Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: 5-HT promoted neurogenesis in the enteric nervous system. ( A ) The mRNA expression of 5-HT 3 , 5-HT 4 , and 5-HT 7 in the colon (n=6). ( B ) The protein expression of 5-HT 4 and GAPDH in the colon (n=3). ( C ) The mRNA expression of Tubb3, Sox2, Nestin, Olig2, and Gfap in the muscularis (n=6). ( D ) The protein expression of Tuj1, HUC/D, and GAPDH in the muscularis (n=3). ( E ) Immunofluorescence staining of HUC/D (green) and Tuj1 (red) in the LMMP (scale bar=100μm), and the number of HUC/D ( F ) and Tuj1 ( G ) positive cells was quantified. ( H ) Immunofluorescence staining of Tuj1 in the colon (Tuj1, red) and nucleic acid (Dapi, blue) (scale bar=100 μm), and ( I ) the number of Tuj1-positive cells was quantified. ( J ) Immunofluorescence staining of S100β in LMMP (S100β, red) and nucleic acid (Dapi, blue) (scale bar=100 μm), and ( K ) the number of S100β-positive cells was quantified. Data is presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: 5-HT 4 mediates 5-HT activation of the Wnt signaling pathway to promote neurogenesis. ( A and B ) The protein expression levels of β-catenin, APC, Axin1, GSK-3β, and GAPDH in the muscularis muscle were determined by Western blotting (n=3). ( C ) Western blotting was used to determine the protein expression of β-catenin, and GAPDH in N2a cells (n=3). ( D ) The expression of Tubb3, Sox2, and Nestin in N2a cells was measured by real-time qPCR (n=6). ( E – H ) The protein expression of β-catenin, HUC/D, Tuj1, and GAPDH in N2a cells was examined by Western blotting, and the relative protein levels were normalized to those of GAPDH (n=3). Data is presented as the mean ± SEM. * P < 0.05; ** P < 0.01.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: Overview of the role of 5-HT signaling in ELS-induced intestinal dysfunction. Elevated serotonin levels in the gut due to early stress contribute to intestinal dysfunction in mice. These effects may occur through activation of the 5-HT 4 receptor, stimulation of the Wnt pathway, the promotion of neurogenesis, an increase in excitatory neurons, and subsequent impacts on intestinal motility.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Activation Assay