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. 2022 May 27;15:832317. doi: 10.3389/fnmol.2022.832317

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Copyright © 2022 Natarajan, McCann, Dattani, Pachnis and Thapar.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Delayed migration of ENS progenitors along the gut of Ret^51/51 mutant embryos. (A,B) Immunostaining of representative cryosections through the foregut of control (TgRet^+/51) and TgRet^51/51 E9.5 embryos with GFP-specific antibodies. At this stage no difference was detected in the distribution of GFP-labelled vagal neural crest cells (nc) and pENCCs between control and mutant embryos. (C,D) GFP immunostaining of sections through the foregut of E10.5 control (TgRet^+/51) and TgRet^51/51 embryos. In control (TgRet^+/51) embryos, ENCCS are distributed throughout the foregut mesenchyme but in TgRet^51/51 mutants they fail to migrate much beyond the gastroesophageal junction. Insets show high power images of isolated GFP⁺ cells from control (TgRet^+/51) and mutant embryos, respectively. Arrowhead in inset (C) shows projections in control ENCC. (E,F) Whole mount preparations of gut dissected from control (TgRet^+/51) and TgRet^51/51 E11.5 embryos, immunostained for GFP showing that migration of TgRet^51/51 ENCCs is delayed relative to control TgRet^+/51 cells. (E’,F’) Sections through the foregut of control TgRet^+/51 and TgRet^51/51 embryos that correspond to the lines shown respectively in (E,F). (G,H) Whole mount preparations of gut dissected from control TgRet^+/51 and TgRet^51/51 E14.5 embryos immunostained for GFP. Control ENS progenitors have colonized the entire gut but Ret^51/51 progenitors have colonized only the foregut and the proximal half of the small intestine. Embryos from 3 litters per stage were used (approximately 4–6 per genotype). The embryos were isolated, genotyped individually and analyzed separately using immunolabelling. Scale bar: 200 μm. nc, neural crest; nt, neural tube; st, stomach; mg, midgut; c, caecum; hg, hindgut. TgRet^+/51 represents cells or embryos from genotype Wnt1^cre/+;R26R^stop/YFP;Ret^+/51 and TgRet^51/51 represents cells or embryos from genotype Wnt1^cre/+;R26R^stop/YFP;Ret^51/51.