Expression of S9 and actin CyIIa mRNAs reveals dorso-ventral polarity and mesodermal sublineages in the vegetal plate of the sea urchin embryo

We have used whole mount in situ hybridization to analyze the patterns of expression of two genes, S9 and actin Cylla, during the development of the sea urchin, Strongylocentrotus purpuratus. We demonstrate that at the late blastula stage, these two mRNAs are expressed specifically by cells of the vegetal plate. Their domains of expression, however, are different. S9 mRNA is broadly distributed within most of the vegetal plate except for the central region, while Cylla expression is restricted to a population of 10-15 cells in the ventral region of the plate. S9-expressing secondary mesenchyme cells (SMCs) migrate from the vegetal plate into the blastocoel early in gastrulation and later populate the dorsal ectoderm. The numbers, morphology, and migratory behavior of these cells strongly suggest that they are pigment cells. Throughout gastrulation, Cylla mRNA is expressed by a population of presumptive SMCs at the ventral aspect of the archenteron tip. The pattern of expression of this mRNA is dynamic, however, and by the early pluteus stage, Cylla mRNA accumulates in primary mesenchyme cells (PMCs), SMCs, and endodermal cells of the gut. When embryos are treated with NiCl2, a compound that has been shown to ventralize other embryonic tissues, Cylla mRNA is expressed by an increased number of cells in the vegetal plate in a radially symmetrical pattern. The spatial pattern of Cylla expression provides the first direct molecular evidence that the vegetal plate is polarized along the dorso-ventral (D-V) axis of the embryo. This gene product should be a valuable marker in future studies of D-V axis specification, as it can be detected at earlier developmental stages than existing molecular markers of this axis. Our observations show that the vegetal plate consists of subterritories of gene expression, and provide further support for the view that diversification of the presumptive, non-skeletogenic mesoderm begins prior to the onset of invagination.