scholarly journals Dorsalization and neural induction: properties of the organizer in Xenopus laevis

Development ◽  
1983 ◽  
Vol 78 (1) ◽  
pp. 299-317 ◽  
Author(s):  
J. C. Smith ◽  
J. M. W. Slack
Keyword(s):  
Horseradish Peroxidase ◽  
Xenopus Laevis ◽  
Neural Tube ◽  
Marginal Zone ◽  
Neural Induction ◽  
Traditional View ◽  
Posterior Half ◽  
Secondary Embryo ◽  
Graft Tissue ◽  
Host Embryo

We have studied the action of the organizer in Xenopus laevis using grafts labelled with horseradish peroxidase (HRP). Orthotopic grafts of the dorsal marginal zone (the organizer) from an HRP-labelled embryo into an unlabelled host showed that this region contributes to the anterior archenteron wall, to the entire craniocaudal extent of the notochord and to a few cells in the somites. Little or no contribution was made to the neural tube. Orthotopic grafts of the ventral marginal zone (the tissue that responds to a grafted organizer) indicated that it only contributes to the posterior half of the embryo. Within this region it spreads around the entire ventrolateral mesoderm, occasionally contributing a few cells to the somites. The posterior endoderm was also heavily labelled. When the dorsal marginal zone from an HRP-labelled embryo was inserted into a slit cut in the ventral marginal zone of an unlabelled host a mirror-symmetrical double-dorsal duplicated embryo resulted, in which only the notochord and a few cells in the somites of the secondary embryo were derived from the graft. The bulk of the secondary somites was, therefore, derived from host ventral marginal zone tissue which normally makes very little contribution to the somites. This indicates that host ventral marginal zone becomes dorsalized by the graft. The neural tube of the secondary embryo was also unlabelled, showing that it was induced by the influence of the graft on the overlying ectoderm, which normally forms ventral epidermis. We have also grafted ventral marginal zone tissue into a slit cut into the dorsal marginal zone of a host embryo. HRP-labelled tissue was grafted into an unlabelled embryo and vice versa. This graft did not produce a double ventral embryo and this reinforces the traditional view that the dorsal marginal zone is a special signalling region. Instead, the resulting embryos usually had a twinned notochord with the graft tissue in between, differentiated as somite. This confirms that juxtaposing ventral and dorsal marginal zone ‘dorsalizes’ the ventral tissue but does not affect the dorsal tissue which differentiates, as usual, as notochord. Thus, our results allow us to conclude that the organizer mediates two distinct interactions in bringing about the formation of duplicated embryos. The first is dorsalization of adjacent ventral mesoderm and the second is the induction of neuroepithelium from ectoderm overlying the new archenteron roof.

Competition during innervation of embryonic amphibian head skin

1983 ◽  
Vol 218 (1210) ◽  
pp. 49-59 ◽  
Keyword(s):  
Horseradish Peroxidase ◽  
Xenopus Laevis ◽  
Trigeminal Ganglion ◽  
Nerve Endings ◽  
Movement Detector ◽  
Dorsal Midline ◽  
Intact Side ◽  
Free Nerve Endings ◽  
Peripheral Innervation ◽  
Peripheral Sensory

We have examined the initial innervation of the head skin in Xenopus laevis embryos which is by two classes of trigeminal mechanoreceptor with beaded ‘free’ nerve-endings. By recording receptive areas electrophysiologically and staining peripheral sensory neurites with horseradish peroxidase, we have shown that ‘movement detector’ neurites from one trigeminal ganglion do not normally cross the dorsal midline of the head to innervate areas of skin on the opposite side. However, if one trigeminal ganglion is removed before peripheral innervation starts, movement detector neurites from the intact side will now cross the midline to innervate contralateral skin. These observations suggest a specific competitive interaction between movement detector neurites during their innervation of head skin. The second class of receptor, ‘rapid transient’ detectors, have a different pattern of innervation, crossing the midline in both normal and operated animals.

Neural induction and regionalisation in the chick embryo

Development ◽  
1992 ◽  
Vol 114 (3) ◽  
pp. 729-741 ◽  
Author(s):  
K.G. Storey ◽  
J.M. Crossley ◽  
E.M. De Robertis ◽  
W.E. Norris ◽  
C.D. Stern
Keyword(s):  
Spinal Cord ◽  
Nervous System ◽  
Molecular Markers ◽  
Chick Embryo ◽  
Normal Development ◽  
Neural Induction ◽  
Stage 4 ◽  
Embryo Chick ◽  
Host Embryo ◽  
Embryonic Region

Induction and regionalisation of the chick nervous system were investigated by transplanting Hensen's node into the extra-embryonic region (area opaca margin) of a host embryo. Chick/quail chimaeras were used to determine the contributions of host and donor tissue to the supernumerary axis, and three molecular markers, Engrailed, neurofilaments (antibody 3A10) and XlHbox1/Hox3.3 were used to aid the identification of particular regions of the ectopic axis. We find that the age of the node determines the regions of the nervous system that form: young nodes (stages 2–4) induced both anterior and posterior nervous system, while older nodes (stages 5–6) have reduced inducing ability and generate only posterior nervous system. By varying the age of the host embryo, we show that the competence of the epiblast to respond to neural induction declines after stage 4. We conclude that during normal development, the initial steps of neural induction take place before stage 4 and that anteroposterior regionalisation of the nervous system may be a later process, perhaps associated with the differentiating notochord. We also speculate that the mechanisms responsible for induction of head CNS differ from those that generate the spinal cord: the trunk CNS could arise by homeogenetic induction by anterior CNS or by elongation of neural primordia that are induced very early.

Developmental potential of neural crest-derived cells migrating from segments of developing quail bowel back-grafted into younger chick host embryos

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 411-423 ◽  
Author(s):  
T.P. Rothman ◽  
N.M. Le Douarin ◽  
J.C. Fontaine-Perus ◽  
M.D. Gershon
Keyword(s):  
Neural Crest ◽  
Neural Tube ◽  
Peripheral Nerves ◽  
Sympathetic Ganglia ◽  
Limb Bud ◽  
Pigment Cells ◽  
Developmental Potential ◽  
Migratory Experience ◽  
Host Embryo

The technique of back-transplantation was used to investigate the developmental potential of neural crest-derived cells that have migrated to and colonized the avian bowel. Segments of quail bowel (removed at E4) were grafted between the somites and neural tube of younger (E2) chick host embryos. Grafts were placed at a truncal level, adjacent to somites 14–24. Initial experiments, done in vitro, confirmed that crest-derived cells are capable of migrating out of segments of foregut explanted at E4. The foregut, which at E4 has been colonized by cells derived from the vagal crest, served as the donor tissue. Comparative observations were made following grafts of control tissues, which included hindgut, lung primordia, mesonephros and limb bud. Additional experiments were done with chimeric bowel in which only the crest-derived cells were of quail origin. Targets in the host embryos colonized by crest-derived cells from the foregut grafts included the neural tube, spinal roots and ganglia, peripheral nerves, sympathetic ganglia and the adrenals, but not the gut. Donor cells in these target organs were immunostained by the monoclonal antibody, NC-1, indicating that they were crest-derived and developing along neural or glial lineages. Some of the crest-derived cells (NC-1-immunoreactive) that left the bowel and reached sympathetic ganglia, but not peripheral nerves or dorsal root ganglia, co-expressed tyrosine hydroxylase immunoreactivity, a neural characteristic never expressed by crest-derived cells in the avian gut. None of the cells leaving enteric back-grafts produced pigment. Cells of mesodermal origin were also found to leave donor explants and aggregate in dermis and feather germs near the grafts. These observations indicate that crest-derived cells, having previously migrated to the bowel, retain the ability to migrate to distant sites in a younger embryo. The routes taken by these cells appear to reflect, not their previous migratory experience, but the level of the host embryo into which the graft is placed. Some of the population of crest-derived cells that leave the back-transplanted gut remain capable of expressing phenotypes that they do not express within the bowel in situ, but which are appropriate for the site in the host embryo to which they migrate.

Expression of N-CAM precedes neural induction in Pleurodeles waltl (urodele, amphibian)

Development ◽  
1989 ◽  
Vol 106 (4) ◽  
pp. 675-683 ◽  
Author(s):  
J.P. Saint-Jeannet ◽  
F. Foulquier ◽  
C. Goridis ◽  
A.M. Duprat
Keyword(s):  
Monoclonal Antibody ◽  
Nervous System ◽  
Xenopus Laevis ◽  
Neural Induction ◽  
Gastrula Stage ◽  
Pleurodeles Waltl ◽  
Early Gastrula

The appearance and localization of N-CAM during neural induction were studied in Pleurodeles waltl embryos and compared with recent contradictory results reported in Xenopus laevis. A monoclonal antibody raised against mouse N-CAM was used. In the nervous system of Pleurodeles, it recognized two glycoproteins of 180 and 140×10(3) M(r) which are the Pleurodeles equivalent of N-CAM-180 and -140. Using this probe for immunohistochemistry and immunocytochemistry, we showed that N-CAM was already expressed in presumptive ectoderm at the early gastrula stage. In late gastrula embryos, a slight increase in staining was observed in the neurectoderm, whereas the labelling persisted in the noninduced ectoderm. When induced ectodermal cells were isolated at the late gastrula stage and cultured in vitro up to 14 days, a faint polarized labelling of cells was observed initially. During differentiation, the staining increased and became progressively restricted to differentiating neurons.

The fate of cells in the tailbud of Xenopus laevis

Development ◽  
2000 ◽  
Vol 127 (2) ◽  
pp. 255-267 ◽  
Author(s):  
R.L. Davis ◽  
M.W. Kirschner
Keyword(s):  
Xenopus Laevis ◽  
Small Groups ◽  
Neural Tube ◽  
Cell Types ◽  
Similar Distribution ◽  
Germ Layer ◽  
Cell Determination ◽  
Multipotent Cells ◽  
Axial Development

The vertebrate tailbud and trunk form very similar tissues. It has been a controversial question for decades whether cell determination in the developing tail proceeds as part of early axial development or whether it proceeds by a different mechanism. To examine this question more closely, we have used photoactivation of fluorescence to mark small neighborhoods of cells in the developing tailbud of Xenopus laevis. We show that, in one region of the tailbud, very small groups of adjacent cells can contribute progeny to the neural tube, notochord and somitic muscle, as well as other identified cell types within a single embryo. Groups averaging three adjacent cells at a later stage can contribute progeny with a similar distribution. Our data suggest that the tailbud contains multipotent cells that make very late germ-layer decisions.

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.

Mesoderm-inducing factors and Spemann's organiser phenomenon in amphibian development

Development ◽  
1989 ◽  
Vol 107 (2) ◽  
pp. 229-241 ◽  
Author(s):  
J. Cooke
Keyword(s):  
Growth Factor ◽  
Pattern Formation ◽  
Marginal Zone ◽  
Natural Control ◽  
Graft Size ◽  
Two Factors ◽  
Inducing Factors ◽  
Mesoderm Inducing Factors ◽  
Host Embryo ◽  
Factor Concentration

Certain proteins from ‘growth factor’ families can initiate mesodermal development in animal cap cells of the amphibian blastula. Cells that are in early stages of their response to one such factor, XTC-MIF (Smith et al. 1988), initiate the formation of a new axial body plan when grafted to the ventral marginal zone of a similarly aged host embryo (Cooke et al. 1987). This replicates the natural control of this phase of development by the dorsal blastoporal lip when similarly grafted; the classical ‘organiser’ phenomenon. I have explored systematically the effect, upon the outcome of this pattern formation using defined inducing factors, of varying graft size, XTC-MIF concentration to which graft cells were exposed, length of exposure before grafting, and host age. The ‘mesodermal organiser’ status, evoked by the factor, appears to be stable, and the variables most influencing the degree of completeness and orderliness of second patterns are graft size and factor concentration. Inappropriately large grafts are not effective. A Xenopus basic fibroblast growth factor homologue, present in the embryo and known to be a strong inducer but of mesoderm with a different character from that induced by XTC-MIF, produced no episode of pattern formation at all when tested in the procedure described in this paper. Organiser status of grafts that have been exposed to mixtures of the two factors is set entirely by the supplied XTC-MIF concentration. Lineage labelling of these grafts, and of classical dorsal lip grafts, reveals closely similar though not identical patterns of contribution to the new structure within the host. Implications of the results for the normal mechanism of body pattern formation are discussed.

Regional specification within the mesoderm of early embryos of Xenopus laevis

Development ◽  
1987 ◽  
Vol 100 (2) ◽  
pp. 279-295 ◽  
Author(s):  
L. Dale ◽  
J.M. Slack
Keyword(s):  
Xenopus Laevis ◽  
Marginal Zone ◽  
Mesoderm Induction ◽  
Intermediate Type ◽  
Cell Stage ◽  
Early Embryos ◽  
Significant Difference ◽  
Cell Embryo ◽  
Single Blastomeres

We have further analysed the roles of mesoderm induction and dorsalization in the formation of a regionally specified mesoderm in early embryos of Xenopus laevis. First, we have examined the regional specificity of mesoderm induction by isolating single blastomeres from the vegetalmost tier of the 32-cell embryo and combining each with a lineage-labelled (FDA) animal blastomere tier. Whereas dorsovegetal (D1) blastomeres induce ‘dorsal-type’ mesoderm (notochord and muscle), laterovegetal and ventrovegetal blastomeres (D2–4) induce either ‘intermediate-type’ (muscle, mesothelium, mesenchyme and blood) or ‘ventral-type’ (mesothelium, mesenchyme and blood) mesoderm. No significant difference in inductive specificity between blastomeres D2, 3 and 4 could be detected. We also show that laterovegetal and ventrovegetal blastomeres from early cleavage stages can have a dorsal inductive potency partially activated by operative procedures, resulting in the induction of intermediate-type mesoderm. Second, we have determined the state of specification of ventral blastomeres by isolating and culturing them in vitro between the 4-cell stage and the early gastrula stage. The majority of isolates from the ventral half of the embryo gave extreme ventral types of differentiation at all stages tested. Although a minority of cases formed intermediate-type and dorsal-type mesoderms we believe these to result from either errors in our assessment of the prospective DV axis or from an enhancement, provoked by microsurgery, of some dorsal inductive specificity. The results of induction and isolation experiments suggest that only two states of specification exist in the mesoderm of the pregastrula embryo, a dorsal type and a ventral type. Finally we have made a comprehensive series of combinations between different regions of the marginal zone using FDA to distinguish the components. We show that, in combination with dorsal-type mesoderm, ventral-type mesoderm becomes dorsalized to the level of intermediate-type mesoderm. Dorsal-type mesoderm is not ventralized in these combinations. Dorsalizing activity is confined to a restricted sector of the dorsal marginal zone, it is wider than the prospective notochord and seems to be graded from a high point at the dorsal midline. The results of these experiments strengthen the case for the three-signal model proposed previously, i.e. dorsal and ventral mesoderm inductions followed by dorsalization, as the simplest explanation capable of accounting for regional specification within the mesoderm of early Xenopus embryos.

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