Journal: Frontiers in Molecular Neuroscience
Article Title: C286, an orally available retinoic acid receptor β agonist drug, regulates multiple pathways to achieve spinal cord injury repair
doi: 10.3389/fnmol.2024.1411384
Figure Lengend Snippet: C286 induces functional recovery and tissue sparing in a model of spinal cord contusion. (A) C286 treatment leads to significant improvements in locomotor function in hind limbs in comparison to vehicle-treated rats. Data represent the mean ± SEM of n = 4, * p ≤ 0.01, ** p ≤ 0.005. “____” for the statistical significance of the right hind limbs and “___” for the left. One-way Analysis of Variance (ANOVA), followed by the Fisher test. (B) A schematic representation of the spinal cord area used for analysis of the injury pathology and cavity size. (C) Quantification of cavity area (expressed as a percentage of SC area) at 0 and 1 mm caudal to the epicentre at week 6 post-injury. Data represent the mean ± SEM, ** p ≤ 0.005, *** p ≤ 0.001, and Student's t -test. (D–I) C286 treatment leads to improved injury pathology and neuroprotection following a spinal contusion. Quantification of averaged (E) OX42, (F) NeuN, (H) GFAP, and (I) Iba1 expression shown as mean pixels in arbitrary units (a.u.) measured over a series of sections from 0–1 mm caudal to the epicentre revealed a significant preservation of spinal neurons and decreased reactive glia following C286 treatment compared with vehicle treatment. Data represent the mean FI ± SEM, ** p ≤ 0.005, *** p ≤ 0.001, and Student's t -test.
Article Snippet: They were incubated in the secondary antibody for 1 h at room temperature (RT) in PBS-0.02% Tween and then washed in PBS three times for 5 min. Antibodies used were: mouse monoclonal anti-βIII tubulin (Promega, 1:1,000 for immunohistochemistry), chicken polyclonal anti-GFAP (Abcam, 1:300); rabbit polyclonal anti-RARβ (Santa Cruz Biotechnology, Inc, 1:100 for immunohistochemistry), goat polyclonal anti-CGRP (Abcam, 1:200); mouse monoclonal anti-GAP-43 (Millipore, 1:20); mouse monoclonal anti-NF160 (Sigma Aldrich, 1:200); mouse monoclonal anti-OX42 (Abcam, 1:500); rabbit polyclonal anti-Iba1 (DAKO, 1:1,000); mouse monoclonal anti-CSPG (CS-56) (Sigma, 1:100), goat polyclonal anti-decorin (R&D Systems, 1:20, for immunochemistry and 5μg/ml as a neutralising antibody), rabbit polyclonal anti-decorin (Abcam, 1:50), mouse monoclonal anti-Osteopontin Invitrogen (ThermoFisher, 1:250); mouse monoclonal anti- integrin α9 (Abcam, 1:250); mouse monoclonal anti-tenascin-C (Abcam, 1:300).
Techniques: Functional Assay, Comparison, Expressing, Preserving