The absence of ADAM11 protein in the (-/-) mice was confirmed by Western blot analysis. To be brief, the cerebellum was isolated from a six month-old mouse of each genotype and homogenised in cell lysis buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1% NP-40, protease inhibitor cocktail [Roche Diagnostics]). After removal of cell debris by centrifugation, the glycosylated proteins in the supernatant were concentrated by the affinity chromatography using Con A Sepharose 4B (Amersham Biosciences Corp.). Each sample was separated on 10% SDS-PAGE, and transferred to a PVDF membrane. The blot was then incubated with monoclonal antibody against the ADAM11 (U. S. Patent 5,631,351) at 1 μg/ml. Bound antibodies were visualised with horseradish peroxidase-labelled second antibody and an ECL-plus chemiluminescence detection system (Amersham Biosciences Corp.).