Fig. 6

Neuronal and glial loss induced by antibiotic (Abx) treatment are reversed by spontaneous microbial recolonization and are accompanied by enteric neurogenesis. a Representative immunofluorescent images of ganglia in the submucosal and myenteric plexuses: HuC/D⁺ (green) and nNOS⁺ (magenta) neurons in the ileum. Scale bar: 30 μm. b Number of HuC/D⁺ neurons in the submucosal (left panel) and myenteric plexus (middle panel) in whole-mount preparations of the ileum. The number of nNOS⁺ neurons in the myenteric plexus (right panel) in the ileum. c Number of HuC/D⁺ neurons in the submucosal (left panel) and myenteric plexus (middle panel) in whole-mount preparations of the colon. The number of nNOS⁺ neurons in the myenteric plexus (right panel) in the colon. d Representative immunofluorescent images of ganglia in the ileal myenteric plexus showing S100B⁺ enteric glial cells (EGC, green). Scale bar: 50 μm. e Relative number of S100B⁺ EGC in the ileal myenteric plexus. f Immunofluorescence images of representative ganglia of the colonic myenteric plexus: HuC/D⁺ (green) and Sox2⁺ (magenta) cells. Arrows represent double-labelled HuC/D⁺/Sox2⁺ neurons. Scale bar: 30 μm. g Number of HuC/D⁺ neurons (upper left panel), Sox2⁺ cells (lower panel), and double-labelled HuC/D⁺/Sox2⁺ neurons (upper right panel) in the myenteric plexus of the proximal colon. Data are expressed as mean ± SEM unless stated otherwise. a–c n = 3-5; d–g n = 5–10. *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA, followed by Tukey’s multiple comparison test