Additional file 2 of α-Synuclein induces prodromal symptoms of Parkinson’s disease via activating TLR2/MyD88/NF-κB pathway in Schwann cells of vagus nerve in a rat model
Creators: name: Cheng, Yue
name: Tong, Qing
name: Yuan, Yongsheng
name: Song, Xinna
name: Jiang, Wenwen
name: Wang, Yueping
name: Li, Wenjie
name: Li, Yangxia
name: Zhang, Kezhong
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Neuroscience
subject: Physiology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Science Policy
subject: Mental Health
subject: Computational Biology

description: Additional file 2: Figure S1. Vagal injection of AAV-A53T induces gastrointestinal dysfunction and subsequent motor deficits. A Representative image of stools collected in 12 h from rats among three groups at 3 months post-injection. Note that fecal pellets from AAV-A53T-injected rats are longer but less abundant. Latency to fall in the rotarod test (B), total distance (C) and average velocity (D), and representative images of the behavioral trajectories of rats in the open field test (E) at 3 months post-injection. Latency to fall in the rotarod test (F), total distance (G) and average velocity (H), and representative images of the behavioral trajectories of rats in the open field test (I) at 6 months post-injection. n = 12 per group. Data were presented as mean ± SEM. Statistical significance was analyzed using one-way ANOVA followed by Bonferroni’s multiple comparison test. ****P &lt; 0.0001. ns, not significant. Figure S2. Schematic diagram of electrophysiological testing of the vagus nerve. ①Vagus nerve; ② Stimulation electrode; ③ Recording electrode; ④ Recording electrode; ⑤ Ground electrode; ⑥ Ground electrode. Figure S3. Vagal injection of AAV-A53T induces aggregated α-synuclein deposited in the SCs of vagus nerves. A Representative confocal images of S100β (green), NF (violet), and MJF-14 (red) in the left vagus nerve at 3 months post-injection. Scale bar = 10 μm. B Representative confocal images of S129 (blue) and MJF-14 (red) in the left vagus nerve at 3 months post-injection. Scale bar = 10 μm. Figure S4. Vagal injection of AAV- Cas9-TLR2 induces a decline in TLR2 expression. Representative western blot bands (A) and statistical graph (B) of TLR2 in vagus nerves in rats among three groups at 3 months post-injection. The protein levels were normalized to GAPDH and expressed as fold-over control. n = 4. Data were presented as mean ± SEM. Statistical significance was analyzed using one-way ANOVA followed by Bonferroni’s multiple comparison test. ***P &lt; 0.001, ****P &lt; 0.0001. ns, not significant. Figure S5. TLR2 knockdown could partially reverse vagus nerve dysfunction. A colonic transit time. B 12 h fecal pellet count. C 12 h fecal total weight. D 12 h fecal water content. E Average intestinal blood flow. F Average vagus nerve conduction velocity. G Average amplitude of vagus nerve compound action potential. Counting numbers of non-myelinated axons (H) and myelinated axons (I) in the left vagus nerve at 3 months post-injection. A–D: n = 12 per group; E–I: n = 6 per group. Data were presented as mean ± SEM. Statistical significance was analyzed using Student’s t test. *P &lt; 0.05, ****P &lt; 0.0001, ns = not significant.