Fig. 3

Enteric glia react before overt inflammation by receiving cues from the sympathetic nervous system. A Illustration of our hypothesis of TH⁺ neuron released NE triggering enteric glial activation and confocal images of immunohistological staining of sympathetic nerve fibers (TH, green), enteric neurons (MAP2; light blue), and enteric glia (GFAP, magenta). Arrows indicate TH⁺ fibers innervating the ME. Scale bar (50 µm). B Schematic description of chemical sympathetic denervation of Sox10^iCreERT2/Rpl22^HA/+ mice with three consecutive intraperitoneal 6-OHDA injections (days 1–3). After 14 days, the mice underwent Lap. ME was isolated three hours later (Lap3h), processed according to the RiboTag approach, and analyzed by qPCR. C Confocal images of immunohistological stainings of SOX10 (magenta) and FOS (green) expression in whole mounts of naïve and Lap3h small bowel ME. (n = 3 animals per condition). Arrows indicate FOS⁺ SOX10⁺ enteric glia. Scale bar (100 µm). D qPCR analysis showing fold changes of mRNA levels (mean ± SEM) of Sox10^iCreERT2/Rpl22^HA/+ RiboTag mRNA from naïve and Lap3h mice for cytokines (Ccl2, Il6) and an early response marker (Stat3) (2^−ΔΔCT, 18S/Tubb4, Naïve; n = 3 animals per condition; Student’s t-test, * < 0.05, ** < 0.01). (E) Confocal images of immunohistological stainings of TH (green) and endogenous SOX10-tdTomato (magenta) expression in whole mounts of Lap3h + Saline and Lap3h + STX small bowel ME. (n = 3 animals per condition). Scale bar (100 µm). F qPCR analysis showing fold changes of mRNA levels (mean ± SEM) of Sox10^iCreERT2/Rpl22^HA/+ RiboTag mRNA from Lap3h + Saline and Lap3h + STX mice for cytokines (Ccl2, Il6), and an early response marker (Stat3) (2^−ΔΔCT, Tubb4/Actb/PGK/GAPDH, Naïve; n = 3 animals per condition; Student’s t-test, * < 0.05, ** < 0.01)