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. 2024 Oct 25;15:9237. doi: 10.1038/s41467-024-53545-2

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — A scRNA-seq uniform manifold approximation and projection (UMAP) of Tph2⁺ enteric neurons from the small intestine of infant mice. Left, subtypes of enteric neurons according to ref. ⁶¹; PSN, putative sensory neurons; PIN, putative inhibitory neuron; PIMN, putative inhibitory motor neurons; PEMN, putative excitatory motor neurons; PSVN, putative secretomotor/vasodilator neurons were defined according to⁶¹. Right, distribution of Tph2 transcripts among subsets of enteric neurons. B Detection of Tph2, Hand2, and Tph1 in bulk RNA-seq analysis of the intestinal epithelium, lamina propria immune cells, and enteric neurons (n = 9 mice); TPM, Transcripts Per Million. C Representative whole-mount staining of small intestine from Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice. Neurons are detected with Tuj1 (red) and 5-HT positive cells with anti-serotonin antibody (green). Yellow arrows point to enteric serotonergic neurons (round-shaped cells) in the lamina propria, where serotonin co-localizes with Tuj1. The triangular-shaped serotonin positive cells observed in both Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice likely represent enterochromaffin cells. Scale bar, 100 μm. Similar results were obtained in at least 5 animals. D Fecal IgA levels in Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice (n = 21). E Representative immunostaining of small intestine sections from Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice. Epithelial cells are detected with EpCAM (magenta), IgA cells with anti-IgA antibody (green), and nuclei with DAPI (blue). Scale bar, 100 μm. F Flow cytometry analyses of frequency of total B cells, IgD⁺ fraction of total B cells, and IgA⁺ cells in the lamina propria CD45⁺ cells from the small intestine of Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice (n = 9). G STm CFU burdens in spleens and livers of Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice 5 days post oral inoculation (n = 12). H Kaplan–Meier plot of survival of Tph2^fl/fl and Tph2^fl/fl; Hand2-Cre mice following oral STm inoculation (n = 16). Data shown are means ± SD. Statistical analysis was performed by two-tailed Mann–Whitney test in D, F, G; and by a Log-rank test in H.