Cystatin B deficiency disrupts cerebellar redox homeostasis at 6 months of age
Redox assay | Cerebellum | ||
---|---|---|---|
Cystatin B+/+ | Cystatin B−/− | p value* | |
SODa (units/mg protein) | 632.1 ± 35.8 | 482.4 ± 20.6 | <0.05 |
GSHb (nmol/mg tissue) | 1.8 ± 0.004 | 1.0 ± 0.142 | <0.05 |
GSSG:GSHc | 0.061 ± 0.003 | 0.085 ± 0.006 | <0.05 |
8-epi PGF2αd (pg/mg) | 1.0 ± 0.09 | 5.6 ± 1.1 | <0.005 |
Glutathione peroxidasee (units/mg protein) | 24.4 ± 1.8 | 30.1 ± 1.3 | <0.05 |
↵*p values are determined by Student's t test. In the last column, p values corresponding to the following redox analyses are compared as follows.
↵aOne unit of SOD activity (units per milligrams of protein) is defined as 50% inhibition of formazan production. SOD activity was significantly reduced in Cystatin B−/− cerebella compared with wild-type controls [n = 5 (+/+), n = 4 (−/−)].
↵bGSH was significantly reduced in Cystatin B−/− cerebella compared with wild-type controls [n = 4 (+/+), n = 4 (−/−)].
↵cGSSG:GSH was significantly increased in Cystatin B−/− cerebella compared with wild-type controls [n = 4 (+/+), n = 4 (−/−)].
↵dLipid peroxidation was assayed based on competition between 8-epi–PGF2α and an 8-epi–PGF2α-acetylcholinesterase conjugate. Lipid peroxidation was significantly increased in Cystatin B−/− cerebella compared with wild-type controls [n = 3 (+/+), n = 5 (−/−)].
↵eGlutathione peroxidase activity was assayed by NADPH oxidation. Glutathione peroxidase activity increased significantly in Cystatin B−/− cerebella compared with wild-type controls [n = 3 (+/+), n = 5 (−/−)].