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Striated muscle cells in the tail of ascidian tadpole larvae differentiate cell-autonomously. Although several key regulatory factors have been identified, the genetic regulatory pathway has not fully been understood; comprehensive understanding of the regulatory pathway is essential for accurate modeling to deduce principles for gene regulatory network dynamics, and for comparative analysis how ascidians have evolved the cell-autonomous gene regulatory mechanism. Here, we revealed regulatory interactions among three key regulatory factors, Zic-r.b, Tbx6-r.b, and Mrf, and elucidated the mechanism by which these factors activate muscle structural genes. We revealed a cross-regulatory circuit among these regulatory factors, which maintained the expression of Tbx6-r.b and Mrf during gastrulation. While these two factors combinatorially activated muscle structural genes in late stage embryos, muscle structural genes were activated mainly by Tbx6-r.b before gastrulation. Time points when expression of muscle structural genes became first detectable were strongly correlated with the degree of Tbx6-r.b occupancy. Thus, the genetic pathway starting with Tbx6-r.b and Zic-r.b, which are activated by maternal factors and ending with expression of muscle structural genes, has been revealed.

The regulatory pathway from genes directly activated by maternal factors to muscle structural genes in ascidian embryos