial cortical and vegetal deep cytoplasm never induced secondary axis formation. We also compared the axis-inducing ability of animal versus vegetal dorsal cortical cytoplasm from 16-cell embryos. Significantly more dorsalizing activity was found in vegetal dorsal cytoplasm compared to animal dorsal cytoplasm at this stage. Previous work has shown that UV irradiation of the vegetal surface of either prophase I oocytes, or fertilized eggs, leads to the development of embryos that lack dorsal structures. Egg vegetal cortical cytoplasm was capable of restoring the dorsal axis of 16-cell recipient embryos derived from UV-irradiated oocytes or fertilized eggs. We also tested the axis inducing ability of cytoplasm obtained when UV-irradiated oocytes and eggs were treated as donors of cytoplasm. While vegetal cortical cytoplasm from UV-irradiated fertilized eggs retains its dorsalizing activity, cytoplasm obtained from eggs, UV irradiated as oocytes, does not. The egg vegetal cortex provides a suitable source for the isolation of maternal dorsal determinants. In addition, since UV irradiation of the oocyte vegetal surface destroys the dorsalizing activity of transferred cytoplasm, UV can be used to further restrict possible candidates for such determinants.

A cytoplasmic determinant for dorsal axis formation in an early embryo of Xenopus laevis

Development ◽  
1990 ◽  
Vol 110 (4) ◽  
pp. 1051-1056 ◽  
Author(s):  
M. Yuge ◽  
Y. Kobayakawa ◽  
M. Fujisue ◽  
K. Yamana
Keyword(s):  
Xenopus Laevis ◽  
Direct Evidence ◽  
Early Embryo ◽  
Dorsal Side ◽  
Axis Formation ◽  
Secondary Axis ◽  
Dorsal Axis ◽  
Cell Embryo ◽  
Dorsal Cells ◽  
Cytoplasmic Determinant

In Xenopus laevis, dorsal cells that arise at the future dorsal side of an early cleaving embryo have already acquired the ability to cause axis formation. Since the distribution of cytoplasmic components is markedly heterogeneous in an egg and embryo, it has been supposed that the dorsal cells are endowed with the activity to form axial structures by inheriting a unique cytoplasmic component or components localized in the dorsal region of an egg or embryo. However, there has been no direct evidence for this. To examine the activity of the cytoplasm of dorsal cells, we injected cytoplasm (dorsal cytoplasm) from dorsal vegetal cells of a Xenopus 16-cell embryo into ventral vegetal cells of a simultaneous recipient. The cytoplasm caused secondary axis formation in 42% of recipients. Histological examination revealed that well-developed secondary axes included notochord, as well as a neural tube and somites. However, injection of cytoplasm of ventral vegetal cells never caused secondary axis and most recipients became normal tailbud embryos. Furthermore, about two-thirds of ventral isolated halves injected with dorsal cytoplasm formed axial structures. These results show that dorsal, but not ventral, cytoplasm contains the component or components responsible for axis formation. This can be the first step towards identifying the molecular basis of dorsal axis formation.

Regulation of Spemann organizer formation by the intracellular kinase Xgsk-3

Development ◽  
1995 ◽  
Vol 121 (3) ◽  
pp. 755-765 ◽  
Author(s):  
S.B. Pierce ◽  
D. Kimelman
Keyword(s):  
Ectopic Expression ◽  
Dominant Negative ◽  
Dominant Negative Effect ◽  
Axis Formation ◽  
Serine Threonine Kinase ◽  
Spemann Organizer ◽  
Isolation And Characterization ◽  
Secondary Axis ◽  
Dorsal Axis ◽  
Negative Effect

Dorsal axis formation in the Xenopus embryo can be induced by the ectopic expression of several Wnt family members. In Drosophila, the protein encoded by the Wnt family gene, wingless, signals through a pathway that antagonizes the effects of the serine/threonine kinase zeste-white 3/shaggy. We describe the isolation and characterization of a Xenopus homolog of zeste-white 3/shaggy, Xgsk-3. A kinase-dead mutant of Xgsk-3, Xgsk-3K-->R, has a dominant negative effect and mimics the ability of Wnt to induce a secondary axis by induction of an ectopic Spemann organizer. Xgsk-3K-->R, like Wnt, induces dorsal axis formation when expressed in the deep vegetal cells, which do not contribute to the axis. These results indicate that the dorsal fate is actively repressed by Xgsk-3, which must be inactivated for dorsal axis formation to occur. Furthermore, our work suggests that the effects of Xgsk-3K-->R are mediated by an additional intercellular signal.

The vegetal determinants required for the Spemann organizer move equatorially during the first cell cycle

Development ◽  
1996 ◽  
Vol 122 (7) ◽  
pp. 2207-2214 ◽  
Author(s):  
M. Sakai
Keyword(s):  
Cell Cycle ◽  
Formation Process ◽  
Lithium Treatment ◽  
Axis Formation ◽  
Cell Stage ◽  
Spemann Organizer ◽  
Dorsal Axis ◽  
Vegetal Pole ◽  
Organizing Center ◽  
Cell Embryo

Embryos with no dorsal axis were obtained when more than 15% of the egg surface was deleted from the vegetal pole of the early 1-cell embryo of Xenopus laevis. The timing of the deletion in the first cell cycle was critical: dorsal-deficient embryos were obtained when the deletion began before time 0.5 (50% of the first cell cycle) whereas normal dorsal axis usually formed when the deletion was done later than time 0.8. The axis deficiency could be restored by lithium treatment and the injection of vegetal but not animal cytoplasm. Bisection of the embryo at the 2-cell stage, which is known to restore the dorsal structures in the UV-ventralized embryos, had no effect on the vegetal-deleted embryos. These results show clearly that, in Xenopus, (1) the dorsal determinants (DDs) localized in the vegetal pole region at the onset of development are necessary for dorsal axis development and (2) the DDs move from the vegetal pole to a subequatorial region where they are incorporated into gastrulating cells to form the future organizing center. A model for the early axis formation process in Xenopus is proposed.

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.

Maternal beta-catenin establishes a ‘dorsal signal’ in early Xenopus embryos

Development ◽  
1996 ◽  
Vol 122 (10) ◽  
pp. 2987-2996 ◽  
Author(s):  
C. Wylie ◽  
M. Kofron ◽  
C. Payne ◽  
R. Anderson ◽  
M. Hosobuchi ◽  
...  
Keyword(s):  
Glycogen Synthase ◽  
Axis Formation ◽  
Beta Catenin ◽  
Cell Stage ◽  
Midblastula Transition ◽  
Xenopus Embryos ◽  
Ability To Act ◽  
Zygotic Transcription ◽  
Spatial Terms ◽  
Marginal Zones

In previous work, we demonstrated that maternally encoded beta-catenin, the vertebrate homolog of armadillo, is required for formation of dorsal axial structures in early Xenopus embryos (Heasman, J., Crawford, A., Goldstone, K., Garner-Hamrick, P., Gumbiner, B., Kintner, C., Yoshida-Noro, C. and Wylie, C. (1994). Cell 79, 791–803). Here we investigated, firstly, the role(s) of beta-catenin in spatial terms, in different regions of the embryo, by injecting beta-catenin mRNA into individual blastomeres of beta-catenin-depleted embryos at the 32 cell stage. The results indicate that beta-catenin can rescue the dorsal axial structures in a non-cell-autonomous way and without changing the fates of the injected cells. This suggests that cells overexpressing beta-catenin send a ‘dorsal signal’ to other cells. This was confirmed by showing that beta-catenin overexpressing animal caps did not cause wild-type caps to form mesoderm, but did cause isolated beta-catenin-deficient marginal zones to form dorsal mesoderm. Furthermore beta-catenin-deficient vegetal masses treated with overexpressing caps regained their ability to act as Nieuwkoop Centers. Secondly, we studied the temporal activity of beta-catenin. We showed that zygotic transcription of beta-catenin starts after the midblastula transition (MBT), but does not rescue dorsal axial structures. We further demonstrated that the vegetal mass does not release a dorsal signal until after the onset of transcription, at the midblastula stage, suggesting that maternal beta-catenin protein is required at or before this time. Thirdly we investigated where, in relationship to other gene products known to be active in axis formation, beta-catenin is placed. We find that BVg1, bFGF, tBR (the truncated form of BMP2/4R), siamois and noggin activities are all downstream of beta-catenin, as shown by the fact that injection of their mRNAs rescues the effect of depleting maternally encoded beta-catenin. Interference with the action of glycogen synthase kinase (GSK), a vertebrate homolog of the Drosophila gene product, zeste white 3 kinase, does not rescue the effect, suggesting that it is upstream.

XIPOU 2, a noggin-inducible gene, has direct neuralizing activity

Development ◽  
1995 ◽  
Vol 121 (3) ◽  
pp. 721-730 ◽  
Author(s):  
S.E. Witta ◽  
V.R. Agarwal ◽  
S.M. Sato
Keyword(s):  
Cell Fate ◽  
Neural Induction ◽  
Mesoderm Induction ◽  
Class Iii ◽  
Cell Stage ◽  
Neuronal Phenotype ◽  
Pou Domain ◽  
Spemann's Organizer ◽  
Stage Embryo ◽  
Synthetic Mrna

XIPOU 2, a member of the class III POU domain family, is expressed initially in Spemann's organizer, and later, in discrete regions of the developing nervous system in Xenopus laevis. XIPOU 2 may act downstream from initial neural induction events, since it is activated by the neural inducer, noggin. To determine if XIPOU 2 participates in the early events of neurogenesis, synthetic mRNA was microinjected into specific blastomeres of the 32-cell stage embryo. Misexpression of XIPOU 2 in the epidermis causes a direct switch in cell fate from an epidermal to a neuronal phenotype. In the absence of mesoderm induction, XIPOU 2 has the ability to induce a neuronal phenotype in uncommitted ectoderm. These data demonstrate the potential of XIPOU 2 to act as a master regulator of neurogenesis.

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