Zebrafish pigment cells develop directly from persistent highly multipotent progenitors

Neural crest cells are highly multipotent stem cells, but it remains unclear how their fate restriction to specific fates occurs. The direct fate restriction model hypothesises that migrating cells maintain full multipotency, whilst progressive fate restriction envisages fully multipotent cells transitioning to partially-restricted intermediates before committing to individual fates. Using zebrafish pigment cell development as a model, we show applying NanoString hybridization single cell transcriptional profiling and RNAscope in situ hybridization that neural crest cells retain broad multipotency throughout migration and even in post-migratory cells in vivo, with no evidence for partially-restricted intermediates. We find that leukocyte tyrosine kinase early expression marks a multipotent stage, with signalling driving iridophore differentiation through repression of fate-specific transcription factors for other fates. We reconcile the direct and progressive fate restriction models by proposing that pigment cell development occurs directly, but dynamically, from a highly multipotent state, consistent with our recently-proposed Cyclical Fate Restriction model.Published versionThis work was supported by Uehara Memorial Foundation (MN), Wellcome Trust VIP awards (M.N.), and BBSRC grants BB/ L00769X/1(R.N.K., H.S., T.S.) and BB/S015906/1 (R.N.K., J.H.P.D., K.C.S., G.B.) and BB/ L007789/1 and BB/S01604X/1 (A.R.), National Natural Science Foundation of China, Grant Number: 31000542 (X.Y.), Royal Society International Exchange Cost Share 2017 Russia award (R.N.K.), Russian Foundation of Basic Researcher grant 17-54-10014 (V.J.M.), Ministry of Science and Higher Education of the Russian Federation Grant number 075-15-2021-601 (V.J.M.), and University of Bath PhD Studentship and ORS award (T.J.C.)