Figure 1. Cortical lesion model for initiating synaptic degeneration in the striatum of wild-type, heterozygous Wld^S and homozygous Wld^S mice.

A/B – Schematic diagram of the mouse brain viewed from above (A), showing the extent of cortical lesion produced (grey area). The dotted line in panel A represents the level of brain shown in coronal section in panel B (note the lesion to the left cortex). The box in panel B shows the region of striatum selected for ultrastructural experiments. C – Quantitative fluorescent western blotting of protein extracted from tail tips was used to confirm the genotype of experimental mice generated from heterozygous Wld^S (+/−) breeding colonies. Example blots show Wld^S protein levels (red; labelled with the Wld-18 antibody specific for Wld^S protein) and levels of actin loading control (green) in tail tips from 18 mice. 2 membranes are shown side by side with randomly arranged samples from individual mice numbered 1–18. A molecular weight marker is also shown (L). Lanes numbered 2,5,9,12 and 17 show wild-type mice (no Wld^S protein present), lanes 6,7,8,14,16 and 18 show heterozygous Wld^S mice (intermediate levels of Wld^S protein present), and lanes 1,3,4,10,11,13 and 15 show homozygous Wld^S mice (high levels of Wld^S protein present). D/E – Bar charts (mean±SEM) showing quantification of fluorescent western blots (see methods) shown in panel C (pooled to give a mean value for each genotype), confirming that heterozygous Wld^S mice had approximately half the expression levels of Wld^S protein observed in homozygous Wld^S mice (D), whilst levels of actin loading control remained constant (E) across mice of all genotypes. N = 5 wild-type mice, 6 heterozygous Wld^S mice & 7 homozygous Wld^S mice.