Reprogramming of cells during embryonic transfating: overcoming a reprogramming block.

Regulative development, demonstrated by many animal embryos, is the ability to replace missing cells or parts. The underlying molecular mechanism(s) of that ability is not well understood. If sea urchin micromeres (skeletogenic cell progenitors) are removed at the 16-cell stage, early endoderm initiates a sequential switch in cell fates, called transfating. Without micromeres, other mesoderm cells are absent as well, because their specification depends on signaling from micromeres. Most mesoderm cells later return by transfating, but pigment cells do not. Single-cell RNA sequencing, tracked over time, reveals the reprogramming sequence of those replacements. Beginning with an early endoderm specification state, cells progress through endomesoderm, then mesoderm, and finally distinct skeletogenic and blastocoelar cell specification states emerge, but pigment cells do not. Rescue of pigment cells was found to be a consequence of signal timing: if Delta is expressed prior to Nodal, pigment cells return. Thus, transfating operates through a series of gene regulatory state transitions, and reprogramming fails if endogenous negative signals occur prior to positive signals in the reprogramming sequence.