Conservation of the developmental role of Brachyury in notochord formation in a urochordate, the ascidian Balocynthia roretzi

The notochord is one of the characteristic features of the phylum Chordata. The vertebrate Brachyury gene is known to be essential for the terminal differentiation of chordamesoderm into notochord. In the ascidian, which belongs to the subphylum Urochordata, differentiation of notochord cells is induced at the late phase of the 32-cell stage through cellular interaction with adjacent endoderm cells as well as neighboring notochord cells. The ascidian Brachyury gene (As-T) is expressed exclusively in the notochord-lineage blastomeres, and the timing of gene expression at the 64-cell stage precisely coincides with that of the developmental fate restriction of the blastomeres. In addition, experimental studies have demonstrated a close relationship between the inductive events and As-T expression. In the present study, we show that overexpression of As-T by microinjection of the synthesized As-T RNA results in the occurrence, without the induction, of notochord-specific features in the A-line presumptive notochord blastomeres. We also show that overexpression of As-T RNA leads to ectopic expression of notochord-specific features in non-notochord lineages, including those of spinal cord and endoderm. These results strongly suggest that the developmental role of the Brachyury is conserved throughout chordates in notochord formation.