title: Evidence that the limb bud ectoderm is required for survival of the underlying mesoderm.
abstract: The limb forms from a bud of mesoderm encased in a hull of ectoderm that grows out from the flank of the embryo. Coordinated signaling between the limb mesoderm and ectoderm is critical for normal limb outgrowth and patterning. The apical ectodermal ridge (AER), found at the distal tip, is a rich source of signaling molecules and has been proposed to specify distal structures and maintain the survival of cells in the underlying distal mesoderm. The dorsal and ventral non-AER ectoderm is also a source of signaling molecules and is important for dorsal-ventral patterning of the limb bud. Here we determine if this ectoderm provides cell survival signals by surgically removing the dorsal or ventral ectoderm during early chicken limb bud development and assaying for programmed cell death. We find that, similar to the AER, removal of the dorsal or ventral non-AER ectoderm results in massive cell death in the underlying mesoderm. In addition, although a re-epithelialization occurs, we find perturbations in the timing of Shh expression and, for the case of the dorsal ectoderm removal, defects in soft tissue and skeletal development along the proximal-distal axis. Furthermore, ectoderm substitution experiments show that the survival signal produced by the dorsal limb ectoderm is specific. Thus, our results argue that the non-AER ectoderm, like the AER, provides a specific survival signal to the underlying mesoderm that is necessary for normal limb development and conclusions drawn from experiments in which the non-AER ectoderm is removed, need to take into consideration this observation.
authors.affiliation: Departamento de Anatomía y Biología Celular, Facultad de Medicina, Universidad de Cantabria, 39011 Santander, Spain.
authors.forename: Marian
authors.initials: M
authors.lastname: Fernandez-Teran
authors.forename: Maria A
authors.initials: MA
authors.lastname: Ros
authors.forename: Francesca V
authors.initials: FV
authors.lastname: Mariani
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000254:growth & development / Q000378:metabolism; D019521:Body Patterning; D016923:Cell Death; D002470:Cell Survival; D002642:Chick Embryo; D002645:Chickens / Q000254:growth & development* / Q000378:metabolism; D004475:Ectoderm / Q000166:cytology / Q000378:metabolism*; D051524:Fibroblast Growth Factor 8 / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D053823:Hedgehog Proteins / Q000235:genetics / Q000378:metabolism; D018878:Limb Buds / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism*; D013997:Time Factors; D014921:Wings, Animal / Q000196:embryology
chemical_list: D053823:Hedgehog Proteins; D051524:Fibroblast Growth Factor 8
keywords: AER; Cell death; Dorsal ectoderm; Limb development; Pattern formation