Journal: Journal of Neuroinflammation

Article Title: β-adrenergic signaling triggers enteric glial reactivity and acute enteric gliosis during surgery

doi: 10.1186/s12974-023-02937-0

Figure Lengend Snippet: Enteric glia react to mechanical stimuli and transition into an acute gliosis state. A Schematic description of the surgical procedure (intestinal manipulation, IM) with follow-up RiboTag approach in Sox10 iCreERT2 /Rpl22 HA/ + enteric glia and immunohistological image of HA (green) and SOX10 (magenta) co-expression in ME . Scale bar (100 µm). The RiboTag procedure was performed 3 h, 24 h, or 72 h after surgery. B Confocal images of SOX10 (magenta) and Ki67 (green) expression in whole mounts of small bowel ME at different time points. Scale bar (100 µm). C Histological analysis (mean ± SEM) of SOX10 + and SOX10 + Ki67 + cells per field of view (n = 6–14 animals per time point; mean counts of 5 images per n ± SEM; two-way ANOVA, to naive *** < 0.01, between IM24/IM72h ### < 0.01). D Principal component analysis (PCA) of a bulk RNA-Seq of Sox10 iCreERT2 /Rpl22 HA/ + RiboTag mRNA at different time points. E Volcano plot for actively transcribed genes at IM3h in Sox10 iCreERT2 /Rpl22 HA/ + enteric glia with significantly differentially transcribed genes (p-value < 0.05, > ± twofold) marked in red (upregulated) and blue (downregulated) and annotation of notable genes. F Venn diagrams of the top 50 induced genes at IM3h, IM24h, and IM72h separated into clusters. G Analysis of enriched GO-terms in mRNA from Sox10 iCreERT2 /Rpl22 HA/ + enteric glia for POI hallmarks related to migration and inflammatory response. H RNA-Seq heat maps for naïve and IM3h samples of Sox10 iCreERT2 /Rpl22 HA/ + RiboTag mRNA and total RNA “acute enteric gliosis” induction and an indication of genes related to key POI hallmarks. (n = 3–4 animals per time point)

Article Snippet: “Partek Flow” software was used to analyze RNA-Seq data (Lexogen pipeline 12,112,017), and Ensemble transcripts release 99 for mm10 mouse alignment.

Techniques: Expressing, RNA Sequencing Assay, Migration

Journal: Journal of Neuroinflammation

Article Title: β-adrenergic signaling triggers enteric glial reactivity and acute enteric gliosis during surgery

doi: 10.1186/s12974-023-02937-0

Figure Lengend Snippet: Acute enteric gliosis is modulated by sympathetic innervation. A Analysis of enriched GO-terms in mRNA from Sox10 iCreERT2 /Rpl22 HA/ + enteric glia 3 h after IM for POI hallmarks related to gliosis triggering pathways. B Heat map for the GO-term "GPCR signaling" in naïve and IM3h samples of mRNA from Sox10 iCreERT2 /Rpl22 HA/ + enteric glia and with selected genes highlighted. C Schematic description of chemical sympathetic denervation of C57BL6 mice with three consecutive intraperitoneal 6-OHDA injections (days 1–3). After 14 days, mice underwent IM. ME was isolated three hours later (IM3h) for qPCR and RNA sequencing. D Confocal images of immunohistological stainings of TUBB3 (magenta) and TH (green) expression in whole mounts of control (saline) and 6-OHDA treated (sympathectomized/STX) small bowel ME 17 days after injection. (n = 6 animals per condition). Scale bar (100 µm). E qPCR analysis showing fold changes of mRNA levels (mean ± SEM) from IM3h/Saline and IM3h/STX mice for enteric gliosis-related genes (2 −ΔΔCT , 18S , IM3h + saline; n = 6 animals per condition; Student’s t-test, * < 0.05, ** < 0.01). F Analysis of enriched POI hallmark GO-terms in IM3h/Saline total RNA and comparatively reduced in IM3h/STX samples related to inflammatory response and migration. G RNA-Seq heat map of our “acute enteric gliosis” GO-term for naïve and IM3h samples treated with saline or 6-OHDA (STX), and an indication of STX-affected genes (black line)

Article Snippet: “Partek Flow” software was used to analyze RNA-Seq data (Lexogen pipeline 12,112,017), and Ensemble transcripts release 99 for mm10 mouse alignment.

Techniques: Isolation, RNA Sequencing Assay, Expressing, Saline, Injection, Migration