Abstract
Breathing is a bilaterally synchronous behavior that relies on a respiratory rhythm generator located in the brainstem. An essential component of this generator is the preBötzinger complex (preBötC), which paces inspirations. Little is known about the developmental origin of the interneuronal populations forming the preBötC oscillator network. We found that the homeobox gene Dbx1 controls the fate of glutamatergic interneurons required for preBötC rhythm generation in the mouse embryo. We also found that a conditional inactivation in Dbx1-derived cells of the roundabout homolog 3 (Robo3) gene, which is necessary for axonal midline crossing, resulted in left-right de-synchronization of the preBötC oscillator. Together, these findings identify Dbx1-derived interneurons as the core rhythmogenic elements of the preBötC oscillator and indicate that Robo3-dependent guidance signaling in these cells is required for bilaterally synchronous activity.
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Acknowledgements
We thank J.-F. Brunet, C. Goridis and A. Lumsden for comments on the manuscript; M. Tessier-Lavigne for providing the Robo3 knockout; S. Karaz for providing specimens; and S. Autran and V. Mézières for technical assistance with genotyping. J.B. is supported by Région Ile-de-France and the Fondation pour la Recherche Médicale. This work was supported by grants from Fondation pour la Recherche Médicale Equipe (A.C.), the Association Française contre les Myopathies (ASS-SUB06-00123, A.C.), the Ville de Paris (2006 ASES 102, A.P.) and the Agence Nationale de la Recherche (ANR-05-NEUR-007-01 BIS to A.P., ANR-08-MNPS-030-01 to A.C. and ANR-07-NEUR-007-01 to G.F.). This work benefited from the facilities and expertise of the Imagif Cell Biology Unit and the Anicampus mouse facility of the Gif-sur-Yvette campus. This work was supported by Centre National de la Recherche Scientifique and the Institut de la Santé et de la Recherche Médicale.
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J.E., A.P., J.C. and G.F. conceived the study. N.R. and A.C. designed the Robo3 experiments. J.B. and M.T.-B. performed the experiments. V.D. carried out Vglut2 in situ hybridization. J.B., M.T.-B. and G.F. analyzed the data. J.E., A.C. and A.P. provided reagents and mice. G.F. wrote the paper. All of the authors discussed the results and implications and commented on the manuscript at all stages.
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Split left/right breathing in Robo3 mutants. (MOV 735 kb)
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Bouvier, J., Thoby-Brisson, M., Renier, N. et al. Hindbrain interneurons and axon guidance signaling critical for breathing. Nat Neurosci 13, 1066–1074 (2010). https://doi.org/10.1038/nn.2622
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DOI: https://doi.org/10.1038/nn.2622
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