Geometric analysis of chondrogenic self-organisation of embryonic limb bud cells in micromass culture

Abstract. Insulin-like growth factors I and II have been shown differentially to affect the growth and carbohydrate metabolism of differentiating cartilage developed from mouse embryonic limb buds in organ culture. To gain insight into the relative importance of IGF-I and II actions in different stages of development of cartilage we have established a primary culture of differentiating chondrocytes from mouse embryonic limb buds. Trypsin digested limb bud cells from 9-11 day old mouse embryos differentiated into chondrocytes by 5-7 days in culture. At all stages of differentiation, distinct receptors of IGF-I and II were observed. IGF-I stimulated growth and sulphate incorporation of the non-differentiated and differentiated chondrocytes. IGF-II stimulated growth of the non-differentiated cells and had no effect on growth or sulphate incorporation by the differentiated cells. IGF-II, however, stimulated the glucose uptake by the cells at all stages of differentiation. These data confirm our previous suggestion that IGF-I in cartilage is the regulator of growth and differentiation, while IGF-II may be an important regulator of glucose metabolism in the tissue.

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Cell membrane and mitotic markers show that the neonatal rat gubernaculum grows in a similar way to an embryonic limb bud

Journal of Pediatric Surgery ◽

10.1016/j.jpedsurg.2007.04.035 ◽

2007 ◽

Vol 42 (9) ◽

pp. 1566-1573 ◽

Cited By ~ 14

Author(s):

Angelika F. Na ◽

Efrant J. Harnaen ◽

Pamela J. Farmer ◽

Magdy Sourial ◽

Bridget R. Southwell ◽

...

Keyword(s):

Cell Membrane ◽

Limb Bud ◽

Neonatal Rat ◽

Embryonic Limb

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NuSerum, a synthetic serum replacement, supports chondrogenesis of embryonic chick limb bud mesenchymal cells in micromass culture

In Vitro Cellular & Developmental Biology - Animal ◽

10.1007/bf02634229 ◽

1993 ◽

Vol 29 (12) ◽

pp. 917-922 ◽

Cited By ~ 19

Author(s):

Mayme Wong ◽

Rocky S. Tuan

Keyword(s):

Mesenchymal Cells ◽

Limb Bud ◽

Micromass Culture ◽

Embryonic Chick ◽

Serum Replacement ◽

Synthetic Serum

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Two distinct classes of prechondrogenic cell types in the embryonic limb bud

Developmental Biology ◽

10.1016/0012-1606(83)90063-5 ◽

1983 ◽

Vol 97 (1) ◽

pp. 59-69 ◽

Cited By ~ 46

Author(s):

B.J. Swalla ◽

E.M. Owens ◽

T.F. Linsenmayer ◽

M. Solursh

Keyword(s):

Cell Types ◽

Limb Bud ◽

Embryonic Limb

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SPECIFIC ECTODERMAL ENHANCERS CONTROL THE EXPRESSION OF Hoxc GENES IN DEVELOPING MAMMALIAN INTEGUMENTS

10.1101/2020.06.10.143677 ◽

2020 ◽

Author(s):

Marc Fernandez-Guerrero ◽

Nayuta Yakushiji-Kaminatsui ◽

Lucille Lopez-Delisle ◽

Sofía Zdral ◽

Fabrice Darbellay ◽

...

Keyword(s):

Gene Cluster ◽

Ectodermal Dysplasia ◽

Regulatory Elements ◽

Limb Bud ◽

Main Body ◽

Loss Of Function ◽

Causative Gene ◽

Embryonic Limb ◽

Mouse Limb ◽

Set Up

AbstractVertebrate Hox genes are key players in the establishment of structures during the development of the main body axis. Subsequently, they play important roles either in organizing secondary axial structures such as the appendages, or during homeostasis in postnatal stages and adulthood. Here we set up to analyze their elusive function in the ectodermal compartment, using the mouse limb bud as a model. We report that the HoxC gene cluster was globally co-opted to be transcribed in the distal limb ectoderm, where it is activated following the rule of temporal colinearity. These ectodermal cells subsequently produce various keratinized organs such as nails or claws. Accordingly, deletion of the HoxC cluster led to mice lacking nails (anonychia) and also hairs (alopecia), a condition stronger than the previously reported loss of function of Hoxc13, which is the causative gene of the ectodermal dysplasia 9 (ECTD9) in human patients. We further identified two ectodermal, mammalian-specific enhancers located upstream of the HoxC gene cluster, which act synergistically to regulate Hoxc gene expression in the hair and nail ectodermal organs. Deletion of these regulatory elements alone or in combination revealed a strong quantitative component in the regulation of Hoxc genes in the ectoderm, suggesting that these two enhancers may have evolved along with mammals to provide the level of HOXC proteins necessary for the full development of hairs and nails.Significance StatementIn this study, we report a unique and necessary function for the HoxC gene cluster in the development of some ectodermal organs, as illustrated both by the hair and nail phenotype displayed by mice lacking the Hoxc13 function and by the congenital anonychia (absence of nails) in full HoxC cluster mutants. We show that Hoxc genes are activated in a colinear manner in the embryonic limb ectoderm and are subsequently transcribed in developing nails and hairs. We identify two mammalian-specific enhancers located upstream of the HoxC cluster with and exclusive ectodermal specificity. Individual or combined enhancer deletions suggest that they act in combination to raise the transcription level of several Hoxc genes during hairs and nails development.

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BMPR-IA signaling is required for the formation of the apical ectodermal ridge and dorsal-ventral patterning of the limb

Development ◽

10.1242/dev.128.22.4449 ◽

2001 ◽

Vol 128 (22) ◽

pp. 4449-4461 ◽

Cited By ~ 7

Author(s):

Kyung Ahn ◽

Yuji Mishina ◽

Mark C. Hanks ◽

Richard R. Behringer ◽

E. Bryan Crenshaw

Keyword(s):

Limb Development ◽

Apical Ectodermal Ridge ◽

Bmp Signaling ◽

Conditional Knockout ◽

Limb Bud ◽

Gene Encoding ◽

Bone Morphogenetic Protein Receptor ◽

Protein Receptor ◽

Morphogenetic Protein ◽

Embryonic Limb

We demonstrate that signaling via the bone morphogenetic protein receptor IA (BMPR-IA) is required to establish two of the three cardinal axes of the limb: the proximal-distal axis and the dorsal-ventral axis. We generated a conditional knockout of the gene encoding BMPR-IA (Bmpr) that disrupted BMP signaling in the limb ectoderm. In the most severely affected embryos, this conditional mutation resulted in gross malformations of the limbs with complete agenesis of the hindlimbs. The proximal-distal axis is specified by the apical ectodermal ridge (AER), which forms from limb ectoderm at the distal tip of the embryonic limb bud. Analyses of the expression of molecular markers, such as Fgf8, demonstrate that formation of the AER was disrupted in the Bmpr mutants. Along the dorsal/ventral axis, loss of engrailed 1 (En1) expression in the non-ridge ectoderm of the mutants resulted in a dorsal transformation of the ventral limb structures. The expression pattern of Bmp4 and Bmp7 suggest that these growth factors play an instructive role in specifying dorsoventral pattern in the limb. This study demonstrates that BMPR-IA signaling plays a crucial role in AER formation and in the establishment of the dorsal/ventral patterning during limb development.

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