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. 2019 Dec 10;9:18756. doi: 10.1038/s41598-019-55128-4

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© The Author(s) 2019

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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Intracolonic injection of DT results in focal aganglionosis. DT was injected into the mid-colon via laparotomy (a). Injection site is marked by India ink (a,b arrows). Immunofluorescence one week later shows focal loss of myenteric neurons (ablated area marked by a dotted line; c) by wholemount staining for Tuj1 (c,d) and Hu staining of a longitudinal section of mid-colon (e, arrows mark endogenous neurons at margins of DT ablation). One month after DT injection, focal ablation of Tuj1 + and Hu + enteric neurons (f,g) and S100β + enteric glia (h,i) was observed, with no obvious loss of SMA + smooth muscle (j,k) or c-kit + interstitial cells of Cajal (l,m). Scale bar 100 µm (c,d), 1 mm (e), and 100 µm (f–m).