Interaction of epiblast and hypoblast in the formation of the primitive streak and the embryonic axis in chick, as revealed by hypoblast-rotation experiments

Development ◽  
1981 ◽  
Vol 61 (1) ◽  
pp. 133-144
Author(s):  
Yehudit Azar ◽  
Hefzibah Eyal-Giladi

Three types of experiments were performed to determine the interaction between the epiblast and hypoblast for primitive streak formation: (1) Hypoblasts of blastoderms from stages XIII E.G & K to 3 H & H were separated from the epiblasts and rotated by 90° counterclockwise; (2) hypoblasts from stages XIII E.G & K to 3 H & H blastoderms were rotated by 180° (3) hypoblasts were exchanged between blastoderms of different developmental stages and placed at 90° counterclockwise to the axis of the recipient epiblast. In all blastoderms studied only a single PS developed. After rotation of the hypoblast by 90°, the direction of the PS was according to the orientation of the hypoblast at stage XIII, whereas at older stages it gradually shifted towards the axis of the epiblast. At stage 3− H & H the PS is already imprinted in the epiblast and cannot be shifted. After rotation of the hypoblast by 180° the PS originated at the point near the marginal zone at which the inductive part of the hypoblast interacted with a competent epiblast. Conclusions are drawn about the dynamics of the inductiveness of the hypoblast and the competence of the epiblast for the PS formation and orientation.

Development ◽  
1984 ◽  
Vol 82 (1) ◽  
pp. 253-266
Author(s):  
P. P. L. Tam

The caudal end of the embryonic axis consists of the primitive streak and the tail bud. Small fragments of this caudal tissue were transplanted from mouse embryos of various developmental stages to the kidney capsule in order to test their histogenetic capacity. The variety of mature tissues obtained from these small fragments was similar to that obtained by grafting a larger caudal portion of the embryo. Initially, the grafted tissue broke up into loose masses of embryonic mesenchyme and this was later re-organized into more compact tissues and into cysts that were lined with various types of epithelia. After 14 days in the ectopic site, grafted tissues coming from embryos of the primitive-streak, the early-somite and the forelimb-bud stages differentiated into structures that has presumably originated from the three embryonic germ layers. Many of these structures were related to the caudal region of the adult body, such as the mid- and hindgut segments and urogenital derivatives. The histogenetic capacity for endodermal tissues and urogenital organs was lost when the grafted tissue consisted entirely of the tail bud of the hindlimb-bud-stage embryos. The behaviour of the caudal tissues suggested that (1) the primordia for the various parts of embryonic body were derived from a small progenitor population in the primitive streak and the tail bud, and (2) the histogenetic capacity of this population changed during development.


Development ◽  
2001 ◽  
Vol 128 (15) ◽  
pp. 2915-2927 ◽  
Author(s):  
Isaac Skromne ◽  
Claudio D. Stern

The posterior marginal zone (PMZ) of the chick embryo has Nieuwkoop centre-like properties: when transplanted to another part of the marginal zone, it induces a complete embryonic axis, without making a cellular contribution to the induced structures. However, when the PMZ is removed, the embryo can initiate axis formation from another part of the remaining marginal zone. Chick Vg1 can mimic the axis-inducing ability of the PMZ, but only when misexpressed somewhere within the marginal zone. We have investigated the properties that define the marginal zone as a distinct region. We show that the competence of the marginal zone to initiate ectopic primitive streak formation in response to cVg1 is dependent on Wnt activity. First, within the Wnt family, only Wnt8C is expressed in the marginal zone, in a gradient decreasing from posterior to anterior. Second, misexpression of Wnt1 in the area pellucida enables this region to form a primitive streak in response to cVg1. Third, the Wnt antagonists Crescent and Dkk-1 block the primitive streak-inducing ability of cVg1 in the marginal zone. These findings suggest that Wnt activity defines the marginal zone and allows cVg1 to induce an axis. We also present data suggesting some additional complexity: first, the Vg1 and Wnt pathways appear to regulate the expression of downstream components of each other’s pathway; and second, misexpression of different Wnt antagonists suggests that different classes of Wnts may cooperate with each other to regulate axis formation in the normal embryo.


Development ◽  
1992 ◽  
Vol 116 (3) ◽  
pp. 819-830 ◽  
Author(s):  
H. Eyal-Giladi ◽  
A. Debby ◽  
N. Harel

Posterior marginal zone sections with or without Koller's sickle were cut out of stage X, XI and XII E.G&K blastoderms, labelled with the fluorescent dye rhodamine-dextran-lysine (RDL) and returned to their original location. In control experiments, a similar lateral section of the marginal zone was identically treated. Different blastoderms were incubated at 37°C for different periods and were fixed after reaching stages from XII E.G&K to 4 H&H. The conclusions drawn from the analysis of the distribution pattern of the labelled cells in the serially sectioned blastoderms concern the cellular contributions to both the forming hypoblast and the forming primitive streak. Koller's sickle and the marginal zone behind it were found to contribute all the centrally located cells of the growing hypoblast. The lengthening pregastrulation PS (until stage 3+ H&H) was found to be entirely composed of epiblastic cells that at stage X were located in a narrow strip anterior to Koller's sickle. A model is proposed to integrate the results spatially and temporally.


Development ◽  
1997 ◽  
Vol 124 (24) ◽  
pp. 5127-5138 ◽  
Author(s):  
S.B. Shah ◽  
I. Skromne ◽  
C.R. Hume ◽  
D.S. Kessler ◽  
K.J. Lee ◽  
...  

In the chick embryo, the primitive streak is the first axial structure to develop. The initiation of primitive streak formation in the posterior area pellucida is influenced by the adjacent posterior marginal zone (PMZ). We show here that chick Vg1 (cVg1), a member of the TGFbeta family of signalling molecules whose homolog in Xenopus is implicated in mesoderm induction, is expressed in the PMZ of prestreak embryos. Ectopic expression of cVg1 protein in the marginal zone chick blastoderms directs the formation of a secondary primitive streak, which subsequently develops into an ectopic embryo. We have used cell marking techniques to show that cells that contribute to the ectopic primitive streak change fate, acquiring two distinct properties of primitive streak cells, defined by gene expression and cell movements. Furthermore, naive epiblast explants exposed to cVg1 protein in vitro acquire axial mesodermal properties. Together, these results show that cVg1 can mediate ectopic axis formation in the chick by inducing new cell fates and they permit the analysis of distinct events that occur during primitive streak formation.


Development ◽  
1994 ◽  
Vol 120 (9) ◽  
pp. 2501-2509 ◽  
Author(s):  
H. Eyal-Giladi ◽  
T. Lotan ◽  
T. Levin ◽  
O. Avner ◽  
J. Hochman

Hypoblast cells of posterior marginal zone origin have been shown previously to be the inducers of primitive streak in the avian embryo. Here we checked: (1) whether the above cells acquire their inductivity while still whithin the marginal zone; (2) can inductivity be found in supernatants of defined blastodermic regions; (3) can differences in the electrophoretic pattern be shown between inducing and non-inducing tissue fragments and their conditioned media, which might give a clue as to what the inductive substance is. The following observations were made: 1. (a) Stage X chick posterior marginal zone cells prior to their migration into the hypoblast do not induce a primitive streak, when applied to a stage XIII competent epiblast central disc. (b) A posterior marginal zone fragment, when applied to an epiblast central disc, even after being preincubated for up to 9 hours in vitro, is still non-inductive. (c) Mechanically fragmented stage X posterior marginal zones when applied as a layer to epiblast central discs are non-inductive. (d) Hypoblastic tissue in strip form induces a primitive streak. 2. Competent stage XIII epiblast central discs (chick) were incubated for 2 hours in supernatants of stage XIII epiblasts or hypoblasts. Whereas no inductive effect was exerted by the epiblast supernatant, primitive streaks developed in about 50% of the epiblast central discs incubated in the hypoblast supernatant. 3. Electrophoretic analysis (quails) reveals a protein of 28x10-3 Mr that is enriched in both hypoblastic tissue and its incubation medium and not in the epiblast + marginal zone + area opaca and their incubation medium. These findings suggest a possible correlation between this protein and the induction process.


Development ◽  
1998 ◽  
Vol 125 (17) ◽  
pp. 3521-3534 ◽  
Author(s):  
R.F. Bachvarova ◽  
I. Skromne ◽  
C.D. Stern

In the preprimitive streak chick embryo, the search for a region capable of inducing the organizer, equivalent to the Nieuwkoop Center of the amphibian embryo, has focused on Koller's sickle, the hypoblast and the posterior marginal zone. However, no clear evidence for induction of an organizer without contribution from the inducing tissue has been provided for any of these structures. We have used DiI/DiO labeling to establish the fate of midline cells in and around Koller's sickle in the normal embryo. In the epiblast, the boundary between cells that contribute to the streak and those that do not lies at the posterior edge of Koller's sickle, except at stage X when it lies slightly more posteriorly in the epiblast. Hypoblast and endoblast (a second lower layer formed under the streak) have distinct origins in the lower layer, and goosecoid expression distinguishes between them. We then used anterior halves of chick prestreak embryos as recipients for grafts of quail posterior marginal zone; quail cells can be identified subsequently with a quail-specific antibody. Anterior halves alone usually formed a streak, most often from the posterior edge. Quail posterior marginal zones without Koller's sickle were grafted to the anterior side of anterior halves. These grafts were able to increase significantly the frequency of streaks arising from the anterior pole of stage X-XI anterior halves without contributing to the streak or node. Stage XII anterior halves no longer responded. A goosecoid-expressing hypoblast did not form under the induced streak, indicating that it is not required for streak formation. We conclude that the marginal zone posterior to Koller's sickle can induce a streak and node, without contributing cells to the induced streak.


Development ◽  
1990 ◽  
Vol 109 (3) ◽  
pp. 667-682 ◽  
Author(s):  
C.D. Stern

The marginal zone of the chick embryo has been shown to play an important role in the formation of the hypoblast and of the primitive streak. In this study, time-lapse filming, fate mapping, ablation and transplantation experiments were combined to study its contribution to these structures. It was found that the deep (endodermal) portion of the posterior marginal zone contributes to the hypoblast and to the junctional endoblast, while the epiblast portion of the same region contributes to the epiblast of the primitive streak and to the definitive (gut) endoderm derived from it. Within the deep part of the posterior marginal zone, a subpopulation of HNK-1-positive cells contributes to the hypoblast. Removal of the deep part of the marginal zone prevents regeneration of the hypoblast but not the formation of a primitive streak. Removal of both layers of the marginal zone leads to a primitive streak of abnormal morphology but mesendodermal cells nevertheless differentiate. These results show that the two main properties of the posterior marginal zone (contributing to the hypoblast and controlling the site of primitive streak formation) are separable, and reside in different germ layers. This conclusion does not support the idea that the influence of the posterior marginal zone on the development of axial structures is due to it being the source of secondary hypoblast cells.


Development ◽  
1993 ◽  
Vol 119 (4) ◽  
pp. 1147-1160 ◽  
Author(s):  
C.R. Hume ◽  
J. Dodd

To begin to examine the possibility that Wnt proteins act as cell signalling molecules during chick embryogenesis, PCR was used to identify Wnt genes expressed in Hensen's node. We have identified a novel member of the Wnt gene family, Cwnt-8C, which is expressed prior to gastrulation in the posterior marginal zone, the primitive streak and Hensen's node. Injection of Cwnt-8C mRNA into Xenopus embryos caused axis duplication and dorsalization of mesodermal tissues. During neurulation, Cwnt-8C is expressed transiently in a restricted domain of the prospective hindbrain neurectoderm that will give rise to rhombomere 4. This domain is defined prior to the formation of rhombomere boundaries and also precedes the up-regulation and restriction of expression of Hox B1 in the same region. Thus, Cwnt-8C is potentially involved in the regulation of axis formation and hindbrain patterning.


In chick blastoderms at primitive streak stage, lengths of the primitive streak were cut out and replaced with their antero-posterior orientation reversed. In some experiments the region immediately in front of the primitive streak (presumptive prechordal head) was also included in the excisedpiece. Control operations involving excision and replacement without reversal were also performed. The embryos were subsequently grown in vitro by Waddington’s technique. After reversal of a variety of different parts of the streak at various developmental stages, many cases of regulative development were obtained. In these, the original orientation of the blastoderm was maintained, and while there were abnormalities of various kinds in the embryos, they were no different from the abnormalities found in the controls. Very occasionally the regulated axis was partially doubled after a reversal, though not after a control operation. A few specimens which had undergone reversal of long pieces of the primitive streak and had completely healed showed a failure of regulation in that there was some tendency for the reversed-piece to develop according to its own orientation. But at best this reversed differentiation was very distorted and incomplete. Evidently the orientation of the primitive streak does not at any stage control the orientation of the embryo; and the primitive streak, when it is fully developed and contains most of the presumptive axial material, is highly labile in its powers of differentiation. In spite of its well-known ‘organizer' activity, the primitive streak is subject to control by the surrounding blastoderm.


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