Neural plate pre-patterning enables specification of intermediate neural progenitors in the spinal cord.

Dorsal-ventral patterning of neural progenitors in the posterior neural tube, which gives rise to the spinal cord, has served as a model system to understand how extracellular signals organize developing tissues. While previous work has shown that signaling gradients diversify progenitor fates at the dorsal and ventral ends of the tissue, the basis of fate specification in intermediate regions has remained unclear. Here we use zebrafish to investigate the neural plate, which precedes neural tube formation, and show that its pre-patterning by a distinct signaling environment enables intermediate fate specification. Systematic spatial analysis of transcription factor (TF) expression and signaling activity using a reference-based mapping approach shows that the neural plate is partitioned into a striking complexity of TF co-expression states that, in part, correspond to the activity of gastrulation signals such as FGF and Wnt that persist through axis extension. Using analysis of cellular movement combined with fate mapping, we find that -expressing intermediate progenitors (p0) originate from a neural-plate specific state characterized by transient co-expression of the TFs , and . Finally, we show that this state is defined by Wnt signaling in the posterior neural plate and that ectopic Wnt activation within + cells is sufficient to promote expression. Our data broadly support a model in which neural progenitor specification occurs through the sequential use of multiple signals to progressively diversify the neural tissue as it develops. This has implications for differentiation of spinal cord cell types and for understanding signal-based patterning in other developmental contexts.