scholarly journals The effect of local application of retinoic acid to the anterior margin of the developing chick limb

Development ◽  
1983 ◽  
Vol 78 (1) ◽  
pp. 269-289
Author(s):  
Dennis Summerbell
Keyword(s):  
Retinoic Acid ◽  
Limb Development ◽  
Anterior Margin ◽  
Mirror Image ◽  
Limb Bud ◽  
Local Application ◽  
Low Doses ◽  
Full Dose ◽  
Early Stages ◽  
High Doses

Local application of retinoic acid to the chick limb bud produces effects that are dose and/or stage dependent. Low doses and/or old stages tend to give normal limbs or perhaps one or two supernumerary digits of a more anterior character. Medium doses and/or intermediate stages tend to give full mirror-image supernumeraries with two or even three extra digits including particularly digits of a posterior character. High doses and/or early stages give limbs in which supernumerary digits fail to form or are lost, and in which even host skeletal elements are missing or reduced. The effects are graded over the full dose and/or stage range. Various explanations are discussed in the context of the current hypotheses of limb development. We conclude that one should not necessarily interpret the results as evidence that retinoids normally play a part in the control of development or regeneration.

The chick limbless mutation causes abnormalities in limb bud dorsal-ventral patterning: implications for the mechanism of apical ridge formation

Development ◽  
1996 ◽  
Vol 122 (12) ◽  
pp. 3851-3861 ◽  
Author(s):  
U. Grieshammer ◽  
G. Minowada ◽  
J.M. Pisenti ◽  
U.K. Abbott ◽  
G.R. Martin
Keyword(s):  
Gene Expression ◽  
Limb Development ◽  
Positional Information ◽  
Limb Bud ◽  
Local Application ◽  
Chick Embryos ◽  
Limb Buds ◽  
Early Stages ◽  
Potential Link

In chick embryos homozygous for the limbless mutation, limb bud outgrowth is initiated, but a morphologically distinct apical ridge does not develop and limbs do not form. Here we report the results of an analysis of gene expression in limbless mutant limb buds. Fgf4, Fgf8, Bmp2 and Msx2, genes that are expressed in the apical ridge of normal limb buds, are not expressed in the mutant limb bud ectoderm, providing molecular support for the hypothesis that limb development fails in the limbless embryo because of the inability of the ectoderm to form a functional ridge. Moreover, Fgf8 expression is not detected in the ectoderm of the prospective limb territory or the early limb bud of limbless embryos. Since the early stages of limb bud outgrowth occur normally in the mutant embryos, this indicates that FGF8 is not required to promote initial limb bud outgrowth. In the absence of FGF8, Shh is also not expressed in the mutant limb buds, although its expression can be induced by application of FGF8-soaked beads. These observations support the hypothesis that Fgf8 is required for the induction of Shh expression during normal limb development. Bmp2 expression was also not detected in mutant limb mesoderm, consistent with the hypothesis that SHH induces its expression. In contrast, SHH is not required for the induction of Hoxd11 or Hoxd13 expression, since expression of both these genes was detected in the mutant limb buds. Thus, some aspects of mesoderm A-P patterning can occur in the absence of SHH and factors normally expressed in the apical ridge. Intriguingly, mutant limbs rescued by local application of FGF displayed a dorsalized feather pattern. Furthermore, the expression of Wnt7a, Lmx1 and En1, genes involved in limb D-V patterning, was found to be abnormal in mutant limb buds. These data suggest that D-V patterning and apical ridge formation are linked, since they show that the limbless mutation affects both processes. We present a model that explains the potential link between D-V positional information and apical ridge formation, and discuss the possible function of the limbless gene in terms of this model.

scholarly journals Retinoids reprogramme pre-bud mesenchyme to give changes in limb pattern

Development ◽  
1987 ◽  
Vol 100 (4) ◽  
pp. 723-733 ◽  
Author(s):  
S.M. Wilde ◽  
S.E. Wedden ◽  
C. Tickle
Keyword(s):  
Retinoic Acid ◽  
Effective Treatment ◽  
Limb Development ◽  
Apical Ectodermal Ridge ◽  
Limb Bud ◽  
Chick Embryos ◽  
Early Stages ◽  
Limb Pattern

Retinoic acid was locally applied to presumptive limb regions of chick embryos to find out the earliest time at which the limb pattern can be reprogrammed. When beads soaked in retinoic acid were placed in the appropriate positions in embryos at stage 10 or older, duplicated or reduced leg patterns resulted. To pin point the time at which the cells in the limb rudiment respond to the retinoid, beads were removed at various times and the lengths of exposure required to reprogramme limb development found. The early limb rudiments require longer exposures to give duplications than late rudiments. The effective treatment periods last at least until stage 17 when the limb bud and apical ectodermal ridge develop. In contrast, the length of exposure to reduce the limb is constant at early stages. Retinoids first start acting to produce duplicated structures between stages 10 and 13. Therefore, retinoids appear to begin to reprogramme the cells as soon as they are determined to give rise to a limb.

Retinoic acid and pattern formation in the developing chick wing: SEM and quantitative studies of early effects on the apical ectodermal ridge and bud outgrowth

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 139-169
Author(s):  
J. Lee ◽  
C. Tickle
Keyword(s):  
Retinoic Acid ◽  
Local Application ◽  
Local Source ◽  
Local Concentration ◽  
Low Doses ◽  
Response Curves ◽  
Wing Patterns ◽  
Skeletal Pattern ◽  
High Doses ◽  
Dose Dependent

When retinoic acid is locally applied to the anterior margin of developing chick wing buds on ion-exchange beads, dose-dependent changes in the skeletal pattern result. At low doses, additional digits develop. At high doses, there is thinning of the symmetrical wing. Local application of retinoic acid to the apex of the bud also leads to pattern changes, but in contrast normal wing patterns are almost always obtained following application posteriorly. These effects are manifest at 6–7 days after the operation although only a brief exposure (14–20 h) to retinoic acid is required. Therefore the morphology of wing buds was studied at shorter times after the start of treatment. The local application of retinoic acid to the wing bud margin leads to changes in extent of the apical ridge that can be detected at 24 h after application. The behaviour of the apical ridge with varying doses and positions of retinoic acid application has been analysed quantitatively and dose response curves obtained. At low doses of retinoic acid, the length of the apical ridge increases or remains constant, but then progressively decreases with higher doses. The progressive obliteration of the ridge starts first near the bead and then involves more distant parts of the bud. Thus the region of the ridge affected depends on the position at which the retinoic acid is applied. We propose that these effects on the apical ridge reflect dose-dependent responses to the local concentration of retinoic acid that varies with distance from the source. At high doses, the apical ridge disappears but at low doses it is maintained. Since grafts of polarizing region tissue also have a graded effect on ridge morphology, a possible interpretation of the retinoic acid effects is that tissue adjacent to the source is converted into polarizing region tissue. Alternatively, retinoic acid may act directly on the ridge cells. The changes in the extent of the apical ridge produced by retinoic acid lead to different forms of bud outgrowth. The form of the outgrowth depends on the dose of retinoic acid, the position of application and the interaction between the effects of the local source of retinoic acid and those of the polarizing region of the host bud. These considerations give some insights into why anterior application of retinoic acid leads to the development of additional digits whereas posterior application generally gives normal wings.

Retinoic acid signaling is required during early chick limb development

Development ◽  
1996 ◽  
Vol 122 (5) ◽  
pp. 1385-1394 ◽  
Author(s):  
J.A. Helms ◽  
C.H. Kim ◽  
G. Eichele ◽  
C. Thaller
Keyword(s):  
Retinoic Acid ◽  
Limb Development ◽  
Acid Synthesis ◽  
Apical Ectodermal Ridge ◽  
Limb Bud ◽  
Retinoid Receptor ◽  
Limb Morphogenesis ◽  
Wing Bud ◽  
Zone Of Polarizing Activity

In the chick limb bud, the zone of polarizing activity controls limb patterning along the anteroposterior and proximodistal axes. Since retinoic acid can induce ectopic polarizing activity, we examined whether this molecule plays a role in the establishment of the endogenous zone of polarizing activity. Grafts of wing bud mesenchyme treated with physiologic doses of retinoic acid had weak polarizing activity but inclusion of a retinoic acid-exposed apical ectodermal ridge or of prospective wing bud ectoderm evoked strong polarizing activity. Likewise, polarizing activity of prospective wing mesenchyme was markedly enhanced by co-grafting either a retinoic acid-exposed apical ectodermal ridge or ectoderm from the wing region. This equivalence of ectoderm-mesenchyme interactions required for the establishment of polarizing activity in retinoic acid-treated wing buds and in prospective wing tissue, suggests a role of retinoic acid in the establishment of the zone of polarizing activity. We found that prospective wing bud tissue is a high-point of retinoic acid synthesis. Furthermore, retinoid receptor-specific antagonists blocked limb morphogenesis and down-regulated a polarizing signal, sonic hedgehog. Limb agenesis was reversed when antagonist-exposed wing buds were treated with retinoic acid. Our results demonstrate a role of retinoic acid in the establishment of the endogenous zone of polarizing activity.

Etudes expérimentales sur le rôle des somites au cours des premiers stades du développement du membre antérieur chez l'embryon du Chélonien Emys orbicularis L.

Development ◽  
1974 ◽  
Vol 32 (2) ◽  
pp. 417-430
Author(s):  
Par J. Vasse
Keyword(s):  
Experimental Study ◽  
Developmental Stage ◽  
Limb Development ◽  
Limb Bud ◽  
Stages Of Development ◽  
Emys Orbicularis ◽  
Bud Development ◽  
Early Stages

Experimental study on the role of the somites during the early stages of development of the front limbs of the embryo of the chelonian Emys orbicularis L. Ablation of postotic somites 6–13 on one side in embryos of Emys orbicularis L. at the developmental stage when 20–23 somite pairs were present, led to arrest of forelimb-bud development in the somatopleure adjacent to the ablated somites on the operated side. Limb development in the somatopleure adjacent to intact somites on the operated side was unaffected, attaining the same stage as on the non-operated side. Ablation at later stages (25–33 somite pairs) did not prevent development of the limb adjacent to the ablated somites. When a part of the prospective somatopleure was injured, the remaining part formed a small limb-bud. When an obstacle was placed between the somatopleural mesoderm and the adjacent somite, development of the somatopleure stopped at this level. These results corroborate those obtained from previous studies in various reptilian embryos concerning the role of the ventral somite extensions as activators of proliferation in the somatopleural mesoderm. Injury to the ventral extension alone led to serious disturbances in the somatopleural mesoderm adjacent to this somite.

Retinoids and pattern formation in a hydroid

Development ◽  
1984 ◽  
Vol 81 (1) ◽  
pp. 253-271
Author(s):  
W. A. Müller
Keyword(s):  
Retinoic Acid ◽  
Prolonged Treatment ◽  
Low Doses ◽  
Response Curves ◽  
Head Formation ◽  
Pulse Type ◽  
Head Activator ◽  
High Doses ◽  
Pattern Specification ◽  
Hydroid Polyps

The retinoids (retinol, retinal, retinoic acid) cause alterations in the pattern of limb elements in vertebrates (Summerbell & Harvey, 1983). As shown here, retinoids also influence pattern specification in hydroid polyps (Hydractinia echinata) in a way suggesting interference with the generation and transmission of signals responsible for the dimension and spacing of structures. A pulse-type application of low doses (e.g. retinoic acid 10-6 to 10-10 M, 4 h) causes metamorphosing primary polyps to develop more tentacles but fewer stolons per unit circumference, to shorten the length of the hydranth while the stolon elongates, and to bud secondary hydranths at high frequency 2–3 days after treatment (Fig. 3). Dose-response curves display optimum peaks. It is argued that the increase in budding rate is due to a reduction of the range of spacing signals emitted by the primary hydranth. In regenerating hydranths, low doses (10−10 to 10−9M) improve the rate of head formation, whilst medium doses (10−8 to 10−6M) result in more tentacles being regenerated. However, prolonged treatment with high doses (10−6 to 10−5 M) causes the animals to reduce all head structures and to transform eventually into stolons, in contravention of the rule of distal transformation that they normally obey (Fig. 8). The effects of the retinoids are counteracted by a putative morphogen, the endogenous inhibitor isolated from Hydra by Berking (1977). The Hydra-derived ‘head-activator’ displayed no stimulating effect on the number of tentacles and buds formed.

Retinoic acid induces a pattern of digits in anterior half wing buds that lack the zone of polarizing activity

Development ◽  
1989 ◽  
Vol 107 (4) ◽  
pp. 863-867 ◽  
Author(s):  
G. Eichele
Keyword(s):  
Retinoic Acid ◽  
Cell Fate ◽  
Anterior Margin ◽  
Limb Bud ◽  
Posterior Half ◽  
Anterior Half ◽  
Zone Of Polarizing Activity

Wing buds whose posterior half is excised, develop into wings lacking distal structures. However, such experimentally generated preaxial half wing buds can be rescued by implanting a retinoic-acid-releasing bead at their anterior margin. The polarity of the pattern that originates from preaxial half wing buds is reversed. For example, instead of a 234 digit pattern typical for normal wings, the order of digits is 432. This result implies that retinoic acid has the capacity to reprogram anterior limb bud tissue, and that the resulting change in cell fate does not depend on the presence of posterior tissue regions such as the zone of polarizing activity (ZPA).

Bone morphogenetic proteins and a signalling pathway that controls patterning in the developing chick limb

Development ◽  
1994 ◽  
Vol 120 (1) ◽  
pp. 209-218 ◽  
Author(s):  
P.H. Francis ◽  
M.K. Richardson ◽  
P.M. Brickell ◽  
C. Tickle
Keyword(s):  
Gene Expression ◽  
Retinoic Acid ◽  
Bone Morphogenetic Proteins ◽  
Limb Development ◽  
Posterior Part ◽  
Signalling Pathway ◽  
Limb Bud ◽  
Bone Morphogenetic Protein 2 ◽  
Close Relationship ◽  
Wing Bud

We show here that bone morphogenetic protein 2 (BMP-2) is involved in patterning the developing chick limb. During early stages of limb development, mesenchymal expression of the Bmp-2 gene is restricted to the posterior part of the bud, in a domain that colocalizes with the polarizing region. The polarizing region is a group of cells at the posterior margin of the limb bud that can respecify the anteroposterior axis of the limb when grafted anteriorly and can activate expression of genes of the HoxD complex. We dissect possible roles of BMP-2 in the polarizing region signalling pathway by manipulating the developing wing bud. Retinoic acid application, which mimics the effects of polarizing region grafts, activates Bmp-2 gene expression in anterior cells. This shows that changes in anteroposterior pattern are correlated with changes in Bmp-2 expression. When polarizing region grafts are placed at the anterior margin of the wing bud, the grafts continue to express the Bmp-2 gene and also activate Bmp-2 expression in the adjacent anterior host mesenchyme. These data suggest that BMP-2 is part of the response pathway to the polarizing signal, rather than being the signal itself. In support of this, BMP-2 protein does not appear to have any detectable polarizing activity when applied to the wing bud. The pattern of Bmp-4 gene expression in the developing wing bud raises the possibility that BMP-2 and BMP-4 could act in concert. There is a close relationship, both temporal and spatial, between the activation of the Bmp-2 and Hoxd-13 genes in response to retinoic acid and polarizing region grafts, suggesting that expression of the two genes might be linked.

Role of dHAND in the anterior-posterior polarization of the limb bud: implications for the Sonic hedgehog pathway

Development ◽  
2000 ◽  
Vol 127 (10) ◽  
pp. 2133-2142 ◽  
Author(s):  
M. Fernandez-Teran ◽  
M.E. Piedra ◽  
I.S. Kathiriya ◽  
D. Srivastava ◽  
J.C. Rodriguez-Rey ◽  
...  
Keyword(s):  
Sonic Hedgehog ◽  
Limb Development ◽  
Mirror Image ◽  
Limb Bud ◽  
Bhlh Transcription Factor ◽  
Cardiovascular Development ◽  
Anterior Border ◽  
Lateral Plate ◽  
Shh Pathway ◽  
Anterior Posterior

dHAND is a basic helix-loop-helix (bHLH) transcription factor essential for cardiovascular development. Here we analyze its pattern of expression and functional role during chick limb development. dHAND expression was observed in the lateral plate mesoderm prior to emergence of the limb buds. Coincident with limb initiation, expression of dHAND became restricted to the posterior half of the limb bud. Experimental procedures that caused mirror-image duplications of the limb resulted in mirror-image duplications of the pattern of dHAND expression along the anterior-posterior axis. Retroviral overexpression of dHAND in the limb bud produced preaxial polydactyly, corresponding to mild polarizing activity at the anterior border. At the molecular level, misexpression of dHAND caused ectopic activation of members of the Sonic hedgehog (Shh) pathway, including Gli and Patched, in the anterior limb bud. A subset of infected embryos displayed ectopic anterior activation of Shh. Other factors implicated in anterior-posterior polarization of the bud such as the most 5′ Hoxd genes and Bmp2 were also ectopically activated at the anterior border. Our results indicate a role for dHAND in the establishment of anterior-posterior polarization of the limb bud.

Transient establishment of anteroposterior polarity in the zebrafish pectoral fin bud in the absence of sonic hedgehog activity

Development ◽  
1999 ◽  
Vol 126 (21) ◽  
pp. 4817-4826 ◽  
Author(s):  
C.J. Neumann ◽  
H. Grandel ◽  
W. Gaffield ◽  
S. Schulte-Merker ◽  
C. Nusslein-Volhard
Keyword(s):  
Retinoic Acid ◽  
Sonic Hedgehog ◽  
Limb Development ◽  
Normal Development ◽  
Limb Bud ◽  
Pectoral Fin ◽  
Anteroposterior Patterning ◽  
Vertebrate Limb ◽  
Anteroposterior Polarity

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.

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