Three distinct lineages of mesenchymal cells in Ciona intestinalis embryos demonstrated by specific gene expression.

The ascidian embryonic mesenchyme, comprising about 900 cells, forms mesodermal tissues or organs of the adult body after metamorphosis. The mesenchyme originates from the A7.6 [trunk lateral cells (TLCs)], B7.7, and B8.5 blastomeres of the 110-cell stage embryo. Previous studies showed that FGF9/16/20 is required for specification of the mesenchyme in Ciona embryos and that two different (A7.6 and B8.5/B7.7) but partially overlapping molecular mechanisms are associated with the expression of a basic helix-loop-helix (bHLH) transcription factor gene, Twist-like1, in the mesenchymal precursors, which triggers the differentiation process of mesenchyme cells. In the present study, we examined whether the three embryonic lineages express the same mesenchyme-specific structural genes under the control of a common mechanism or whether the three lineages are characterized by the expression of genes specific to each of the lineages. We characterized nine mesenchyme-specific genes in Ciona embryos and found that five were expressed in A7.6/B8.5/B7.7, two in B8.5/B7.7, and two in B7.7 only. FGF9/16/20 and Twist-like1 were required for the expression of all the mesenchyme-specific genes, except for three A7.6/B8.5/B7.7-specific genes in A7.6 progenitors. Overexpression of FGF9/16/20 or Twist-like1 upregulated the expression of A7.6/B8.5/B7.7- and B8.5/B7.7-specific genes, while it downregulated the expression of B7.7-specific genes. These results provide evidence that the differentiation of each of the three mesenchyme lineages of Ciona embryos is characterized by the expression of a specific set of genes, whose expression is controlled differentially.