Vegetal rotation, a new gastrulation movement involved in the internalization of the mesoderm and endoderm in Xenopus

Development ◽  
1999 ◽  
Vol 126 (16) ◽  
pp. 3703-3713 ◽  
Author(s):  
R. Winklbauer ◽  
M. Schurfeld
Keyword(s):  
Driving Force ◽  
Marginal Zone ◽  
Cell Mass ◽  
Fundamental Importance ◽  
Major Step ◽  
Animal Pole ◽  
Main Achievement ◽  
Internalization Process ◽  
Gastrulation Movement ◽  
Vegetal Cell

A main achievement of gastrulation is the movement of the endoderm and mesoderm from the surface of the embryo to the interior. Despite its fundamental importance, this internalization process is not well understood in amphibians. We show that in Xenopus, an active distortion of the vegetal cell mass, vegetal rotation, leads to a dramatic expansion of the blastocoel floor and a concomitant turning around of the marginal zone which constitutes the first and major step of mesoderm involution. This vigorous inward surging of the vegetal region into the blastocoel can be analyzed in explanted slices of the gastrula, and is apparently driven by cell rearrangement. Thus, the prospective endoderm, previously thought to be moved passively, provides the main driving force for the internalization of the mesendoderm during the first half of gastrulation. For further involution, and for normal positioning of the involuted mesoderm and its rapid advance toward the animal pole, fibronectin-independent interaction with the blastocoel roof is required.

The role of fibroblast growth factor in early Xenopus development

Development ◽  
1989 ◽  
Vol 107 (Supplement) ◽  
pp. 141-148 ◽  
Author(s):  
J. M. W. Slack ◽  
B. G. Darlington ◽  
L. L. Gillespie ◽  
S. F. Godsave ◽  
H. V. Isaacs ◽  
...  
Keyword(s):  
Marginal Zone ◽  
Specific Activity ◽  
Progressive Increase ◽  
Defined Medium ◽  
Relative Molecular Mass ◽  
Heparin Binding ◽  
Cell Stage ◽  
Animal Pole

In early amphibian development, the mesoderm is formed around the equator of the blastula in response to an inductive signal from the endoderm. A screen of candidate substances showed that a small group of heparin-binding growth factors (HBGFs) were active as mesoderm-inducing agents in vitro. The factors aFGF, bFGF, kFGF and ECDGF all show similar potency and can produce inductions at concentrations above about 100 pM. The product of the murine int-2 gene is also active, but with a lower specific activity. Above the induction threshold there is a progressive increase of muscle formation with dose. Single blastula ectoderm cells can be induced and will differentiate in a defined medium to form mesodermal tissues. All inner blastula cells are competent to respond to the factors but outer cells, bearing oocyte-derived membrane, are not. Inducing activity can be extracted from Xenopus blastulae and binds to heparin like the previously described HBGFs. Antibody neutralization and Western blotting experiments identify this activity as bFGF. The amounts present are small but would be sufficient to evoke inductions in vivo. It is not yet known whether the bFGF is localized to the endoderm, although it is known that inducing activity secreted by endodermal cells can be neutralized by heparin. The competence of ectoderm to respond to HBGFs rises from about the 128-cell stage and falls again by the onset of gastrulation. This change is paralleled by a rise and fall of binding of 125I-aFGF. Chemical cross-linking reveals that this binding is attributable to a receptor of relative molecular mass about 130 × 103. The receptor is present both in the marginal zone, which responds to the signal in vivo, and in the animal pole region, which is not induced in vivo but which will respond to HBGFs in vitro. In the embryo, the induction in the vicinity of the dorsal meridian is much more potent than that around the remainder of the marginal zone circumference. Dorsal inductions contain notochord and will dorsalize ventral mesoderm with which they are later placed in contact. This effect might be due to a local high bFGF concentration or, more likely, to the secretion in the dorsal region of an additional, synergistic factor. It is known that TGF-β-1 and -2 can greatly increase the effect of low doses of bFGF, although it has not yet been demonstrated that they are present in the embryo. Lithium salts have a dorsalizing effect on whole embryos or on explants from the ventral marginal zone, and also show potent synergism when applied together with HBGFs.

Xmsx-1 modifies mesodermal tissue pattern along dorsoventral axis in Xenopus laevis embryo

Development ◽  
1997 ◽  
Vol 124 (13) ◽  
pp. 2553-2560 ◽  
Author(s):  
R. Maeda ◽  
A. Kobayashi ◽  
R. Sekine ◽  
J.J. Lin ◽  
H. Kung ◽  
...  
Keyword(s):  
Target Gene ◽  
Marginal Zone ◽  
Dominant Negative ◽  
Gastrula Stage ◽  
Cell Stage ◽  
Animal Pole ◽  
Alpha Globin ◽  
Molecular Marker Analysis ◽  
Alpha Actin ◽  
Stage Stage

This study analyzes the expression and the function of Xenopus msx-1 (Xmsx-1) in embryos, in relation to the ventralizing activity of bone morphogenetic protein-4 (BMP-4). Expression of Xmsx-1 was increased in UV-treated ventralized embryos and decreased in LiCl-treated dorsalized embryos at the neurula stage (stage 14). Whole-mount in situ hybridization analysis showed that Xmsx-1 is expressed in marginal zone and animal pole areas, laterally and ventrally, but not dorsally, at mid-gastrula (stage 11) and late-gastrula (stage 13) stages. Injection of BMP-4 RNA, but not activin RNA, induced Xmsx-1 expression in the dorsal marginal zone at the early gastrula stage (stage 10+), and introduction of a dominant negative form of BMP-4 receptor RNA suppressed Xmsx-1 expression in animal cap and ventral marginal zone explants at stage 14. Thus, Xmsx-1 is a target gene specifically regulated by BMP-4 signaling. Embryos injected with Xmsx-1 RNA in dorsal blastomeres at the 4-cell stage exhibited a ventralized phenotype, with microcephaly and swollen abdomen. Histological observation and immunostaining revealed that these embryos had a large block of muscle tissue in the dorsal mesodermal area instead of notochord. On the basis of molecular marker analysis, however, the injection of Xmsx-1 RNA did not induce the expression of alpha-globin, nor reduce cardiac alpha-actin in dorsal marginal zone explants. Furthermore, a significant amount of alpha-actin was induced and alpha-globin was turned off in the ventral marginal zone explants injected with Xmsx-1. These results indicated that Xmsx-1 is a target gene of BMP-4 signaling, but possesses a distinct activity on dorsal-ventral patterning of mesodermal tissues.

Mesoderm induction and the control of gastrulation in Xenopus laevis: the roles of fibronectin and integrins

Development ◽  
1990 ◽  
Vol 108 (2) ◽  
pp. 229-238 ◽  
Author(s):  
J.C. Smith ◽  
K. Symes ◽  
R.O. Hynes ◽  
D. DeSimone
Keyword(s):  
Marginal Zone ◽  
Single Cells ◽  
Mesoderm Induction ◽  
Convergent Extension ◽  
Animal Pole ◽  
Post Translational Modifications ◽  
Coated Surface ◽  
Pole Cells ◽  
And Control ◽  
Future Work

Exposure of isolated Xenopus animal pole ectoderm to the XTC mesoderm-inducing factor (XTC-MIF) causes the tissue to undergo gastrulation-like movements. In this paper, we take advantage of this observation to investigate the control of various aspects of gastrulation in Xenopus. Blastomeres derived from induced animal pole regions are able, like marginal zone cells, but unlike control animal pole blastomeres, to spread and migrate on a fibronectin-coated surface. Dispersed animal pole cells are also able to respond to XTC-MIF in this way; this is one of the few mesoderm-specific responses to induction that has been observed in single cells. The ability of induced animal pole cells to spread on fibronectin is abolished by the peptide GRGDSP. However, the elongation of intact explants is unaffected by this peptide. This may indicate that fibronectin-mediated cell migration is not required for convergent extension. We have investigated the molecular basis of XTC-MIF-induced gastrulation-like movements by measuring rates of synthesis of fibronectin and of the integrin beta 1 chain in induced and control explants. No significant differences were observed, and this suggests that gastrulation is not initiated simply by control of synthesis of these molecules. In future work, we intend to investigate synthesis of other integrin subunits and to examine possible post-translational modifications to fibronectin and the integrins.

Experimental analysis of the extension of the dorsal marginal zone in Pleurodeles waltl gastrulae

Development ◽  
1987 ◽  
Vol 100 (1) ◽  
pp. 147-161 ◽  
Author(s):  
D.L. Shi ◽  
M. Delarue ◽  
T. Darribere ◽  
J.F. Riou ◽  
J.C. Boucaut
Keyword(s):  
Electron Microscopy ◽  
Marginal Zone ◽  
Cell Lineage ◽  
Single Layer ◽  
Gastrula Stage ◽  
Animal Pole ◽  
A Cell ◽  
Pleurodeles Waltl ◽  
Scanning Electron

The capacity for extension of the dorsal marginal zone (DMZ) in Pleurodeles waltl gastrulae was studied by scanning electron microscopy and grafting experiments. At the onset of gastrulation, the cells of the animal pole (AP) undergo important changes in shape and form a single layer. As gastrulation proceeds, the arrangement of cells also changes in the noninvoluted DMZ: radial intercalation leads to a single layer of cells. Grafting experiments involving either AP or DMZ explants were performed using a cell lineage tracer. When rotated 90 degrees or 180 degrees, grafted DMZ explants were able to involute normally and there was extension according to the animal-vegetal axis of the host. In contrast, neither single nor bilayered explants from AP involutes completely, and neither extends when grafted in place of the DMZ. Furthermore, when inside of the host, these AP grafts curl up and inhibit the closure of the blastopore. Once transplanted to the AP region, the DMZ showed no obvious autonomous extension. DMZs cultured in vitro showed little extension and this only from the late gastrula stage onward. Removal of blastocoel roof blocked involution to a varied extent, depending on the developmental stage of the embryos. From these results, it is argued that differences could well exist in the mechanism of gastrulation between anuran and urodele embryos. That migrating mesodermal cells play a major role in urodele gastrulation is discussed.

Analysis of competence: receptors for fibroblast growth factor in early Xenopus embryos

Development ◽  
1989 ◽  
Vol 106 (1) ◽  
pp. 203-208 ◽  
Author(s):  
L.L. Gillespie ◽  
G.D. Paterno ◽  
J.M. Slack
Keyword(s):  
Binding Sites ◽  
Marginal Zone ◽  
Developmental Competence ◽  
Relative Molecular Mass ◽  
Scatchard Analysis ◽  
Animal Pole ◽  
Basic Fgf ◽  
Affinity Labelling ◽  
Pole Cells ◽  
Mesodermal Tissue

Xenopus ectodermal cells have previously been shown to respond to acidic and basic FGF by differentiating into mesodermal tissue. In the present study, ectodermal explants from Xenopus blastulae were shown to have high affinity binding sites for 125I-aFGF (Kd = 1.4 X 10(−10) M). The total number of sites, determined by Scatchard analysis, was 3 X 10(8) per explant (surface area of approximately 1 mm2). Two putative receptors of relative molecular mass 130,000 and 140,000 were identified by chemical crosslinking to 125I-aFGF. Both acidic and basic FGF, but not TGF beta 2, could compete for affinity labelling of these bands. The receptor density at the cell surface parallels the developmental competence of Xenopus animal pole cells to respond to FGF. Receptors are present at highest density in the marginal zone but are not restricted to cells in this region.

SCL specifies hematopoietic mesoderm in Xenopus embryos

Development ◽  
1998 ◽  
Vol 125 (14) ◽  
pp. 2611-2620 ◽  
Author(s):  
P.E. Mead ◽  
C.M. Kelley ◽  
P.S. Hahn ◽  
O. Piedad ◽  
L.I. Zon
Keyword(s):  
Cell Fate ◽  
Marginal Zone ◽  
Dominant Negative ◽  
Treated Animal ◽  
Animal Pole ◽  
Over Expression ◽  
Targeted Gene Disruption ◽  
Helix Loop Helix ◽  
Absolute Requirement ◽  
Necessary And Sufficient

Targeted gene disruption experiments in the mouse have demonstrated an absolute requirement for several transcription factors for the development of hematopoietic progenitors during embryogenesis. Disruption of the basic helix-loop-helix gene SCL (stem cell leukemia) causes a block early in the hematopoietic program with defects in all hematopoietic lineages. To understand how SCL participates in the organogenesis of blood, we have isolated cDNAs encoding Xenopus SCL and characterized the function of SCL during embryogenesis. We demonstrate that SCL is expressed in ventral mesoderm early in embryogenesis. SCL expression is induced by BMP-4, and a dominant negative BMP-4 receptor inhibits SCL expression in the ventral region of the embryo. Expression of SCL in either bFGF-treated animal pole explants or dorsal marginal zone explants leads to the expression of globin protein. Furthermore, over-expression of SCL does not alter normal dorsal-ventral patterning in the embryo, indicating that SCL acts to specify mesoderm to a hematopoietic fate after inductive and patterning events have occurred. We propose that SCL is both necessary and sufficient to specify hematopoietic mesoderm, and that it has a similar role in specifying hematopoietic cell fate as MyoD has in specifying muscle cell fate.

Nuclear beta-catenin is required to specify vegetal cell fates in the sea urchin embryo

Development ◽  
1999 ◽  
Vol 126 (2) ◽  
pp. 345-357 ◽  
Author(s):  
C.Y. Logan ◽  
J.R. Miller ◽  
M.J. Ferkowicz ◽  
D.R. McClay
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Beta Catenin ◽  
Cell Fate Specification ◽  
Cell Fates ◽  
Cell Stage ◽  
Animal Pole ◽  
Fate Specification ◽  
The Relationship ◽  
Vegetal Cell

Beta-catenin is thought to mediate cell fate specification events by localizing to the nucleus where it modulates gene expression. To ask whether beta-catenin is involved in cell fate specification during sea urchin embryogenesis, we analyzed the distribution of nuclear beta-catenin in both normal and experimentally manipulated embryos. In unperturbed embryos, beta-catenin accumulates in nuclei that include the precursors of the endoderm and mesoderm, suggesting that it plays a role in vegetal specification. Using pharmacological, embryological and molecular approaches, we determined the function of beta-catenin in vegetal development by examining the relationship between the pattern of nuclear beta-catenin and the formation of endodermal and mesodermal tissues. Treatment of embryos with LiCl, a known vegetalizing agent, caused both an enhancement in the levels of nuclear beta-catenin and an expansion in the pattern of nuclear beta-catenin that coincided with an increase in endoderm and mesoderm. Conversely, overexpression of a sea urchin cadherin blocked the accumulation of nuclear beta-catenin and consequently inhibited the formation of endodermal and mesodermal tissues including micromere-derived skeletogenic mesenchyme. In addition, nuclear beta-catenin-deficient micromeres failed to induce a secondary axis when transplanted to the animal pole of uninjected host embryos, indicating that nuclear beta-catenin also plays a role in the production of micromere-derived signals. To examine further the relationship between nuclear beta-catenin in vegetal nuclei and micromere signaling, we performed both transplantations and deletions of micromeres at the 16-cell stage and demonstrated that the accumulation of beta-catenin in vegetal nuclei does not require micromere-derived cues. Moreover, we demonstrate that cell autonomous signals appear to regulate the pattern of nuclear beta-catenin since dissociated blastomeres possessed nuclear beta-catenin in approximately the same proportion as that seen in intact embryos. Together, these data show that the accumulation of beta-catenin in nuclei of vegetal cells is regulated cell autonomously and that this localization is required for the establishment of all vegetal cell fates and the production of micromere-derived signals.

Spatial and temporal properties of ventral blood island induction in Xenopus laevis

Development ◽  
1999 ◽  
Vol 126 (23) ◽  
pp. 5327-5337 ◽  
Author(s):  
G. Kumano ◽  
L. Belluzzi ◽  
W.C. Smith
Keyword(s):  
Marginal Zone ◽  
Selective Inhibition ◽  
Distinct Pattern ◽  
Dominant Negative ◽  
Animal Pole ◽  
Blood Island ◽  
Spemann Organizer ◽  
Temporal Properties ◽  
Vegetal Pole ◽  
Marginal Zones

Questions of dorsoventral axis determination and patterning in Xenopus seek to uncover the mechanisms by which particular mesodermal fates, for example somite, are specified in the dorsal pole of the axis while other mesoderm fates, for example, ventral blood island (VBI), are specified at the ventral pole. We report here that the genes Xvent-1, Xvent-2, and Xwnt-8 do not appear to be in the pathway of VBI induction, contrary to previous reports. Results from the selective inhibition of bone morphogenetic protein (BMP) activity, a key regulator of VBI induction, by ectopic Noggin, Chordin, or dominant negative BMP ligands and receptors suggest an alternative route of VBI induction. Injection of noggin or chordin RNA into animal pole blastomeres effectively inhibited VBI development, while marginal zone injection had no effect. Cell autonomous inhibition of BMP activity in epidermis with dominant negative ligand dramatically reduced the amount of (α)T3 globin expression. These results indicate that signaling activity from the Spemann Organizer alone may not be sufficient for dorsoventral patterning in the marginal zone and that an inductive interaction between presumptive VBIs and ectoderm late in gastrulation may be crucial. In agreement with these observations, other results show that in explanted blastula-stage marginal zones a distinct pattern develops with a restricted VBI-forming region at the vegetal pole that is independent of the patterning activity of the Spemann Organizer.

Interconnection Trends and Impact on Materials

1986 ◽  
Vol 72 ◽  
Author(s):  
W. H. Knausenberger ◽  
M. R. Pinnel
Keyword(s):  
Driving Force ◽  
Large Scale ◽  
Heat Dissipation ◽  
New Technology ◽  
Electronic Systems ◽  
Major Step ◽  
Technological Advances ◽  
Rapid Transition ◽  
To Come ◽  
And Performance

AbstractRapid technological advances in electronic systems technologies are placing increasingly severe demands on interconnection media. One primary driving force is the evolutionary advance in the scale of integration in silicon with its inherent cost and performance advantages. A second key driving force is the revolutionary development of photonics which is rapidly integrating into most levels of the interconnection heirarchy. The performance of large scale electronic systems will be increasingly dominated and limited by their interconnection environment. To sustain the present rate of growth in the performance of future systems, new technology directions in interconnection will be necessary.This paper will explore the traditional levels of interconnection from IC chip packages to the frame level. It will be shown how the various levels of interconnection interrelate and how all levels must be improved simultaneously to achieve the full performance and cost benefits offered by device advances and photonics. The first major step in this evolution is well underway with the rapid transition to surface mounting of devices. This places new demands on materials and assembly technologies which will be discussed. However, the demands of this first step may eventually appear to be minor compared to those yet to come if current trends continue. Several scenarios for this future will be considered and related to the challenges placed on interconnection technology hardware and materials in terms of performance characteristics such as density, speed and heat dissipation.

scholarly journals 113 THE DISTRIBUTION OF THE LEPTIN PROTEIN WITHIN BOVINE OOCYTES AND PRE-IMPLANTATION EMBRYOS MATURED AND FERTILIZED IN VITRO

2005 ◽  
Vol 17 (2) ◽  
pp. 207
Author(s):  
Z. Madeja ◽  
D. Lechniak ◽  
J. Peippo ◽  
M. Switonski
Keyword(s):  
Protein Localization ◽  
Cell Mass ◽  
Immunofluorescent Staining ◽  
Preimplantation Embryos ◽  
Trophoblast Invasion ◽  
Protein Distribution ◽  
Santa Cruz ◽  
Animal Pole ◽  
Bovine Oocytes

It has recently been documented that leptin regulates processes linked to reproduction including preimplantation development, embryo implantation (trophoblast invasion), and fetal growth. Transcripts for the leptin gene (LEP) and the leptin receptor gene (LEPR) have been identified in ovary, testis, placenta, endometrium, ovarian follicles, and oocytes, and also in mouse, rat, human, and bovine pre-implantation embryos. Moreover, the leptin protein was detected in mouse and human oocytes and embryos, and its localization was polarized. The distribution of regulatory proteins within oocytes and pre-implantation embryos is critical for early mammalian development, such as determination of the animal pole and the establishment of the trophoblast and the inner cell mass cells (ICM). So far there is no published evidence concerning this phenomenon in bovine oocytes and embryos. Therefore, the aim of this work was to analyze the leptin protein distribution within bovine oocytes and preimplantation embryos matured and fertilized (in vitro). The material for this work consisted of oocytes collected from slaughterhouse ovaries and sperm collected from AI bulls. In vitro oocyte maturation and fertilization were carried out according to the method described by Makarevich and Markkula (2002 Biol. Reprod. 66, 386–392). The preliminary experiment of leptin protein localization by immunofluorescent staining included immature and matured oocytes and blastocysts. Oocytes and embryos were fixed in PBS containing 4% paraformaldehyde and reacted with affinity-purified polyclonal rabbit primary antibody directed against leptin (0.1 mg/mL; Ob (Y20), Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); and then exposed to secondary goat-anti-rabbit antibody (1.0 mg/mL; Santa Cruz Biotechnology Inc.)-fluorescein isothiocyanate (FITC) conjugate. Finally, chromatin was visualized by propidium iodide staining (0.5 μg/mL). Slides were examined under a conventional fluorescence microscope (Nikon) and confocal microscope (Zeiss). The preliminary results demonstrate that the distribution of leptin differed between immature and mature oocytes: it was spherical in immature oocytes (a rim beneath the oolemma) whereas it became evenly distributed after maturation. In blastocysts, leptin signals were present inboth the trophoblast cells and in the ICM cells. This is in contrast with studies on mouse embryos which showed the presence of the LEP protein in the trophoblast only. Future experiments will include studies of embryos at the 2-cell, 4-cell, 8–16-cell, and morula stages. The present study for the first time shows the pattern of leptin protein distribution within bovine oocytes and preattachment embryos.

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