Studies on self-differentiating and induction capacities of Hensen's node using intracoelomic grafting technique

Development ◽  
1972 ◽  
Vol 28 (3) ◽  
pp. 547-558
Author(s):  
J. R. Viswanath ◽  
Leela Mulherkar

Living Hensen's node of the definitive primitive streak of chick embryo was prepared into ‘sandwiches’ with the competent ectoderm and the sandwich grafts were transplated into the 2·5 day chick embryo using the intracoelomic grafting technique of Hamburger. One hundred and twenty-four grafts were prepared and transplanted intracoelomically, 28 grafts were lost due to the death of the host embryos, 63 grafts did not differentiate at all, but 33 well-defined grafts were recovered, after cultivating the transplanted hosts for 12–14 days. All kinds of tissues from feather germs to neural tissue were found to have differentiated in the grafts. The more frequently occurring tissues were feather germs, epidermal vesicle, neural tissue, kidney and muscle. Other differentiations were the cartilage notochord and gut. No definite combination pattern has emerged from the tissues. But when the tissues were traced to their germ-layer derivation, 22 of them belonged to the mesodermal complex, 11 to the ectodermal complex and 8 to the endodermal complex. In the light of the above results, the probable existence of a mesodermal factor and an ectodermal factor independently responsible for the respective differentiations, as also the competence of the ectoderm, is discussed.

Development ◽  
1998 ◽  
Vol 125 (3) ◽  
pp. 473-484 ◽  
Author(s):  
K.G. Storey ◽  
A. Goriely ◽  
C.M. Sargent ◽  
J.M. Brown ◽  
H.D. Burns ◽  
...  

Signals that induce neural cell fate in amniote embryos emanate from a unique cell population found at the anterior end of the primitive streak. Cells in this region express a number of fibroblast growth factors (FGFs), a group of secreted proteins implicated in the induction and patterning of neural tissue in the amphibian embryo. Here we exploit the large size and accessibility of the early chick embryo to analyse the function of FGF signalling specifically during neural induction. Our results demonstrate that extraembryonic epiblast cells previously shown to be responsive to endogenous neural-inducing signals express early posterior neural genes in response to local, physiological levels of FGF signal. This neural tissue does not express anterior neural markers or undergo neuronal differentiation and forms in the absence of axial mesoderm. Prospective mesodermal tissue is, however, induced and we present evidence for both the direct and indirect action of FGFs on prospective posterior neural tissue. These findings suggest that FGF signalling underlies a specific aspect of neural induction, the initiation of the programme that leads to the generation of the posterior central nervous system.


Development ◽  
1993 ◽  
Vol 118 (2) ◽  
pp. 389-399 ◽  
Author(s):  
E.J. Sanders ◽  
M. Varedi ◽  
A.S. French

Cell proliferation in the gastrulating chick embryo was assessed using two independent techniques which mark cells in S phase of the mitotic cycle: nuclear incorporation of bromodeoxyuridine (BrdU) detected immunocytochemically and immunolocalization of proliferating cell nuclear antigen (PCNA). Computer-reconstructed maps were produced showing the distribution of labelled nuclei in the primitive streak and the cell layers. These distributions were also normalized to take into account regional differences in cell density across the embryo. Results from a 2 hour pulse of BrdU indicated that although cells at caudal levels of the primitive streak showed the highest incorporation, this region showed a similar proportion of labelled cells to the surrounding caudal regions of the epiblast and mesoderm when normalized for cell density. The entire caudal third of the embryo showed the highest proportion of cells in S phase. Cells of Hensen's node showed a relatively low rate of incorporation and, although the chordamesoderm cells showed many labelled nuclei, this appeared to be a reflection of a high cell density in this region. Combining this result with results from a 4 hour pulse of BrdU permitted mapping of cell generation time across the entire embryo. Generation times ranged from a low value of approximately 2 hours at caudal levels of both the epiblast and mesoderm, to an upper value of approximately 10 hours in the rostral regions of the primitive streak, in the mid-lateral levels of the epiblast and in the chordamesoderm rostral to Hensen's node. Cells at caudal regions of the primitive streak showed a generation time of approximately 5 hours. Taking into account that cells are generally considered to be continuously moving through the primitive streak, we conclude that cell division, as judged by generation time, is greatly reduced during transit through this region, despite the presence there of cells in S phase and M phase. Immunocytochemical localization of PCNA-positive nuclei gave generally similar distributions to those obtained with BrdU incorporation, confirming that this endogenous molecule is a useful S-phase marker during early embryogenesis. Mid-levels and caudal levels of the primitive streak showed the highest numbers of positive nuclei, and the highest proportion of labelling after cell density was accounted for. As with BrdU incorporation, the highest proportions of PCNA-positive nuclei were found towards the caudal regions of the epiblast and mesoderm. These results suggest that the differential growth of the caudal region of the embryo at this time is a direct consequence of elevated levels of cell proliferation in this region.(ABSTRACT TRUNCATED AT 400 WORDS)


Development ◽  
1968 ◽  
Vol 20 (3) ◽  
pp. 247-260
Author(s):  
Teresa Rogulska

Suggestive evidence for the extragonadal origin of germ cells in birds was first presented by Swift (1914), who described primordial germ cells in the chick embryo at as early a stage as the primitive streak. According to Swift, primordial germ cells are originally located extra-embryonically in the anterior part of the blastoderm and occupy a crescent-shaped region (‘germinal crescent’) on the boundary between area opaca and area pellucida. Swift also found that primordial germ cells later enter into the blood vessels, circulate together with the blood throughout the whole blastoderm and finally penetrate into the genital ridges, where they become definitive germ cells. Swift's views have been confirmed in numerous descriptive and experimental investigations. Among the latter, the publications of Willier (1937), Simon (1960) and Dubois (1964a, b, 1965a, b, 1966) merit special attention. Dubois finally proved that the genital ridges exert a strong chemotactic influence on the primordial germ cells.


Development ◽  
1986 ◽  
Vol 95 (1) ◽  
pp. 81-93
Author(s):  
E. J. Sanders

Mesodermal tissue from the chick embryo at various stages of early differentiation was cultured in hydrated gels of type I collagen in the presence and absence of transferrin. Primary mesoderm explants from primitive-streak-stage embryos responded to the presence of avian transferrin by significantly improved outgrowth which appeared to be related to the ability of the cells to attach to, and migrate in, the collagen. No evidence was obtained which suggested that this observation was dependent on increased cell proliferation. This outgrowth enhancement was not duplicated by transferrin of human origin. The avian transferrin did not produce this effect on cells cultured on plastic substrata, suggesting that the species-specific effect involves modulation by the extracellular matrix. Mesoderm explants from somite stages of development showed no increase in outgrowth in the presence of either avian or human transferrin as judged by counting the number of outwandering cells. Ultrastructural immunocytochemistry indicated surface binding of transferrin by cells in the gels, and the presence of endogenous transferrin on the surfaces of mesoderm cells in situ and in their extracellular environment. It is suggested that by binding to cell surface receptors, transferrin may be able to influence the strength of cellular adhesion to collagen and hence the capacity for cell locomotion.


1937 ◽  
Vol 14 (3) ◽  
pp. 319-334 ◽  
Author(s):  
M. ABERCROMBIE ◽  
C. H. WADDINGTON

1. Grafts consisting of pieces of primitive streak from blastoderms in the primitive streak stage were placed under the primitive streak of blastoderms also in this stage. 2. Various effects of the host on the graft are described, particularly the reversal of the antero-posterior orientation of the graft, the alteration of the regional character of the graft so as to conform with the host tissues at the same level, the suppression of differentiation in the posterior end of the primitive streak, and the incorporation of the graft tissues into the host. 3. A considerable number of inductions occurred, since the host axis often apparently shifts to one side of the graft. The inductions are of two kinds, the normal evocation by graft mesoderm, resulting usually in the formation of superfluous neural tissue; and the complementary induction of a normal secondary axis, which it is supposed is most often due to the preliminary induction of a primitive streak in the host. 4. Various effects of the graft on the host occur. In particular the disturbance of the head mesenchyme suggests that foregut diverticula are produced where head mesenchyme joins lateral plate mesothelium.


1936 ◽  
Vol 13 (2) ◽  
pp. 219-236
Author(s):  
C. H. WADDINGTON ◽  
A. COHEN

1. Experiments were made on the development of the head of chicken embryos cultivated in vitro. 2. Defects in the presumptive head region of primitive streak embryos are regulated completely if the wound fills up before the histogenesis of neural tissue begins in the head-process stage. Different methods by which the hole is filled are described. 3. No repair occurs in the head-process and head-fold stages, and in this period two masses of neural tissue cannot heal together. 4. Median defects, even if repaired as regards neural tissue, cause a failure of the ventral closure of the foregut. The lateral evaginations of the gut develop typically in atypical situations. The headfold may break through and join up with the endoderm in such a way that the gut acquires an anterior opening. 5. The paired heart rudiments may develop separately. The separate vesicles begin to contract at a time appropriate to the development of the embryo as a whole. The two hearts are mirror images, the left one having the normal curvature, but the embryos do not survive long enough for the hearts to acquire a very definite shape. 6. The forebrain has a considerable capacity for repair in the early somite stages (five to twenty-five somites). One-half of the forebrain can remodel itself into a complete forebrain. In some cases the neural plate and epidermis grow together over the wound, in others the epidermis and mesenchyme make the first covering, leaving a space along the inside of which the neural tissue grows. The neural tissue may become a very thin sheet. 7. The repaired forebrain may induce the formation of a nasal placode from the non-presumptive nasal epidermis which covers the wound. 8. If the optic vesicle is entirely removed, a new one is not formed, but parts of the vesicle can regulate to complete eye-cups, either when still attached to the forebrain or after being isolated in the extra-embryonic regions of another embryo. 9. Injured optic vesicles induce lenses from the non-presumptive epidermis which grows over the wound. Transplanted optic neural tissue from embryos of about five somites induces the formation of lentoids from extra-embryonic ectoderm, but only in a small proportion of cases. 10. The presumptive lens epidermis can produce a slight thickening even when contact with the optic cup is prevented. 11. The significance of periods of minimum regulatory power for the concept of determination is discussed. 12. The data concerning lens formation are discussed in terms of the field concept.


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