The temporal requirement for endothelin receptor-B signalling during neural crest development

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Abstract

Endothelin receptor B (EDNRB) is a G-protein-coupled receptor with seven transmembrane domains which is required for the development of melanocytes and enteric neurons. Mice that are homozygous for a null mutation in the Ednrb gene are almost completely white and die as juveniles from megacolon. To determine when EDNRB signalling is required during embryogenesis, we have exploited the tetracycline-inducible system to generate strains of mice in which the endogenous Ednrb locus is under the control of the tetracycline-dependant transactivators tTa or rtTA. By using this system to express Ednrb at different stages of embryogenesis, we have determined that EDNRB is required during a restricted period of neural crest development between embryonic days 10 and 12.5. Moreover, our results imply that EDNRB is required for the migration of both melanoblasts and enteric neuroblasts.

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Figure 1: Establishing tet-responsive alleles of Ednrb.
Figure 2: Kinetics of Ednrb regulation by dox.
Figure 3: The temporal requirement for Ednrb signalling.
Figure 4: Detection of melanoblasts and enteric neuroblasts by in situ hydridization.
Figure 5: Ednrb signalling is required during melanoblast migration.

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Acknowledgements

We thank L. Henninghausen and H. Bujard for providing constructs; H. Westphal for providing EIIa-cre transgenic mice; the members of our laboratory for helpful discussions; and M. Cleary for critical comments on the manuscript. M.K.S. was a Jane Coffin Childs Postdoctoral Fellow and S.M.T. is an Investigator of the Howard Hughes Medical Institute. R.S.I. isolated and characterized the Ednrb genomic clones and J.M.L. injected the targeted ES cells into blastocysts. M.K.S. and S.M.T. were responsible for the design of the experiment and the writing of the manuscript, and M.K.S. for the generation of the targeting vectors, targeting of ES cells and analysing the mice.

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Correspondence to Shirley M. Tilghman.

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Shin, M., Levorse, J., Ingram, R. et al. The temporal requirement for endothelin receptor-B signalling during neural crest development. Nature 402, 496–501 (1999) doi:10.1038/990040

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