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. 2023 Jul 27;26(9):1516–1528. doi: 10.1038/s41593-023-01396-3

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© The Author(s) 2023, corrected publication 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a, Experimental strategy to unilaterally label Chx10-PPN neurons with an anterograde tracer (AAV-FLEX-tdTom-T2A-SypGFP) to reveal their projection pattern. b, Example fluorescence photomicrograph (coronal plane) of the rostral PPN at the injection site with tdTomato-expressing Chx10-PPN neurons in white. c, Reconstruction of putative synaptic bouton positions (digital SypGFP, cyan) from Chx10-PPN neuron projections at six different brainstem levels. The boutons are mostly located in the medial zone of the pontine and medullary reticular formations. Pie charts at each coronal level depict the percentage of boutons ipsilateral (ipsi, gray) or contralateral (contra, black) to the injection site, with the majority of them being ipsilateral at all brainstem levels. d, Normalized average bouton density in selected structures located within the coronal levels reconstructed in c. For each structure, the ipsilateral (gray) and contralateral (black) sides are quantified separately, unless they are midline structures (blue). Error bars represent the s.e.m. and hollow circles denote individual mice (N = 4 mice). For an extended overview and full names of all abbreviations, see Extended Data Fig. 10, which also includes projection areas in the midbrain and diencephalon.

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