Fig. 1.

Characterisation of spinal cord slice cultures from neonatal PVG rat. Myelinated axons, indicated by β-III tubulin (green) and MBP (red) colocalisation, were not apparent in spinal cord slice cultures from P0–1 rat pups at 4 DIV (A) but were present at 7 DIV (B), scale = 50 µm for A, B. Immunohistochemical assessment of perfusion fixed spinal cord (C) and brain stem (D) from P0–1 rat pups showed no evidence of myelinated axons; myelinated axons were observed in spinal cord from P5 pups (E), scale = 50 µm for C–E. Electron microscopy of 7 DIV slice cultures revealed the presence of both relatively compact (F, G) and some decompacted myelin sheaths (H) in uncut slice cultures; cut injury 24 h previously resulted in increasingly decompacted myelin (I) and aggregates of myelin debris (J), scale = 1 µm. Immunoreactivity of 8OHdG (K) increased following cut injury (O), as did immunoreactivity of HNE (L, P), 3-NT (M, Q) and MnSOD (N, R). Immunoreactivity of all oxidative stress indicators is shown in red, together with MBP immunoreactivity (green) and Hoechst nuclear stain (blue); representative images adjacent to the cut injury are shown. Note that multiple colour immunofluorescence was employed and as such, 8OHDG and HNE images show the same field of view, as do 3NT and MnSOD images, scale = 50 µm