Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Targeted muscle reinnervation attenuates neuropathic pain and neuroma development in a rat model of tibial nerve transection

doi: 10.3389/fbioe.2026.1758496

Figure Lengend Snippet: Immunohistochemical analysis of the distal segment of the tibial nerve. (A) Immunohistochemical expression of c-Fos, α-SMA, and Substance-P in longitudinal sections of the distal tibial nerve from each group. Scale bar = 100 μm. (B–D) Quantitative analysis of the positive area for c-Fos, α-SMA, and Substance-P detected by immunohistochemistry. Data are expressed as means ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: no significance. n = 5 for all groups.

Article Snippet: For DRG sections, primary antibody incubation was similarly performed overnight at 4 °C with anti-c-Fos (Rabbit, 1:100, Affinity, AF5354), anti-α-Smooth Muscle Actin (α-SMA) antibody (Rabbit, 1:3000, Proteintech, 14395-1-AP), and anti-Substance P antibody (Rabbit, 1:100, Bioss, bs-0065R).

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Mast Cells Inhibit Stem Cell-driven Epithelial Repair in Inflammatory Bowel Disease via Suppressing Wnt/lrp6/β-Catenin Signaling Pathway

doi: 10.1016/j.jcmgh.2026.101741

Figure Lengend Snippet: In vitro analysis of activated MCs’ direct inhibitory/cytotoxic effects on mouse intestinal organoids, and reversal by Wnt agonists. ( A – I ) Activated BMMCs’ effects on organoids (BMMC− group: organoid cocultured without active BMMC; BMMC+ group: organoid cocultured with active BMMC) ( A ) Co-culture schematic: Activated BMMCs and organoids were co-cultured to assess direct interactions. ( B and C ) Organoid growth: ( B ) Bright-field images; ( C ) quantification of organoid size, bud number, and density. ( D–G ) qRT-PCR analysis: ( D ) ISC markers, ( E ) stemness genes, ( F ) apoptosis genes, ( G ) Wnt pathway genes. ( H and I ) Representative images showing IF staining: Staining for total β-catenin ( H ) and active β-catenin ( I ). (n = 6–10 organoids/group). ( J–Q ) Wnt agonists reverse activated BMMC-induced inhibition (Vehicle group: organoid with DMSO and Substance P; BMMC+DMSO group: organoid cocultured with activated BMMC and treated with DMSO; BMMC+HLY78 group: organoid cocultured with activated BMMC and treated with Wnt agonist HLY78), ( J and K ) Organoid growth recovery: ( J ) Bright-field images; ( K ) quantification of size, bud number, density. ( L–O ) qRT-PCR analysis: ( L ) ISC markers, ( M ) stemness genes, ( N ) Wnt pathway genes, ( O ) apoptosis genes. ( P and Q ) Representative images showing IF staining: Staining for total β-catenin ( P ) and active β-catenin ( Q ). (n = 4–6 organoids/group). Statistical significance: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: To mimic the pathological environment of colitis, MCs were activated with 50 μM substance P (MCE, HY-P0201), based on the established role of substance P-dependent MC activation in colitis.

Techniques: In Vitro, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Staining, Inhibition

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Mast Cells Inhibit Stem Cell-driven Epithelial Repair in Inflammatory Bowel Disease via Suppressing Wnt/lrp6/β-Catenin Signaling Pathway

doi: 10.1016/j.jcmgh.2026.101741

Figure Lengend Snippet: The effect of CPA inhibitor on MC-induced inhibition of ISC-mediated epithelial regeneration. Four groups were set: Control (organoids + Substance P + DMSO), CPA inhibitor (organoids + Substance P + CPA inhibitor), BMMC (organoids + activated BMMCs + DMSO), BMMC + CPA inhibitor (organoids + activated BMMCs + CPA inhibitor). All co-cultures lasted 3 days before analysis. ( A and B ) Organoid growth phenotypes after CPA inhibitor intervention. ( A ) Bright-field microscopy images showing organoid morphology in the 4 groups. ( B ) Quantitative analysis of organoid size, bud number, and density. ( C–F ) Gene expression analysis via qRT-PCR. Relative mRNA levels of ISC marker genes ( C ), stemness-associated genes ( D ), Wnt pathway genes ( E ), and apoptosis-related genes ( F ). ( G and H ) Representative images showing IF staining of total β-catenin ( G ) and active β-catenin ( H ) in colon (n = 4–6 organoids/group). Statistical significance: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: To mimic the pathological environment of colitis, MCs were activated with 50 μM substance P (MCE, HY-P0201), based on the established role of substance P-dependent MC activation in colitis.

Techniques: Inhibition, Control, Microscopy, Gene Expression, Quantitative RT-PCR, Marker, Staining

Journal: Frontiers in Physiology

Article Title: Plasticity of TRPV1-Expressing Sensory Neurons Mediating Autonomic Dysreflexia Following Spinal Cord Injury

doi: 10.3389/fphys.2012.00257

Figure Lengend Snippet: High-thoracic (T3) spinal cord injury-induced selective hypertrophy of sensory neurons expressing the capsaicin receptor (TRPV1) and the artemin receptor (GFRα3) in the L4/L5 DRG . (A) Substance P (SP) – positive DRG neurons and their size-frequency distributions. (B) IB4-binding DRG neurons. (C) P2X3-positive DRG neurons. (D) TRPV1-positive DRG neurons. (E) GFRα3-expressing DRG neurons, known to express TRPV1. The overall proportions of immunopositive neurons [insets (A–E) ] did not change for any subpopulation. Ganglia were harvested 3 months after sham-injury (gray) or complete T3 SCI (black). Scale bar = 50 μm. Asterisks indicate P < 0.05, K–S goodness-of-fit test.

Article Snippet: For immunohistochemistry, slides were incubated in 10% normal donkey serum in PBS plus Triton X-100 (0.1%) for 20 min. We used five antibodies to delineate different subsets of nociceptors: these included antibodies raised against TRPV1 (Neuromics, Edina, MN, USA; 1:2,000), the ionotropic ATP purinoceptor P2X 3 (Millipore, Billerica, MA, USA; 1:1,000), Substance P (Neuromics, 1:1,000), the glycoprotein isolectin B4 (IB4; Neuromics; 1:2,000), the glial cell line-derived neurotrophic factor family receptor α 3 (GFRα3; R&D Systems; 1:500).

Techniques: Expressing, Binding Assay