Cuprizone

Manufactured by Merck Group

Sourced in United States, Germany, Macao, China

Cuprizone is a laboratory equipment product manufactured by Merck Group. It is a chemical compound that is primarily used in research applications. The core function of Cuprizone is to induce demyelination in animal models, which is a condition where the protective myelin sheath around nerve fibers is damaged or lost.

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Cuprizone (bis-cyclohexanone oxaldihydrazone) (6 citations) Cuprizone (CPZ) (5 citations) Cuprizone powder (4 citations) Cuprizone diet (1 citations)

132 protocols using cuprizone

Cuprizone-Induced Demyelination Protocol

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Male C57BL/6 mice (8 weeks old) were purchased from Spelford (Beijing) Biotechnology Company. Mice were housed in an air-conditioned room at 22 ± 1^°C with a 12-h light-dark cycle (lights on from 7:00 a.m. to 7:00 p.m.) (Zhao et al., 2021 (link)). Food and tap water were freely available. All procedures for the following experiments were approved by the Animal Care and Use Committee, performed by the NIH Guide for the Care and Use of Laboratory Animals.
Twenty male C57BL/6 mice (8 weeks old) were randomly divided into the control group (n = 10) and the cuprizone (CPZ) group (n = 10), and mice in the CPZ group were fed by a diet containing 0.2% cuprizone (reagent purchased from Sigma, custom-made feed from Jiangsu Hershey Feeds). By referring to the literature (Wellman et al., 2020 (link)), C57BL/6 male mice were selected to be fed 0.2% cuprizone chow to construct the demyelination model, and we chose continuous feeding (11 weeks) for the CPZ group (Zhao et al., 2021 (link)). Then, mice were executed, and the corpus callosum was immediately stored at −80°C for subsequent detection of the targeted lipidomics and transcriptomics.

Zhao Z.J., Zheng R.Z., Wang X.J., Li T.Q., Dong X.H., Zhao C.Y, & Li X.Y. (2022). Integrating Lipidomics and Transcriptomics Reveals the Crosstalk Between Oxidative Stress and Neuroinflammation in Central Nervous System Demyelination. Frontiers in Aging Neuroscience, 14, 870957.

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Induction and Modeling of Experimental Autoimmune Encephalomyelitis

Cited 3 times

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EAE was induced as previously described (57 (link), 71 (link), 72 (link)) using MOG_35–55 peptide (Hooke Laboratories EAE induction kit, EK-2110). Pertussis toxin (200 ng/mouse, i.p.; Hooke Laboratories) was injected on day 0 and 2 relative to immunization. Only animals exhibiting clinical disease were examined by transcriptional analysis. For experiments modeling blood-brain barrier disruption, 200 ng Pertussis toxin was injected i.p. in 500 μL PBS as above at 96 hours and 48 hours prior to adoptive transfer of CD8⁺ T cells. Cuprizone (bis[cyclohexanone]oxaldihydrazone; Sigma-Aldrich) diet containing 0.3% w/w Cuprizone was prepared by Test Diet in 5LG6 base diet and utilized within 6 months of manufacture. Mice were allowed access to experimental or control (5LG6) diet ad libitum and monitored weekly for 6 weeks prior to further manipulation. For intracerebral injections, anesthetized mice were injected with 1.5 μL containing 3 × 10⁹ genomic copies (gc) of AAV1.Syn.OVA.GFP or AAV1.Syn.GFP at 0.5 μL/min.

Clarkson B.D., Grund E.M., Standiford M.M., Mirchia K., Westphal M.S., Muschler L.S, & Howe C.L. (2023). CD8+ T cells recognizing a neuron-restricted antigen injure axons in a model of multiple sclerosis. The Journal of Clinical Investigation, 133(21), e162788.

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Cuprizone-Induced Mouse Demyelination

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For the cuprizone model, adult mice were fed regular chow mixed with 0.3% cuprizone (Sigma, 14,690) ad libitum for 5 weeks to induce demyelination.

Ennerfelt H., Frost E.L., Shapiro D.A., Holliday C., Zengeler K.E., Voithofer G., Bolte A.C., Lammert C.R., Kulas J.A., Ulland T.K, & Lukens J.R. (2022). SYK coordinates neuroprotective microglial responses in neurodegenerative disease. Cell, 185(22), 4135-4152.e22.

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Cuprizone-Induced Demyelination in C57BL/6J Mice

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C57BL/6J male mice (8 weeks old) were obtained from Harlan (UK) and housed (21±2 °C, humidity 36±2% at 12/12-h light/dark cycle) in the Bio-Resource Unit of Dublin City University with food and water provided ad libitum. All procedures were approved by the University Ethics Committee, and licensed by the Department of Children and Health (Rep. of Ireland) in accordance with European Communities Council Directive of 24 November 1986 (86/609/ECC). Special efforts were made to minimize animal suffering and reduce the number of animals used. All animals received a Modified LabDiet®, with 0.2% cuprizone (Sigma, MO) supplemented to chow of the experimental (cuprizone treated) mice for 8 weeks, a time sufficient for induction of demyelination [19] (link).

Bagchi B., Al-Sabi A., Kaza S., Scholz D., O'Leary V.B., Dolly J.O, & Ovsepian S.V. (2014). Disruption of Myelin Leads to Ectopic Expression of KV1.1 Channels with Abnormal Conductivity of Optic Nerve Axons in a Cuprizone-Induced Model of Demyelination. PLoS ONE, 9(2), e87736.

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Cuprizone-induced Chronic Demyelination

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Induction of chronic demyelination disease by Cuprizone was done as previously described.28 (link) Shortly, 5–6-month-old female C57BL/6 mice were feed with 0.3% w/w Cuprizone (biscyclohexane oxaldihydrazone, Sigma-Aldrich, St. Louis, MO) in a diet of ground mouse chow for a period of 10 weeks. Subsequently, mice were sacrificed for western blot analysis (n = 3) or for histological analysis (n = 6). An age-matched control group of normal diet to serve as naïve, nontreated controls (n = 9).

Frid K., Einstein O., Friedman-Levi Y., Binyamin O., Ben-Hur T, & Gabizon R. (2015). Aggregation of MBP in chronic demyelination. Annals of Clinical and Translational Neurology, 2(7), 711-721.

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Cuprizone and EAE Mouse Models

Cited 3 times

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For the cuprizone model, adult male mice (ages in Table S1) were fed regular chow mixed with 0.3% cuprizone (Sigma #14690) ad libitum for 5 weeks to induce demyelination as previously described [55 (link)]. For EAE, Lrp1^fl/flOlig1^Cre and Olig1^Cre mice were immunized and scored as previously described [57 (link)].
Tamoxifen (T5648, Sigma) was prepared at 20mg/mL in 0.2μm filtered corn oil (C8267, Sigma) by shaking overnight at 37°C. Lrp1^fl/flPdgfra^Cre-ERt2 adult mice received two IP injections of 200mg/kg Tamoxifen, two days apart. No more than 4mg Tamoxifen per injection was administered to adult mice. Mice were allowed to recover for 4 weeks after the final Tamoxifen injection before EAE immunizations. Lrp1fl/flPdgfraCre-ERt2− and LRP1fl/flPdgfraCre-ERt2+ mice were immunized twice, three weeks apart, due to the immunomodulatory effects of Tamoxifen [6 (link), 31 (link)]. EAE scores for Lrp1^fl/flPdgfra^Cre-ERt2 mice were collected after the second immunization.

Fernández-Castañeda A., Chappell M.S., Rosen D.A., Seki S.M., Beiter R.M., Johanson D.M., Liskey D., Farber E., Onengut-Gumuscu S., Overall C., Dupree J.L, & Gaultier A. (2019). The active contribution of OPCs to neuroinflammation is mediated by LRP1. Acta neuropathologica, 139(2), 365-382.

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Cuprizone-induced demyelination and progesterone treatment

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Intact 8 week-old C57Bl/6 female mice were used in all experiments, except for one (and in the experiments with transgenic mice) in which mice were ovariectomized one week before the beginning of cuprizone intoxication. Female mice (n =10 per group) were fed for 12 weeks with a powder meal containing 0,2% of the copper chelator cuprizone (Sigma Aldrich, St Louis, MO). Control animals were also fed with the same powder meal, but without cuprizone. After 12 weeks, cuprizone was removed from the diet and mice received for 3 weeks either a 20 mm subcutaneous Silastic implant containing progesterone (Sigma) or an empty implant. Nestorone was administered during 3 weeks by Alzet mini osmotic pumps (model #2004, with a release rate of 0,25µl/h), delivering 1, 4, 6, 8 or 16 µg Nestorone per day, solubilized in 40% molecusol (2-OH-propyl-β-cyclodextrin). This synthetic 19-nor-progesterone derivative is 100 times more active than P on reproductive system end-points (Kumar et al., 2000 (link)). The high potency and selectivity of 19-nor-progesterone derivatives such as Nestorone make them suitable for sustained administration at very low doses via non-oral long-acting delivery systems (Sitruk-Ware and Nath, 2010 (link)).

el-Etr M., Rame M., Boucher C., Ghoumari A., Kumar N., Liere P., Pianos A., Schumacher M, & Sitruk-Ware R. (2014). Progesterone and Nestorone promote myelin regeneration in chronic demyelinating lesions of corpus callosum and cerebral cortex. Glia, 63(1), 104-117.

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Cuprizone-Induced Demyelination Model

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Rats were divided into 3 groups, with 10 rats in each group:

Control group: Rats of this group were given 1.5 mL carboxymethylcellulose (CMC) purchased from Sigma Aldrich (St. Louis, MO, USA) daily by oral gavage for 5 weeks;

Cuprizone (Cup) group: Rats of this group were given 450 mg/kg of Cuprizone purchased from Sigma Aldrich (St. Louis, MO, USA) dissolved in 1.5 mL of 1% CMC per day orally for 5 weeks [5 ];

Cup + L-carnitine (LC) group: Rats of this group were given 450 mg/kg of Cuprizone dissolved in 1.5 mL of 1% CMC, per day orally + 100 mg/kg/day L carnitine purchased from El-Gomhoria Medical Company, Egypt orally for 5 weeks [19 (link)].

Safwat S.M., Aboonq M.S., El Tohamy M., Mojaddidi M., Al-Qahtani S.A., Zakari M.O., ElGendy A.A, & Hussein A.M. (2023). New Insight into the Possible Roles of L-Carnitine in a Rat Model of Multiple Sclerosis. Brain Sciences, 14(1), 23.

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Cuprizone-Induced Mouse Demyelination

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For the cuprizone model, adult mice were fed regular chow mixed with 0.3% cuprizone (Sigma, 14,690) ad libitum for 5 weeks to induce demyelination.

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Cuprizone-induced Demyelination and Recovery

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The cuprizone mouse model was treated according to a modified version of the Armstrong et al. 2002 protocol [18 (link)]. Briefly, 8-week-old C57/BI6 mice were fed with 0.2% (w/w) cuprizone (bis[cyclohexanone]oxaldihydrazone; Sigma-Aldrich) for 5 weeks to induce demyelination, followed by a 9-day recovery period during which cuprizone was removed from the diet and mice were administered with vehicle control or GSK247246 twice daily. GSK247246 was formulated as a suspension using 1% aqueous methylcellulose as the vehicle.

Chen Y., Zhen W., Guo T., Zhao Y., Liu A., Rubio J.P., Krull D., Richardson J.C., Lu H, & Wang R. (2017). Histamine Receptor 3 negatively regulates oligodendrocyte differentiation and remyelination. PLoS ONE, 12(12), e0189380.

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