Migration of myogenic cells from the somites to the fore‐limb buds of developing mouse embryos

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Abstract
In this study, we have isolated newly formed somites from the caudal regions of 8.5 day mouse embryos and transplanted them orthotopically into correspondingly staged hosts at the level of the prospective limb‐forming region. The experimental embryos were then cultured intact for 32‐36 hr. The donor somites used were pre‐labelled with DiI, a fluorescent lipophilic dye, or were obtained from transgenic embryos that carried a 1 kb 5′ regulatory sequence of the desmin gene linked to the gene encoding Escherichia coli β‐galactosidase. The transgene is specifically expressed in skeletal muscles (Li et al. [1993] Development 117:947‐959). The aim of these experiments was to show definitively that the musculature of the mammalian limb is derived from the somites. The results demonstrated that DiI‐labelled cells from the implanted somites were able to invade the proximal region of the fore‐limb bud during the course of development. The use of transgenic somites as grafts confirmed that some of the somitic cells found in the limbs were myogenic cells. To determine whether the displacement of somitic cells is an active or passive process, somatopleure obtained from the prospective limb‐forming regions of day 8.5 day embryos was implanted into 8.5 day hosts. We did not detect the presence of DiI‐labelled somatopleural cells in the fore‐limb after 32‐36 hr of culture. This suggests that somitic cells reached the limb bud via active locaomotion rather than as a result of being passively dragged there, as the limb elongates during development. In addition, we injected latex beads into the somites, as probes, to determine whether extracellular matrix‐driven translocation plays a role in driving the somitic cells to the limb bud. In a majority of the specimens examined, we could not detect the presence of these beads in the limb bud. However, in the trunk of these embryos, the beads were found dispersed throughout the ventral neural crest pathway.