Distribution of polarizing activity and potential for limb formation in mouse and chick embryos and possible relationships to polydactyly

M. Tanaka; M.J. Cohn; P. Ashby; M. Davey; P. Martin; C. Tickle

doi:10.1242/dev.127.18.4011

Distribution of polarizing activity and potential for limb formation in mouse and chick embryos and possible relationships to polydactyly

Development ◽

10.1242/dev.127.18.4011 ◽

2000 ◽

Vol 127 (18) ◽

pp. 4011-4021 ◽

Cited By ~ 1

Author(s):

M. Tanaka ◽

M.J. Cohn ◽

P. Ashby ◽

M. Davey ◽

P. Martin ◽

...

Keyword(s):

Body Plan ◽

Mouse Embryos ◽

Apical Ectodermal Ridge ◽

Central Feature ◽

Chick Embryos ◽

Limb Buds ◽

Naturally Occurring ◽

Induction Signal

A central feature of the tetrapod body plan is that two pairs of limbs develop at specific positions along the head-to-tail axis. However, the potential to form limbs in chick embryos is more widespread. This could have implications for understanding the basis of limb abnormalities. Here we extend the analysis to mouse embryos and examine systematically the potential of tissues in different regions outside the limbs to contribute to limb structures. We show that the ability of ectoderm to form an apical ridge in response to FGF4 in both mouse and chick embryos exists throughout the flank as does ability of mesenchyme to provide a polarizing region signal. In addition, neck tissue has weak polarizing activity. We show, in chick embryos, that polarizing activity of tissues correlates with the ability either to express Shh or to induce Shh expression. We also show that cells from chick tail can give rise to limb structures. Taken together these observations suggest that naturally occurring polydactyly could involve recruitment of cells from regions adjacent to the limb buds. We show that cells from neck, flank and tail can migrate into limb buds in response to FGF4, which mimics extension of the apical ectodermal ridge. Furthermore, when we apply simultaneously a polarizing signal and a limb induction signal to early chick flank, this leads to limb duplications.

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Ultrastructural aspects of early development of the fore-limb buds in the chick and the mouse

Proceedings of the Royal Society of London Series B Biological Sciences ◽

10.1098/rspb.1965.0045 ◽

1965 ◽

Vol 162 (988) ◽

pp. 387-405 ◽

Cited By ~ 50

Keyword(s):

Endoplasmic Reticulum ◽

Early Development ◽

Secretory Granules ◽

Free Cell ◽

Mouse Embryos ◽

Limb Bud ◽

Chick Embryos ◽

Limb Buds ◽

Mouse Limb ◽

A Chain

Light microscope investigations of the early development of the fore-limb buds in chick and mouse were made to guide electron microscope studies with these tissues. At the time of maximal development of the ectodermal apical ridge there is a higher concentration of cytoplasmic RNA in the apical ridge cells than in the other cells of the limb bud. Ultrastructural investigations showed that, in the mesoblast cells at the earliest stages, profiles of endoplasmic reticulum are often found attached to the outer nuclear membrane. Some what later, discontinuities of nuclear envelope occur by which the content of the nucleus may communicate with the endoplasmic reticulum. In the cytoplasm of the mesoblast cells at these stages there were many granules similar in form and size to secretory granules of gland cells. Ribosomes are in the polysomal condition. At stages later than 20 in chick and in 11-day-old mouse embryos, the mesoblast shows the character of a syncytial tissue. Epiblast cells possess all the characters of an epithelium with well developed junctional complexes. The desmosomes form a chain consisting of units equipped with individual dense plaques, but connected by continuous bundles of fibres running parallel to the chain. The free cell membrane of the epiblastic cells, particularly at early stages, forms numerous microvilli and single cilia. In later stages during the form action of the ectodermal apical ridge, cilia have been found between the cells. This fact indicates that when the apical ridge is formed ectodermal cells migrate towards the margin of the limb bud. At these stages microvilli are also found between the apical ridge cells where they contribute to the cell-to-cell adhesion. Beginning at stage 22 in chick embryos and from the 12th day in mouse embryos there are in cells of the apical ridge numerous and extensive Golgi systems spread throughout the cytoplasm. Some what later there appear successively lysosomes, cytolysomes and extranuclear necrotic centres. All these organelles manifest acid phosphatase activity and are thoughtto initiate the involutive process in the apical ridge. Pycnosis and karyorrhexis appear as the last stage of cellular degeneration. Degenerating cells undergo phagocytosis by neighbouring epithelial cells. A basement membrane is present at all stages of development of the chick and mouse limb buds. It is an acellular continuous structure lining the internal (basal) surface of the epiblast, but in chick embryos it shows discontinuities immediately under the apical ectodermal ridge at the time of its maximum development.

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Altered expression of the chicken homeobox-containing genes GHox-7 and GHox-8 in the limb buds of limbless mutant chick embryos

Development ◽

10.1242/dev.113.4.1487 ◽

1991 ◽

Vol 113 (4) ◽

pp. 1487-1493 ◽

Cited By ~ 13

Author(s):

C.N. Coelho ◽

K.M. Krabbenhoft ◽

W.B. Upholt ◽

J.F. Fallon ◽

R.A. Kosher

Keyword(s):

Proper Time ◽

Blot Hybridization ◽

Development Stage ◽

Apical Ectodermal Ridge ◽

Limb Bud ◽

Chick Embryos ◽

Dot Blot ◽

Limb Buds ◽

Altered Expression

It has been suggested that the reciprocal expression of the chicken homeobox-containing genes GHox-8 and GHox-7 by the apical ectodermal ridge and subjacent limb mesoderm might be involved in regulating the proximodistal outgrowth of the developing chick limb bud. In the present study the expression of GHox-7 and GHox-8 has been examined by in situ and dot blot hybridization in the developing limb buds of limbless mutant chick embryos. The limb buds of homozygous mutant limbless embryos form at the proper time in development (stage 17/18), but never develop an apical ectodermal ridge, fail to undergo normal elongation, and eventually degenerate. At stage 18, which is shortly following the formation of the limb bud, the expression of GHox-7 is considerably reduced (about 3-fold lower) in the mesoderm of limbless mutant limb buds compared to normal limb bud mesoderm. By stages 20 and 21, as the limb buds of limbless embryos cease outgrowth, GHox-7 expression in limbless mesoderm declines to very low levels, whereas GHox-7 expression increases in the mesoderm of normal limb buds which are undergoing outgrowth. In contrast to GHox-7, expression of GHox-8 in limbless mesoderm at stage 18 is quantitatively similar to its expression in normal limb bud mesoderm, and in limbless and normal mesoderm GHox-8 expression is highly localized in the anterior mesoderm of the limb bud. In normal limb buds, GHox-8 is also expressed in high amounts by the apical ectodermal ridge.(ABSTRACT TRUNCATED AT 250 WORDS)

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Early changes in limb buds of chick embryos after thalidomide treatment

Development ◽

10.1242/dev.16.2.289 ◽

1966 ◽

Vol 16 (2) ◽

pp. 289-300

Author(s):

A. Jurand

Keyword(s):

Embryonic Development ◽

Experimental Investigations ◽

Experimental Production ◽

Chick Embryos ◽

Teratogenic Effects ◽

Limb Buds ◽

Experimental Animals ◽

Almost All ◽

Thalidomide Treatment

Since the first observations of hypoplastic and aplastic thalidomide deformities in infants (McBride, 1961; Lenz, 1962), the literature on this subject has grown to many hundreds of communications. Experimental investigations in almost all cases have been undertaken to show whether thalidomide and its metabolites have any teratogenic effects in experimental animals. Numerous review papers are available on this subject, e.g. Giroud, Tuchmann-Duplessis & Mercier-Parot (1962), Somers (1963), and Salzgeber & Wolff (1964). Chick embryos did not seem for some time to be suitable for experimental production of typical thalidomide deformities. However, Kemper (1962a, b), Yang, Yang & Liang (1962). Boylen, Home & Johnson (1963) and Leone (1963) have shown that thalidomide can produce a whole range of ectromelian deformities provided that it is introduced into the egg at a particular period of embryonic development.

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Effects of some Amino-acid and Purine Antagonists on Chick Embryos

Development ◽

10.1242/dev.6.2.365 ◽

1958 ◽

Vol 6 (2) ◽

pp. 365-372

Author(s):

C. H. Waddington ◽

Margaret Perry

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Effective Concentration ◽

Metabolic Inhibitors ◽

Chick Embryos ◽

Purine Analogue ◽

Know How ◽

Naturally Occurring ◽

Different Types ◽

Strong Action

Several authors have studied the effects on developing embryos of substances which are analogues of naturally occurring amino-acids and purines, and known to act, in other systems, as metabolic inhibitors. It was emphasized by Waddington, Feldman, & Perry (1955) that any particular substance may exhibit very different effects in embryos of different types. They found, for instance, that the purine analogue 8-azaguanine has a very strong action in the chick and a much lesser one in the newt embryo. It is therefore necessary to consider the various classes of embryos separately. In this communication we shall be concerned only with chick embryos. Substances under test can be administered to such embryos by injection through the shell, as was done in the paper cited above With this technique it is impossible to know how much diffusion takes place of the substance injected, and one cannot therefore be certain of the effective concentration which actually reaches the embryo.

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Role of the chicken homeobox-containing genes GHox-4.6 and GHox-8 in the specification of positional identities during the development of normal and polydactylous chick limb buds

Development ◽

10.1242/dev.115.2.629 ◽

1992 ◽

Vol 115 (2) ◽

pp. 629-637 ◽

Cited By ~ 3

Author(s):

C.N. Coelho ◽

W.B. Upholt ◽

R.A. Kosher

Keyword(s):

Limb Development ◽

Ectopic Expression ◽

Limb Bud ◽

Chick Embryos ◽

Limb Buds ◽

Hox Gene ◽

Wing Bud ◽

Coated Bead ◽

Experimentally Induced

During early stages of normal chick limb development, the homeobox-containing (HOX) gene GHox-4.6 is expressed throughout the posterior mesoderm of the wing bud from which most of the skeletal elements including the digits will develop, whereas GHox-8 is expressed in the anterior limb bud mesoderm which will not give rise to skeletal elements. In the present study, we have examined the expression of GHox-4.6 and GHox-8 in the wing buds of two polydactylous mutant chick embryos, diplopodia-5 and talpid2, from which supernumerary digits develop from anterior limb mesoderm, and have also examined the expression of these genes in response to polarizing zone grafts and retinoic acid-coated bead implants which induce the formation of supernumerary digits from anterior limb mesoderm. We have found that the formation of supernumerary digits from the anterior mesoderm in mutant and experimentally induced polydactylous limb buds is preceded by the ectopic expression of GHox-4.6 in the anterior mesoderm and the coincident suppression of GHox-8 expression in the anterior mesoderm. These observations suggest that the anterior mesoderm of the polydactylous limb buds is “posteriorized” and support the suggestion that GHox-8 and GHox-4.6, respectively, are involved in specifying the anterior non-skeletal and posterior digit-forming regions of the limb bud. Although the anterior mesodermal domain of GHox-8 expression is severely impaired in the mutant and experimentally induced polydactylous limb buds, this gene is expressed by the prolonged, thickened apical ectodermal ridges of the polydactylous limb buds that extend along the distal anterior as well as the distal posterior mesoderm.(ABSTRACT TRUNCATED AT 250 WORDS)

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Correlation of wing-leg identity in ectopic FGF-induced chimeric limbs with the differential expression of chick Tbx5 and Tbx4

Development ◽

10.1242/dev.125.1.51 ◽

1998 ◽

Vol 125 (1) ◽

pp. 51-60 ◽

Cited By ~ 5

Author(s):

H. Ohuchi ◽

J. Takeuchi ◽

H. Yoshioka ◽

Y. Ishimaru ◽

K. Ogura ◽

...

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Expression Patterns ◽

Ectopic Expression ◽

Limb Bud ◽

Chick Embryos ◽

Fibroblast Growth ◽

Limb Buds ◽

Specific Position

It has been reported that members of the fibroblast growth factor (FGF) family can induce additional limb formation in the flank of chick embryos. The phenotype of the ectopic limb depends on the somite level at which it forms: limbs in the anterior flank resemble wings, whereas those in the posterior flank resemble legs. Ectopic limbs located in the mid-flank appear chimeric, possessing characteristics of both wings and legs; feather buds are present in the anterior halves with scales and claws in the posterior halves. To study the mechanisms underlying the chimerism of these additional limbs, we cloned chick Tbx5 and Tbx4 to use as forelimb and hindlimb markers and examined their expression patterns in FGF-induced limb buds. We found that Tbx5 and Tbx4 were differentially expressed in the anterior and posterior halves of additional limb buds in the mid-flank, respectively, consistent with the chimeric patterns of the integument. A boundary of Tbx5/Tbx4 exists in all ectopic limbs, indicating that the additional limbs are essentially chimeric, although the degree of chimerism is dependent on the position. The boundary of Tbx5/Tbx4 expression is not fixed at a specific position within the interlimb region, but dependent upon where FGF was applied. Since the ectopic expression patterns of Tbx5/Tbx4 in the additional limbs are closely correlated with the patterns of their chimeric phenotypes, it is likely that Tbx5 and Tbx4 expression in the limb bud is involved in determination of the forelimb and hindlimb identities, respectively, in vertebrates.

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The genesis of membrane bone in the embryonic chick maxilla: epithelial-mesenchymal tissue recombination studies

Development ◽

10.1242/dev.56.1.269 ◽

1980 ◽

Vol 56 (1) ◽

pp. 269-281

Author(s):

Mary S. Tyler ◽

David P. McCobb

Keyword(s):

Bone Formation ◽

Hind Limb ◽

Bone Grafts ◽

Chick Embryos ◽

Embryonic Chick ◽

Limb Buds ◽

Mesenchymal Tissue ◽

Membrane Bone ◽

Tissue Recombination

In the present study, the question of whether a relatively non-specific epithelial requirement exists for membrane bone formation within the maxillary mesenchyme was investigated. Organ rudiments from embryonic chicks of three to five days of incubation (HH 18–25) were enzymatically separated into the epithelial and mesenchymal components. Maxillarymesenchyme (from embryos HH 18–19) which in the absence of epithelium will not form bone was recombined with epithelium from maxillae of similarly aged embryos (homotypichomochronic recombination) and of older embryos (HH 25) (homotypic-heterochronicrecombination). Heterotypic recombinations were made between maxillary mesenchyme (HH 18–19) and the epithelium from wing and hind-limb buds (HH 19–22). Recombinants were grown as grafts on thechorioallantoic membranes of host chick embryos. Grafts of intact maxillae, isolated maxillary mesenchyme, and isolated epithelia from the maxilla, wing-, and hind-limb buds weregrown as controls. The histodifferentiation of grafted intact maxillae was similar to that in vivo; both cartilage and membrane bone differentiated within the mesenchyme. Grafts of maxillary mesenchyme (from embryos HH 18–19) grown in the absence of epithelium formed cartilage but did not form membrane bone. Grafts of maxillary mesenchyme (from embryos HH 18–19) recombined with epithelium in homotypichomochronic, homotypic-heterochronic, and heterotypic tissue combinations formed membrane bone in addition to cartilage. These results indicate that maxillary mesenchyme requires the presence of epithelium to promote osteogenesis and that this epithelial requirement is relatively non-specific in terms of type and age of epithelium.

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