Figure 7.

Oestrogen receptor β ligand treatment limits EAE-induced disorganization of nodal proteins in callosal axons. (A) Corpus callosum sections were immunostained with nodal proteins Caspr (red, marked with white arrows) and Nav1.6 (green). A significant decrease in Caspr and Nav1.6 staining occurred in the corpus callosum of vehicle-treated EAE mice. In addition, extensive regions of axons (*) were immunostained with Nav1.6 not confined between Caspr pairs. Oestrogen receptor β ligand-treated EAE corpus callosum axons contained Caspr pairs with Nav1.6 similar to normal control. Note: PLP-EGFP-green channel was dropped and Nav1.6 immunostaining performed with TRITC conjugated secondary was pseudo-coloured to green for clarity. (B) Quantification of Caspr protein pairs alone and Caspr protein pair encompassing Nav1.6 protein showed a significant decrease in vehicle-treated EAE callosal axons compared to those of normal and oestrogen receptor β ligand-treated EAE. **P < 0.001, ANOVAs; Bonferroni’s multiple comparison post test; n = 5 mice in each treatment group. (C) Juxtaparanodal Kv1.2 protein (red, arrows) immunostaining increased in the corpus callosum of vehicle-treated EAE mice. Specifically, Kv1.2 immunostaining was obvious throughout the length of some axons (*). No significant difference was observed in oestrogen receptor β ligand-treated EAE axons compared with normal.