Determination of anteroposterior polarity in the axolotl forelimb by an interaction between limb and flank rudiments

Development ◽  
1977 ◽  
Vol 39 (1) ◽  
pp. 151-168
Author(s):  
J. M. W. Slack

1. It is shown that the mesoderm in the prospective forelimb-bud of the axolotl embryo is thickened and divided into somatic and splanchnic layers, while that of the flank is thinner and undivided. The first sign of the limb-bud itself appears at stage 38. 2. A whole, a half or a third of a limb rudiment can develop into a normal or reduplicated limb when transplanted to the flank. 3. An anterior half of a limb rudiment fails to develop when transplanted to the head but will do so if accompanied by a small piece of flank tissue. 4. Small pieces of tissue from a wide area of the flank will cause reduplication of the forelimb if grafted to the anterior margin of the rudiment. It is shown that the whole of the reduplication is formed from host tissue and has the morphological character of the host. 5. Reduplications have posterior structures arranged symmetrically on both sides of the midline. Both muscles and cartilages are duplicated. 6. It is suggested that the same interaction between prospective flank and limb is responsible for the capacity for growth on the head, the induction of reduplications and the formation of the anteroposterior pattern of the limb in normal development. 7. A simple rule is proposed which explains the occurrence of reduplications in classical work on the amphibian limb.

Development ◽  
1996 ◽  
Vol 122 (8) ◽  
pp. 2319-2330 ◽  
Author(s):  
M.A. Ros ◽  
A. Lopez-Martinez ◽  
B.K. Simandl ◽  
C. Rodriguez ◽  
J.C. Izpisua Belmonte ◽  
...  

We have analyzed the pattern of expression of several genes implicated in limb initiation and outgrowth using limbless chicken embryos. We demonstrate that the expressions of the apical ridge associated genes, Fgf-8, Fgf-4, Bmp-2 and Bmp-4, are undetectable in limbless limb bud ectoderm; however, FGF2 protein is present in the limb bud ectoderm. Shh expression is undetectable in limbless limb bud mesoderm. Nevertheless, limbless limb bud mesoderm shows polarization manifested by the asymmetric expression of Hoxd-11, −12 and −13, Wnt-5a and Bmp-4 genes. The posterior limbless limb bud mesoderm, although not actually expressing Shh, is competent to express it if supplied with exogenous FGF or transplanted to a normal apical ridge environment, providing further evidence of mesodermal asymmetry. Exogenous FGF applied to limbless limb buds permits further growth and determination of recognizable skeletal elements, without the development of an apical ridge. However, the cells competent to express Shh do so at reduced levels; nevertheless, Bmp-2 is then rapidly expressed in the posterior limbless mesoderm. limbless limb buds appear as bi-dorsal structures, as the entire limb bud ectoderm expresses Wnt-7a, a marker for dorsal limb bud ectoderm; the ectoderm fails to express En-1, a marker of ventral ectoderm. As expected, C-Lmx1, which is downstream of Wnt-7a, is expressed in the entire limbless limb bud mesoderm. We conclude that anteroposterior polarity is established in the initial limb bud prior to Shh expression, apical ridge gene expression or dorsal-ventral asymmetry. We propose that the initial pattern of gene expressions in the emergent limb bud is established by axial influences on the limb field. These permit the bud to emerge with asymmetric gene expression before Shh and the apical ridge appear. We report that expression of Fgf-8 by the limb ectoderm is not required for the initiation of the limb bud. The gene expressions in the pre-ridge limb bud mesoderm, as in the limb bud itself, are unstable without stimulation from the apical ridge and the polarizing region (Shh) after budding is initiated. We propose that the defect in limbless limb buds is the lack of a dorsal-ventral interface in the limb bud ectoderm where the apical ridge induction signal would be received and an apical ridge formed. These observations provide evidence for the hypothesis that the dorsal-ventral ectoderm interface is a precondition for apical ridge formation.


Development ◽  
1999 ◽  
Vol 126 (21) ◽  
pp. 4817-4826 ◽  
Author(s):  
C.J. Neumann ◽  
H. Grandel ◽  
W. Gaffield ◽  
S. Schulte-Merker ◽  
C. Nusslein-Volhard

Sonic hedgehog (Shh) is expressed in the posterior vertebrate limb bud mesenchyme and directs anteroposterior patterning and growth during limb development. Here we report an analysis of the pectoral fin phenotype of zebrafish sonic you mutants, which disrupt the shh gene. We show that Shh is required for the establishment of some aspects of anteroposterior polarity, while other aspects of anteroposterior polarity are established independently of Shh, and only later come to depend on Shh for their maintenance. We also demonstrate that Shh is required for the activation of posterior HoxD genes by retinoic acid. Finally, we show that Shh is required for normal development of the apical ectodermal fold, for growth of the fin bud, and for formation of the fin endoskeleton.


Amongst the Fellows elected to the Royal Society in 1941 were W. T. Astbury for his studies using X-ray analysis to study the structures of natural fibres, and amongst the Foreign Members elected that year was Ross G. Harrison for his contributions to embryology. Astbury and Harrison were very different in temperament, and worked in very different fields on either side of the Atlantic, yet they were united in their approach to the study of biological phenomena. Both Astbury and Harrison believed that the organization and form of biological materials whether wool fibres or the limb-bud in an amphibian embryo depended on molecular structure and pattern. Moreover both were concerned with dynamic aspects of form; Astbury’s greatest achievement was to demonstrate the dynamic, reversible folding and stretching of proteins in the k-m-e-f group, and Harrison looked to changing molecular patterns to account for changing symmetries in the developing embryo. It was this common approach that brought them together and led to Harrison spending a brief month in Leeds where they and K. M. Rudall performed what have been described as ‘truly progressive experiments in molecular biology’. I believe this short series of experiments illuminates the character and work of both Harrison and Astbury and illustrates the difficulties, practical and conceptual, in carrying out ‘progressive experiments’. I shall begin by reviewing briefly the embryological background of the time before going on to discuss in detail the approaches of Harrison and Astbury to their work and the outcome of their collaboration.


Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 199-206 ◽  
Author(s):  
A. Vogel ◽  
C. Tickle

The polarizing region is a major signalling tissue involved in patterning the tissues of the vertebrate limb. The polarizing region is located at the posterior margin of the limb bud and can be recognized by its ability to induce additional digits when grafted to the anterior margin of a chick limb bud. The signal from the polarizing region operates at the tip of the bud in the progress zone, a zone of undifferentiated mesenchymal cells, maintained by interactions with the apical ectodermal ridge. A number of observations have pointed to a link between the apical ectodermal ridge and signalling by the polarizing region. To test this possibility, we removed the posterior apical ectodermal ridge of chick wing buds and assayed posterior mesenchyme for polarizing activity. When the apical ectodermal ridge is removed, there is a marked decrease in polarizing activity of posterior cells. The posterior apical ectodermal ridge is known to express FGF-4 and we show that the decrease in polarizing activity of posterior cells of wing buds that normally follows ridge removal can be prevented by implanting a FGF-4-soaked bead. Furthermore, we show that both ectoderm and FGF-4 maintain polarizing activity of limb bud cells in culture.


Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 105-125
Author(s):  
Madeleine Gumpel-Pinot ◽  
D. A. Ede ◽  
O. P. Flint

Fragments of quail wing bud containing myogenic cells of somitic origin and fragments of quail sphlanchopleural tissue were introduced into the interior of the wing bud of fowl embryo hosts. No movement of graft into host tissue occurred in the control, but myogenic cells from the quail wing bud fragments underwent long migrations in an apical direction to become incorporated in the developing musculature of the host. When the apical ectodermal ridge (AER), together with some subridge mesenchyme, was removed at the time of grafting, no such cell migration occurred. The capacity of grafted myogenic cells to migrate in the presence of AER persists to H.H. stage 25, when myogenesis has begun, but premyogenic cells in the somites, which normally migrate out into the early limb bud, do not migrate when somite fragments are grafted into the wing bud. Coelomic grafts of apical and proximal wing fragments showed that apical sections of quail wing buds become invaded by myogenic cells of the host, but grafts from proximal wing bud regions do not.


2020 ◽  
Vol 1 ◽  
pp. 32-36
Author(s):  
I. V. Botantsov ◽  

According to the Constitution of the Russian Federation, every citizen has the right to freedom of movement on its territory, but due to the fact that minors cannot be held accountable for their actions, there is a need to control their movement by legal representatives. The practical determination of the age of independent travel of minors and the issues of drawing up documents by parents authorizing them to do so are the subjects of disputes that are subject to judicial resolution. The article provides an analysis of the relevant practice, accompanied by the author's comments.


Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 257-265
Author(s):  
Amata Hornbruch ◽  
Lewis Wolpert

Hensen's node from stage 4 to stage 10 shows polarizing activity when grafted to the anterior margin of the chick limb bud. It can specify additional digits though its action is somewhat attenuated when compared with the effect of a grafted polarizing region. At stage 10 the activity disappears from the node and is found both posterior to the node and in the future wing region of the flank. The ability of Hensen's node to generate a positional signal suggests that the signal in the limb and early embryo may be similar. The results support the view of the polarizing region as a discrete signalling region.


2016 ◽  
Vol 47 (2) ◽  
pp. 191
Author(s):  
Andrew Robertson

In the law of contract questions of risk allocation properly turn, where possible, on interpretation of the agreement. This article will explore the limits of that approach. It will do so by considering two doctrines that lie at the boundaries of contract interpretation: the implication of terms in fact and the remoteness principle. Both doctrines have been commonly understood as gap-filling rules, but in two influential judgments Lord Hoffmann sought to recast them as interpretative principles. It will be argued in this article that the implication of terms in fact can properly be regarded as an interpretative exercise, but the same cannot be said of the application of the remoteness doctrine. The implication of terms in fact can helpfully be understood as interpretative, provided care is taken to explain the paths of reasoning leading to the conclusion that a contractual instrument must be understood to include a particular unexpressed term. Because no common paths of interpretative reasoning can be identified for the determination of remoteness questions, it is unsettling and counterproductive to attempt to take an interpretative approach to these questions.


Development ◽  
1974 ◽  
Vol 32 (3) ◽  
pp. 819-833
Author(s):  
M. C. Prestige ◽  
Margaret A. Wilson

1. The development of the ventral root (VR) in Xenopus has been studied by electron microscopy. Total fibre counts, and counts of classes of fibres were made from large photomontages of the whole of VR 9 at × 15000. 2. The total number of fibres in the root shows the same pattern of initial rise, peak, and subsequent decline that previous ventral horn (VH) cell counts had shown, The two curves overlay each other initially, but after the decline, there were apparently more cells than fibres. 3. Promyelin and myelin formation was seen at the time of the decline. There was no evidence that dying axons had started to myelinate. 4. In some animals the limb-bud was removed at the time of its first penetration by nerve fibres. The ventral roots developed normally for a week, but thereafter fibre loss was accentuated, advanced and more profound, so that after another week, no fibres were left. In these roots, no promyelin or myelin was formed. 5. In other animals, it was shown that there is no evidence for collateral sprouting in the ventral roots during normal development. 6. It is argued that the axons which die in normal development have already reached the limb-bud. 7. The correspondence between axon and cell number is discussed.


Development ◽  
1994 ◽  
Vol 120 (9) ◽  
pp. 2619-2628 ◽  
Author(s):  
R. Kuraishi ◽  
L. Osanai

Contribution of maternal cytoplasmic factors and cellular interaction to determination of archenteron in a starfish embryo was analyzed by (1) examining temporal and positional pattern of expression of an endoderm-specific enzyme, alkaline phosphatase, (2) deleting the vegetal polar fragment from an immature oocyte and (3) changing the orientation of a blastomere within an early stage embryo. The archenteron (and the differentiated digestive tract) of Asterina pectinifera was divided into three areas based on the time of start of alkaline phosphatase expression. At 27 hours after 1-methyladenine treatment, the whole archenteron except the anterior end started to express alkaline phosphatase. The anterior negative area differentiated into mesodermal tissues such as mesenchyme cells and anterior coelomic pouches (anterior mesodermal area). The alkaline-phosphatase-positive area 1 gave rise to the esophagus and the anterior end of the stomach. Alkaline-phosphatase-positive area 2, which was gradually added to the posterior end of the archenteron after 30 hours, became alkaline-phosphatase- positive and formed the middle-to-posterior part of the stomach and the intestine. When the vegetal oocyte fragment, the volume of which was more than 8% of that of the whole oocyte, was removed from the immature oocyte, archenteron formation was strongly suppressed. However, when the volume deleted was less than 6%, most of the larvae started archenteron formation before the intact controls reached the mesenchyme-migration stage (30 hours). Although cells in the alkaline-phosphatase-positive area 2 are added to the posterior end of the archenteron after 30 hours in normal development (R. Kuraishi and K. Osanai (1992) Biol. Bull. Mar. Biol. Lab., Woods Hole 183, 258–268), few larvae started gastrulation after 30 hours. Estimation of the movement of the oocyte cortex during the early development suggested that the area that inherits the cortex of the 7% area coincides with the combined area of anterior mesodermal area and alkaline-phosphatase-positive area 1. When one of the blastomeres was rotated 180° around the axis of apicobasal polarity at the 2-cell stage to make its vegetal pole face the animal pole of the other blastomere, two archentera formed at the separated vegetal poles. Intracellular injection of tracers showed that cells derived from the animal blastomere, which gives rise to the ectoderm in normal development, stayed in the outer layer until 30 hours; a proportion of them then entered the archenteron gradually. The involuted animal cells expressed alkaline phosphatase and were incorporated into the middle-to-posterior part of the stomach and the intestine. These results suggest that anterior mesodermal area and alkaline-phosphatase-positive area 1 are determined by cytoplasmic factor(s) that had already been localized in their presumptive areas. In contrast, alkaline-phosphatase-positive area 2 becomes the endoderm by homoiogenetic induction from the neighboring area on the vegetal side, namely alkaline-phosphatase-positive area 1.


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