Cuprizone bis cyclohexanone oxaldihydrazone

Adult (C57BL/6, eight-week-old, male) mice were purchased form the animal research center of shanghai laboratory and were housed at 22°C-24°C. Food and water were available ad libitum. Animals were cared for in accordance with the National Institutes of Health Guidelines for Animal Care. All experimental procedures were approved by Animal Care and Use Committee in Southeast University.
At the beginning of the research, the mice were randomly divided into three groups. Three groups included the control group (n = 17), the non-pharmacologic treatment Cuprizine-fed group (n = 17) and the pharmacologic treatment Cuprizine-fed group (n = 17). 8-Week-old C57/Bl6 mice were fed with 0.2% (w/w) cuprizone (bis-cyclohexanone oxaldihydrazone) (Sigma) in ground breeder chow for about ten weeks.

Total 6 male Sprague-Dawley rats with 8-9 weeks of age were scanned in this study approved by the Institutional Animal Care and Use Committee at the Korea Basic Science Institute. Before MRI scan, rats were initially anesthetized by 5% isoflurane mixed with a 3 : 7 mixture of pure oxygen and nitrous oxide in a plastic box. The animals were then fixed in a cradle to minimize motion and the isoflurane level was decreased at 1.5% for anesthesia with a breath mask. The body temperature was maintained at around 37°C. After being placed inside the MRI system, the animals were monitored for the breath rate to be kept at 50–70 per minute by adjusting the isoflurane level at around 1.5%. To examine demyelination effects, 4 rats (2 control and 2 demyelinated rats) with 8 weeks of age were used for the MRI scans. For the 2 rats of demyelination, 0.2% (w/w) cuprizone (bis-cyclohexanone oxaldihydrazone, Sigma-Aldrich Inc., St. Louis, MO, USA) mixed into a ground standard rodent chow from 4 weeks to 8 weeks of age [41 (link)]. In order to study remyelination effects, the cuprizone administration was stopped for the demyelinated rats from 8 weeks of age (the first MRI study) to 9 weeks of age (for one week) and then the second MRI study was performed for the remyelinated rats.