compound 21 11-(1-piperazinyl)-5h-dibenzo [b, e] [1, 4] diazepine abbreviated as c21  (Tocris)

Journal: Cell reports

Article Title: Peripheral neuronal activation shapes the microbiome and alters gut physiology

doi: 10.1016/j.celrep.2024.113953

Figure Lengend Snippet: (A) Experimental paradigm. Cre-dependent hM3Dq was virally administered to either ChAT-Cre or TH-Cre mice. After 3–4 weeks of expression, C21 was injected daily for 10 days to induce specific neuronal activation. Feces was sampled the day prior to the first C21 injection and on days 2, 6, and 10 of C21 administration, and tissue and cecal contents were collected 1 h after the last injection. (B) Faith’s phylogenetic diversity of feces and cecal contents over 10 days of neuronal activation in ChAT + and TH + mice. Feces was collected pre-experiment (1 day before the first injection) and on days 2, 6, and 10. Cecal contents were collected at the experimental endpoint on day 10. **p < 0.01, ***p < 0.001, determined by Kruskal-Wallis one-way ANOVA. (C–H) Weighted UniFrac principal-coordinate analysis (PCoA) of activated vs. control in ChAT + and TH + mice. Statistics were performed in QIIME2 as in Bolyen et al. (I) Stacked bar graph showing phylum-level changes in relative abundance on day 6 and day 10 of injection for feces and day 10 for cecal contents. (J–M) Linear discriminant analysis effect size (LEfSe) of the cecal microbiome. Cladograms show differential phylogenetic clusters and family-level differences in activated and control (J and K) ChAT + and (L and M) TH + mice. Cutoff: log 10 (LDA score) > 2 or < −2. (N and O) Changes in relative abundance of A. muciniphila in feces and cecal contents of (N) ChAT + and (O) TH + mice (n = 11–14 mice per group per time point). Red, control; green, hM3Dq-activated. *p < 0.05, **p < 0.01, determined by multiple t tests with Holm-Sidak correction for multiple comparisons. (P–S) Beta diversity of bacterial gene families and pathways in the (P and R) cecum and (Q and S) post-9 feces of control and activated mice. The direction and length of arrows indicate their influence in separating control and activated groups. Colors represent gene families and pathways (annotated in ). See also and . Source can be found at https://github.com/mazmanianlab/Griffiths_Yoo_et_al/tree/main/metagenomics .

Article Snippet: Compound 21 dihydrochloride (C21) , HelloBio , Cat# HB6124.

Techniques: Expressing, Injection, Activation Assay, Control

Journal: Cell reports

Article Title: Peripheral neuronal activation shapes the microbiome and alters gut physiology

doi: 10.1016/j.celrep.2024.113953

Figure Lengend Snippet: (A) Volcano plot of differentially expressed host proteins identified in the cecal contents of ChAT + -activated (n = 8) vs. ChAT + control (n = 9) mice 1 h after final C21 administration. (B) STRING network analysis of host proteins that were more abundant in ChAT + -activated mice (p nom. < 0.2). (C) Proteomics volcano plot of TH + -activated vs. TH + control mice (n = 7 mice per group). (D) STRING network analysis of upregulated host proteins in TH + -activated mice (p nom. < 0.2). (E and F) Unipept metaproteomics analysis of upregulated microbial proteins (fold change > 2, p nom. < 0.2) in TH + -activated and ChAT + -activated mice (n = 7–9 mice per group). Source data for (A) can be found at https://github.com/mazmanianlab/Griffiths_Yoo_et_al/blob/main/proteomics/CHAT_proteomics_volcano.txt . Source data for (C) can be found at https://github.com/mazmanianlab/Griffiths_Yoo_et_al/blob/main/proteomics/TH_proteomics_volcano.txt . Source data for (E) and (F) can be found at https://github.com/mazmanianlab/Griffiths_Yoo_et_al/blob/main/proteomics/metaproteomics/Microbiome_associated_proteins.xlsx .

Article Snippet: Compound 21 dihydrochloride (C21) , HelloBio , Cat# HB6124.

Techniques: Control, Metaproteomics

Journal: Cell reports

Article Title: Peripheral neuronal activation shapes the microbiome and alters gut physiology

doi: 10.1016/j.celrep.2024.113953

Figure Lengend Snippet: (A) Activation-mediated changes in whole-gut transit time in ChAT + and TH + mice. (B) Activation-mediated changes in fecal pellet output in ChAT + and TH + mice. (C) Activation-mediated changes in normalized cecal content mass in ChAT + and TH + mice. (D and E) Fecal pellet water content in (D) ChAT + and (E) TH + mice over 2 h following C21 activation, with a least-squares nonlinear regression displaying a 95% confidence interval. (F) Average fecal pellet water content in ChAT + and TH + mice following activation. (A–F) (n = 10–11 mice per group). *p < 0.05, ***p < 0.001, ****p < 0.0001, determined by 2-way ANOVA with Sidak′s method for multiple comparisons. (G and H) Frequency of ex vivo CMMCs from (G) ChAT + and (H) TH + mice over 30 min following activation (n = 3–6 mice per group). **p < 0.01, ***p < 0.001, determined by 2-way ANOVA with Sidak′s method for multiple comparisons. (I and J) Heatmaps showing frequency of CMMCs over 400 s following activation in ex vivo preparations from (I) ChAT + control and (J) ChAT + DREADD-administered mice. See also and .

Article Snippet: Compound 21 dihydrochloride (C21) , HelloBio , Cat# HB6124.

Techniques: Activation Assay, Ex Vivo, Control

Journal: Cell reports

Article Title: Peripheral neuronal activation shapes the microbiome and alters gut physiology

doi: 10.1016/j.celrep.2024.113953

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Compound 21 dihydrochloride (C21) , HelloBio , Cat# HB6124.

Techniques: Recombinant, Fluorescence, Plasmid Preparation, Software