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Fig. 5 Schwann precursor–like cells are preserved in ret-/- zebrafish and are amenable to prucalopride-induced neurogenesis. (A) Graphical representation of gut isolation for scRNA-seq of 5-dpf tg(phox2bb:GFP) wild-type (WT) and ret-/- zebrafish. (B) UMAP visualization of WT and ret–/– zebrafish ENS subsets. (C) Live imaging of 5-dpf tg(phox2bb:kaede) photoconverted ret+/– zebrafish, treated with prucalopride. New phox2bb:kaede+ cells are indicated by asterisks (∗). (D) Quantification of new phox2bb:kaede+ cells in the proximal and (E) distal regions of DMSO (n = 8) and prucalopride (n = 9) treated 5-dpf photoconverted ret+/– tg(phox2bb:kaede) zebrafish. Statistical significance was calculated using Student t tests, ns > .05; ∗∗P < .01. (F) FISH combined with immunohistochemistry shows the presence of S100B+/RELN+/HTR4+ enteric glia located within hypertrophic nerve trunks in aganglionic colon (n = 3). In the insets, S100B staining is omitted for better appreciation of the FISH signal. IPAN, Intrinsic primary afferent neuron.