Cuprizone-Induced Demyelination and Recovery

All animals were fed a creatine-deficient amino acid diet (Envigo; TD.01084) unless otherwise specified. Demyelination was induced in 8-week-old male and female mice by adding 0.2% cuprizone (bis(cyclohexanone)oxaldihydrazone) into normal chow (LabDiet 5053) or creatine-deficient diet for 5 weeks. Cuprizone diet was replaced every 3 d to prevent stability concerns. All recovery diets used TD.01084 and added creatine or cyclocreatine (Sigma). Recovery diets after cuprizone were one of the following: normal chow, creatine-deficient diet, 2% creatine, or 0.1% cyclocreatine. Animals were weighed once every 2 d to ensure animals did not lose more than 10% body weight.