Tbx16 cooperates with Wnt11 in assembling the zebrafish organizer.

The organizer, the signaling center that specifies vertebrate axial polarity and the nervous system, is a dorsal midline mesodermal domain in the gastrula that will form prechordal plate and anterior notochord. We show that in zebrafish the organizer is not a single domain when it first arises in the nascent mesoderm at the onset of gastrulation. Rather, in the presumptive prechordal plate region, the organizer is subdivided into two side-by-side cellular fields. Within minutes, concurrent medial and anterior cellular movements merge, or 'coalesce', the two fields to form the well-known singular midline field. Coalescence forms a symmetrical domain because the cell movements on the left and right sides initiate simultaneously and occur synchronously. However, in embryos with reduced function of the T-box transcription factor Tbx16 (Spadetail) or its genetic target paraxial protocadherin (Papc), synchrony is lost, coalesence is disrupted, and the midline domain is misshaped. Furthermore, with combined loss of Tbx16 and Wnt11 (Silberblick), coalesence is essentially absent. Possibly as a consequence, both the anterior movement of presumptive prechordal plate and organizer function, as assayed by eye-field separation, are disrupted. Our findings thus reveal that Tbx16, in combination with Wnt11, are critical components not only in morphogenesis but also in initial assembly of the organizer.