the regenerative plasticity of isolated urodele myofibers and its dependence on msx1孤立urodele肌纤维再生可塑性及其msx1的依赖.pdfVIP

Open access, freely available online PLoS BIOLOGY The Regenerative Plasticity of Isolated Urodele Myofibers and Its Dependence on Msx1 1 2 3 1* Anoop Kumar , Cristiana P. Velloso , Yutaka Imokawa , Jeremy P. Brockes 1 Department of Biochemistry and Molecular Biology, University College London, London, United Kingdom 2 Department of Anatomy and Developmental Biology, Royal Free and University College Medical School, London, United Kingdom 3 Center for Developmental Biology, Laboratory for Evolutionary Regeneration, RIKEN, Chuo-ku, Kobe, Japan The conversion of multinucleate postmitotic muscle fibers to dividing mononucleate progeny cells (cellularisation) occurs during limb regeneration in salamanders, but the cellular events and molecular regulation underlying this remarkable process are not understood. The homeobox gene Msx1 has been studied as an antagonist of muscle differentiation, and its expression in cultured mouse myotubes induces about 5% of the cells to undergo cellularisation and viable fragmentation, but its relevance for the endogenous programme of salamander regeneration is unknown. We dissociated muscle fibers from the limb of larval salamanders and plated them in culture. Most of the fibers were activated by dissociation to mobilise their nuclei and undergo cellularisation or breakage into viable multinucleate fragments. This was followed by microinjection of a lineage tracer into single fibers and analysis of the labelled progeny cells, as well as by time-lapse microscopy. The fibers showing morphological plasticity selectively expressed Msx1 mRNA and protein. The uptake of morpholino antisense oligonucleotides directed to Msx1 led to a specific decrease in expression of Msx1 protein in myonuclei and marked inhibiti