Disruption of primary mesenchyme cell patterning by misregulated ectodermal expression of SpMsx in sea urchin embryos.

The patterning of the mesoderm of the sea urchin embryo is a classical paradigm of epithelial mesenchymal interactions in organogenesis, yet little is known of its molecular basis. Here we address the role of the homeobox gene, SpMsx, a member of the highly conserved Msx gene family, in this process. Msx genes have been shown to function in the dorsoventral patterning of the central nervous system in Drosophila and in a variety epithelial-mesenchymal interactions in vertebrates. We showed previously that the SpMsx gene is expressed during embryogenesis in a complex and dynamic pattern consistent with roles in the development of subpopulations of endoderm, mesoderm, and oral ectoderm. To perturb this pattern of expression and thus probe the function of SpMsx, we injected SpMsx mRNA into single-cell zygotes and monitored development morphologically and with a series of territory-specific molecular markers. RT-PCR analysis revealed that injected SpMsx transcripts persisted at least until the gastrula stage in amounts comparable to endogenous levels. Injected embryos exhibited deficiencies in the organization of primary and secondary mesenchyme cells within the blastocoelic cavity, as well as abnormalities in spicule number and shape. Defects in the endoderm were also common, including reduced or absent archenterons. Micromere transplantation experiments revealed that the defects in skeletogenic mesenchyme patterning were non-cell autonomous, consistent with findings that cell-cell interactions between ectoderm and the progenitors of the skeletogenic mesenchyme, the primary mesenchyme cells (PMCs), are important both for PMC guidance and spicule morphogenesis. Our data, taken together with observations in other organisms on the role of Msx genes in embryonic signaling processes, particularly involving the BMP pathway, suggest that SpMsx may be a part of the mechanism by which the ectoderm influences both the arrangement of primary mesenchyme cells within the blastocoel and the shapes of the skeletal rods.