Fig. 5: Neuronal projections of submucosal classes within the mucosa.
a, Schematic drawing depicting non-ENS cell types/structures and their corresponding markers used in the study. Panel a is created with BioRender.com. b,c, Representative pictures showing TOM+ projections from the three smENCs around intestinal crypts (b) and inside villi (c). Arrowheads in insets in c indicate nerve fibers passing the basolateral side of goblet cells. d,e, Three-dimensional reconstructed Imaris images showing the spatial relation between the nerve fibers from smENCs and 5-HT+ EC cells (d) or DCLK+ tuft cells (e). See Supplementary Fig. 6 for original confocal pictures and for examples of long cell–cell distances. f–i, Analysis of the shortest distance between TOM+ nerve varicosity and EC cells or tuft cells using Imaris. f,h, Box-and-whisker plots showing individual shortest distances measured. Whiskers indicate the maximum and minimum values, boxes indicate the 25th–75th percentiles and the center line indicates the median. Number of cells analyzed from three mice of each line—51 (Nmu), 43 (Sst) and 60 (Vip) 5-HT+ EC cells; 44 (Nmu), 37 (Sst) and 64 (Vip) DCLK+ tuft cells. Each dot represents one cell. g,i, Density plots showing the distribution of the shortest distance measured. j, Aggregated circle plot showing number of interactions in CellChat analysis between smENCs and 15 epithelial cell types. k, Table showing examples of identified ligand–receptor pairs with epithelial cell types as senders and smENC1–smENC3 as receivers. l, Table showing examples of identified ligand–receptor pairs with smENC1–smENC3 as senders and epithelial cell types as receivers. Neurovascular spatial relation shown at three planes—submucosa (m), crypt (n) and villi (o). TOM+ fibers in all three Cre lines were most closely distributed to capillaries in the crypt and villi (Supplementary Videos 2–4). Representative images in b,c,m–o were based on observations in five fields of view from each mouse (n = 3). Scale bars, 20 µm (b,c,m–o). Mice age = 8–12 weeks. L, lymphatic vessel; A, arteriole; V, venule; EC, enterochromaffin; TA, transit amplifying.
