Abstract
Enteric neural crest cells (ENCCs) colonize the gut during embryogenesis and migration defects give rise to Hirschsprung disease (HD). Mutations in GDNF/RET and EDN3/EDNRB are known to be causal in HD. Here, we show that migrating ENCCs in mice exhibit endogenous EDN3/EDNRB-gated calcium activity, mediated by chloride channels, T-type Ca2+ channels and inositol trisphosphate-sensitive intracellular-store release. We find that inhibiting Ca2+ activity results in ENCC migration defects, while exciting it promotes migration by increasing ENCC contractility and traction force to the extracellular matrix. Our study demonstrates that embryonic endothelin-mediated neural crest migration and adult endothelin-mediated vasoconstriction is one and the same phenomenon, taking place in different cell types. Our results suggest a functional link between rare mutations of CACNA1H (the gene encoding CaV3.2) and HD, and pave the way for understanding neurocristopathies in terms of neural crest cell bioelectric activity deficits.
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Essential data generated or analyzed during this study are included in the manuscript and supporting files. Source data are provided with this paper. Other non-essential data are available from the corresponding author upon request. Source data are provided with this paper.
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Essential codes for calcium imaging analysis are provided as supporting files.
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Acknowledgements
This manuscript is dedicated to Jürgen Langenbach, long-time science journalist at Die Presse, and to the memory of Prudence Dulormne. This research was funded by the Agence Nationale de la Recherche ANR GASTROMOVE - ANR-19-CE30-0016-01, by the Université de Paris IDEX Emergence en Recherche CHEVA19RDX-MEUP1, by the CNRS PEPS INSIS “COXHAM” grant, by the Labex “Who AM I?” ANR-11-LABX-0071, and by the Imaging platform BioEmergences-IBiSA, ANR-10-INBS-04 and ANR-11-EQPX-0029. We thank Sylvie Dufour for providing the Ht-PA::Cre mouse line, Ko Sugarawa for help with the Segment Anything Model under QuPath, Vincent Fleury, Michael Levin, Alexandre Ayed, Nathalie Rouach, Isabelle Arnoux, Olivier Romito and Master 2 students of the Université Paris Cité Biomedical Engineering 2024-2025 program for thoughtful discussions and/or performing experiments together.
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NRC led the project, obtained funding, performed experiments, analyzed data, synthesized data, wrote the draft and revised the paper; TS implemented new analysis methods and analyzed data; ZC performed experiments and analyzed data; NB, FG, MF, AEM, LC, MD, ILP, LZ performed experiments; LZ critically discussed the data; NB revised the draft.
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Chevalier, N.R., Gayda, F., Bondurand, N. et al. Endothelin-3 and T-type Ca2+ channels drive enteric neural crest cell calcium activity, contractility and migration. Nat Commun (2026). https://doi.org/10.1038/s41467-025-68121-5
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DOI: https://doi.org/10.1038/s41467-025-68121-5
