Defect repair after somite removal in avian embryos is not true regeneration

Abstract

 The question of regeneration after experimental somite extirpation has been controversial in the literature. While all workers agree that repair of the defects occurs, results concerning the extent and mechanism of this process, as well as the origin of the cells filling the defect, show great discrepancies. Our approach towards a re-examination of this question involved microsurgical removal of individual somites in 2-day-chick embryos in combination with grafting of quail somites and lateral plate. We show that the defect in the paraxial mesoderm is filled within a day after extirpation and that the reconstituting cells are derived only from the cranial and caudal somites, but not from the contralateral somites or from the lateral plate. There are no indications of an increase of proliferation in the neighbouring somites. In order to examine the differentiation capacities of the cells that fill the defect, we used immunohistochemistry and in situ-hybridization. We show that the cells in the defect are mesenchymal in morphology and express Pax-1 and Twist. There are a few desmin-positive cells in the defect that can be shown to derive from adjacent somites. An epithelial dermomyotome and myotome are absent at the operation site. Neural crest cells do not participate in the reconstitution of the defect. We conclude that cells in the defect either already have or adopt a ventral somitic (sclerotomal) identity, whereas derivatives with dorsal identity are absent from the defect except for a few individual cells.