tph1 gene  (SAN GROUP)

Journal: Scientific Reports

Article Title: A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis

doi: 10.1038/s41598-024-57400-8

Figure Lengend Snippet: Systemic 3-IAld activates the Trp metabolism by enhancing the metabolic Tph1 peripheral pathway. ( A ) Mice were injected (i.p.) with 3-IAld or vehicle (Veh, DMSO 0.1% of olive oil) and Trp metabolites were quantified in serum and brain 1, 3 and 6 h after injection. ( B ) Trp metabolites and 3-IAld levels in murine serum and brain. Statistical analysis was performed using a Two-way ANOVA, Bonferroni post hoc test ( B ); (Vehicle, Veh (DMSO 0.1%)). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. Data represent the means ± SD from three independent experiments.

Article Snippet: All qPCR assays were performed using TaqMan probes (CYP1A1—Hs00153120_m1, TPH1—Hs00188220_m1) and TaqManTM Gene Expression Master Mix with a StepOneTM Plus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Injection

Journal: Scientific Reports

Article Title: A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis

doi: 10.1038/s41598-024-57400-8

Figure Lengend Snippet: 3-IAld activates the Trp metabolism by enhancing the metabolic Tph1 peripheral pathway in mast cells. ( A ) Cyp1a1 and Tph1 mRNA expression in WT and Ahr −/− BMDMCs. ( B ) 5-HT release in WT BMDMCs. ( C ) WT ex vivo sorted mast cells were analyzed for Tph1 mRNA expression ( D ) and Tph1 mRNA expression ( E ) in WT and Kit W /Kit W–v lymph node total cells. ( F ) Cyp1A1 and Tph1 mRNA expression in human CD34-derived mast cells. ( G ) Cytoplasmic and nuclear AhR translocation in WT BMDMCs exposed to 3-IAld or 3-IAld and DNP for 1 h. (Vehicle, Veh (DMSO 0.1%)). Statistical analysis was performed using Two-way ANOVA, Bonferroni post hoc test ( A , E ); Two-tailed Student’s t test, nonparametric Mann–Whitney U test ( B , D ); One-way ANOVA, Bonferroni post hoc test (B); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: All qPCR assays were performed using TaqMan probes (CYP1A1—Hs00153120_m1, TPH1—Hs00188220_m1) and TaqManTM Gene Expression Master Mix with a StepOneTM Plus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Expressing, Ex Vivo, Derivative Assay, Translocation Assay, Two Tailed Test, MANN-WHITNEY

Journal: Scientific Reports

Article Title: A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis

doi: 10.1038/s41598-024-57400-8

Figure Lengend Snippet: 3-IAld activates a mast cell tolerogenic program via AhR and Tph1. ( A ) Reactive Oxygen Species (ROS) detected in WT BMDMCs. ( B ) Real-time PCR and ELISA of the indicated targets in WT and Ahr −/− BMDMCs (Vehicle, Veh (DMSO 0.1%)). Statistical analysis was performed using Multiple T tests ( A ), Two-way ANOVA ( B ), Bonferroni post hoc test. n.s. , not significant. ∗ p < 0.05, ∗∗ p < 0.01, ∗ ∗ ∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: All qPCR assays were performed using TaqMan probes (CYP1A1—Hs00153120_m1, TPH1—Hs00188220_m1) and TaqManTM Gene Expression Master Mix with a StepOneTM Plus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis

doi: 10.1038/s41598-024-57400-8

Figure Lengend Snippet: 3-IAld systemic treatment protects mice from EAE inducing the AhR-Tph1-mast cell axis. ( A ) EAE was induced in WT, Ahr −/− , Tph1 −/− mice, 3-IAld (0.36 mg/mouse in olive oil) or Veh (DMSO 0.1% of olive oil) were injected i.p. at the indicated time points. ( B ) Mean clinical score registered every 2 days in WT, Ahr −/− , Tph1 −/− treated and untreated mice. ( C ) Cerebellum slice, viewed by transmission electron microscopy (scale bar = 5 µm) and hematoxylin and eosin stain on paraffin sections of spinal cords. Arrows indicate inflammatory infiltrates (scale bars = 100 μm). Assays were performed at 30 dpi and data were pooled from four independent experiments ( n = 8 mice per group); mean ± SEM. ( D ) Quantification of inflammatory infiltrates in WT, Ahr −/− , Tph1 −/− murine spinal cords with EAE and without EAE. Right panel, spinal cord hematoxylin and eosin paraffin section histology of non-EAE mice (scale bar = 50 μm). Statistical analysis was performed using a Multiple T test ( B ); Two-way ANOVA, Bonferroni post hoc test ( D ). ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: All qPCR assays were performed using TaqMan probes (CYP1A1—Hs00153120_m1, TPH1—Hs00188220_m1) and TaqManTM Gene Expression Master Mix with a StepOneTM Plus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Injection, Transmission Assay, Electron Microscopy, H&E Stain, Paraffin Section

Journal: Scientific Reports

Article Title: A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis

doi: 10.1038/s41598-024-57400-8

Figure Lengend Snippet: 3-IAld affects the cytokine microenvironment in peripheral lymph nodes via Tph1. 3-IAld is a tryptophan-derived indole metabolically released by several components of the gut microbiome. We have shown how the systemic administration of the indole-derivative activates the enzyme Tph1 in mast cells, in the lymph node compartments. The activation of the Tph1-mast cell axis is mediated by the binding of the nuclear receptor AhR. This mechanism supports how the administration of 3-IAld affects systemic inflammation in a mouse model of EAE and suggests that changes in serum concentration of indole-derivatives may affect the overall immunity.

Article Snippet: All qPCR assays were performed using TaqMan probes (CYP1A1—Hs00153120_m1, TPH1—Hs00188220_m1) and TaqManTM Gene Expression Master Mix with a StepOneTM Plus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Derivative Assay, Metabolic Labelling, Activation Assay, Binding Assay, Concentration Assay