Fig. 5

Ex vivo and optogenetic activation of adrenergic downstream signaling triggers enteric glial reactivity. A Image frames taken during the acute exposure to isoprenaline (left panel, 10 µM) or Substance P (right panel, 10 µM) in Wnt1-Cre; R26R-GCaMP3^fl/fl mice. B Pie plot depicting the response types observed in the 14 ganglia (n = 4 mice). C Schematic of the primary culture preparation, viral transfection, and in vitro activation process of JellyOP^fll+ mice with blue light. D Confocal images of the JellyOP-GFP construct in tdTomato-Sox10⁺ cells seven days after transfection with the AAV-GFAP-Cre. Arrows indicate GFP⁺/GFAP⁺ glia. Scale bar (100 µm). E ELISA analysis for IL-6 and CCL2 (mean ± SEM) from conditioned medium of cultured primary EGCs from JellyOP^fl/+ mice transfected with an AAV-GFAP-Cre after stimulation with blue light or without stimulation (n = 24–42 separately transfected wells from two distinct isolations; Student's t-test, *** < 0.001). F Schematic of the in vivo activation process of Sox10^iCreERT2/Rpl22^HA/+/Ai14^fl/+/JellyOP^fl/+ mice. G Confocal images of IHC for FOS (green) and SOX10 (magenta) in whole mounts of Sox10^iCreERT2/Rpl22^HA/+/Ai14^fl/+/JellyOP^fl/+ or Sox10^iCreERT2/Rpl22^HA/+/Ai14^fl/+/JellyOP^+/+ mice 1 h after stimulation with blue light. Arrows indicate FOS-positive SOX10 glia. Scale bar (100 µm). H qPCR analysis of mRNA (mean ± SEM) from Sox10^iCreERT2/Rpl22^HA/+/Ai14^fl/+/JellyOP^fl/+l mice or Sox10^iCreERT2/Rpl22^HA/+/Ai14^fl/+/JellyOP^+/+ mice 3 h after optogenetic activation and laparotomy for gliosis hallmark genes (2^−ΔΔCT, 18S, JellyOP-negative animals, n = 3–7 animals per genotype; Student’s t-test, *** < 0.001, * < 0.05)