Fig. 5

TrkA⁺ peripheral nerves express key angiogenic genes after soft tissue trauma. a–d Re-analysis of scRNA-Seq of mouse lumbar DRG neurons (8–12 weeks old male C57BL/6 J mice, n = 3 mice, 11 191 neuronal DRG cells were examined) (a) UMAP visualization of 4 cell clusters including Neurofilament (NF), Peptidergic nociceptors (PEP), Non-peptidergic (NP) and Tyrosine hydroxylase (TH) in pooled cells. b Violin plots demonstrating characteristic markers of each cell cluster. Nefh (Neurofilament Heavy Chain), Calca (Calcitonin Related Polypeptide Alpha), Tac1 (Tachykinin Precursor 1), Mrgprd (MAS Related GPR Family Member D), Ntrk1 (Neurotrophic Receptor Tyrosine Kinase 1), Th (Tyrosine Hydroxylase). c Heatmap of angiogenic gene expression among each cell cluster. Sema5a (Semaphorin 5 A), Fgf18 (Fibroblast Growth Factor 18), Fgf1 (Fibroblast Growth Factor 1), Vegfc (Vascular Endothelial Growth Factor C), Wnt5a (Wnt Family Member 5 A), Vegfa (Vascular Endothelial Growth Factor A), Pdgfa (Platelet Derived Growth Factor Subunit A), Vegfb (Vascular Endothelial Growth Factor B). d Module score analysis of angiogenic genes among each cell cluster. e–g qPCR analysis of angiogenic gene expression among ipsilateral lumbar DRGs after HO induction, in comparison to uninjured control. Ipsilateral L4-L6 pooled for analysis. e Schematic of qPCR analysis of angiogenic gene expression among ipsilateral lumbar DRGs after HO induction. Created in Biorender.com. f Heatmap of 15 pro-angiogenic genes enriched in Ntrk1-expressing neurons after HO induction, in comparison with uninjured control. Analysis performed at 3 d post-injury. g Representative most up-regulated genes upon HO induction, including Fgf18, Pdgfc, Vegfa, and Vegfd. h Representative immunofluorescent staining of ipsilateral L4 DRG after HO induction in relation to uninjured control, including FGF18 (Fibroblast Growth Factor 18), PDGFC (Platelet Derived Growth Factor Subunit C), VEGFA (Vascular Endothelial Growth Factor A), and VEGFD (Vascular Endothelial Growth Factor D). Analysis performed at 3 day post-injury, and quantification performed in relation to uninjured control. Scale bars: 100 μm. n = 3 or 4 animals per group. Individual dots in scatterplots represent values from single animal measurements, while mean and one SD are indicated by crosshairs and whiskers. * P < 0.05; ** P < 0.01. A two-tailed student’s t test was used (g, h). Non-parametric data was analyzed using the Kolmogorov–Smirnov test (d)