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Cited by 40Multiple Sclerosis Research StudiesNeurogenesis and neuroplasticity mechanismsNeuroinflammation and Neurodegeneration Mechanisms

Strain differences in cuprizone induced demyelination

Open access
Yu, Qili, Hui, Ryan, Park, Jiyoung, Huang, Yangyang, Kusnecov, Alexander W., Dreyfus, Cheryl F., Zhou, Renping·2017·Cell & Bioscience
DOI 10.1186/s13578-017-0181-3PMC PMC5670722
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Abstract

BACKGROUND: Multiple sclerosis (MS) is a severe neurological disorder, characterized by demyelination of the central nervous system (CNS), and with a prevalence of greater than 2 million people worldwide. In terms of research in MS pathology, the cuprizone toxicity model is widely used. Here we investigated the contribution of genetic differences in response to cuprizone-induced demyelination in two genetically different mouse strains: CD1 and C57BL/6. RESULTS: We demonstrate that exposure to a diet containing 0.2% cuprizone resulted in less severe demyelination in the midline of the corpus callosum over the fornix in CD1 mice than C57BL/6 mice. With continuous cuprizone feeding, demyelination in CD1 mice was not prominent until after 7 weeks, in contrast to C57BL/6 mice, which showed prominent demyelination after 4 weeks of exposure. Concomitantly, immunohistochemical analysis demonstrated more oligodendrocytes, as well as fewer oligodendrocyte progenitor cells, microglia and astrocytes in cuprizone treated CD1 mice. We also analyzed 4-weeks-cuprizone treated corpus callosum tissue samples and found that cuprizone treated CD1 mice showed a smaller reduction of myelin-associated glycoprotein (MAG) and a smaller increase of Iba1 and NG2. CONCLUSIONS: These observations suggest that CD1 mice are less vulnerable to cuprizone-induced demyelination than C57BL/6 mice and thus genetic background factors appear to influence the susceptibility to cuprizone-induced demyelination.

AI snapshot

Written from the paper’s full text.

TL;DR

CD1 mice show less severe and delayed cuprizone-induced demyelination in the corpus callosum than C57BL/6 mice, with a blunted oligodendrocyte loss and reduced OPC, microglial, and astrocyte responses. The authors conclude that genetic background influences susceptibility to cuprizone toxicity.

Key findings

  • With 0.2% cuprizone, C57BL/6 mice showed prominent demyelination after 4 weeks, whereas CD1 mice did not show prominent demyelination until after 7 weeks; CD1 mice had less severe midline corpus callosum demyelination over the fornix.
  • Immunohistochemistry showed more mature GSTpi+ oligodendrocytes and fewer NG2+ OPCs, Iba1+ microglia, and GFAP+ astrocytes in cuprizone-treated CD1 mice than C57BL/6 mice. Significant strain differences were reported for GSTpi at 4, 5, 6, and 7 weeks (P < 0.05 in ANOVA), NG2 at 4 and 6 weeks (P < 0.05), Iba1 at 4, 6, and 7 weeks (P < 0.01), and GFAP at 6 and 7 weeks (P < 0.01).
  • After 4 weeks of cuprizone, western blot showed a smaller MAG reduction in CD1 mice than C57BL/6 mice (26% reduction in CD1 vs 73% in C57BL/6; P < 0.05), and smaller increases in Iba1 (P < 0.01) and NG2 (P < 0.05). MBP reduction and GFAP induction did not reach statistical significance between strains.
  • CD1 and C57BL/6 mice consumed similar amounts of cuprizone diet per body weight, despite CD1 mice being heavier and eating more diet.

Methods

  • CD1 mice (Charles River, strain code 022) and C57BL/6J mice (Jackson Laboratory, stock No. 000664) were purchased at 8 weeks of age, acclimated 1 week, and fed a 0.2% cuprizone-containing diet or control diet ad libitum for 4 to 7 weeks.
  • Histology used Black gold staining and Luxol fast blue–periodic acid Schiff (LFB–PAS) staining, with blind scoring on a 0–3 scale. Immunohistochemistry used GSTpi for mature oligodendrocytes, NG2 for OPCs, Iba1 for microglia/macrophages, and GFAP for astrocytes, with confocal imaging of the midline corpus callosum above the fornix.
  • Western blot analysis of 4-week cuprizone-treated corpus callosum measured MBP, MAG, Iba1, GFAP, and NG2, normalized to β-tubulin. Statistical comparisons used unpaired Student’s t-test or two-way ANOVA where appropriate, with significance at P < 0.05 and data presented as mean ± SEM.

Limitations

  • The paper does not include a formal limitations section. It states that the exact mechanism of cuprizone-induced demyelination remains elusive and that future experiments addressing pro-inflammatory cytokines and functional interference with microglia and/or astrocytes are needed.
  • Evident from the text: the study compares only two mouse strains, uses a single cuprizone dose (0.2%), and focuses on the midline corpus callosum over the fornix, so generalizability to other strains, doses, and CNS regions is limited.
  • Several reported differences were not statistically significant, including MBP reduction, GFAP induction by western blot, and the 7-week LFB–PAS strain difference; the authors also note a lateral corpus callosum demyelination pattern in about 10% of cuprizone-treated CD1 mice but do not show those data.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s13578-017-0181-3

Cites

38 of 38 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

17 of 42 cited sources checked · 8 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    Cuprizone selectively affects oligodendrocytes.

    “Nevertheless, a consensus seems to be that cuprizone selectively affects oligodendrocytes.”
    Quote in the paperSubstantial claim1 of 10 sources checked
    • cites [6] Hiremath MM, Saito Y, Knapp GW, Ting JP, Suzuki K, Matsushima GK. Microglial/macrophage accumulation during cuprizone-induced demyelination in C57BL/6 mice. J Neuroimmunol. 1998;92:38–49. source
    • cites [8] Ludwin SK. Central nervous system demyelination and remyelination in the mouse: an ultrastructural study of cuprizone toxicity. Lab Invest. 1978;39:597–612.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [19] Tandler B, Hoppel CL. Division of giant mitochondria during recovery from cuprizone intoxication. J Cell Biol. 1973;56:266–72. source

      The Introduction and Results (pp. 266–267) describe cuprizone-induced changes in mouse hepatocytes, and the Summary reports greatly enlarged hepatic mitochondria in cuprizone-fed mice. The paper does not discuss oligodendrocytes; its finding of an effect in liver cells runs contrary to the unqualified claim that cuprizone selectively affects oligodendrocytes. (LLM, conf=0.84)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [20] Asano M, Wakabayashi T, Ishikawa K, Kishimoto H. Mechanism of the formation of megamitochondria by copper-chelating agents. IV. Role of fusion phenomenon in the cuprizone-induced megamitochondrial formation. Acta Pathol Jpn. 1978;28:205–13. source
    • cites [21] Wakabayashi T, Asano M, Ishikawa K, Kishimoto H. Mechanism of the formation of megamitochondria by copper-chelating agents. V. Further studies on isolated megamitochondria. Acta Pathol Jpn. 1978;28:215–23. source
    • cites [22] Biancotti JC, Kumar S, de Vellis J. Activation of inflammatory response by a combination of growth factors in cuprizone-induced demyelinated brain leads to myelin repair. Neurochem Res. 2008;33:2615–28. source
    • cites [23] Acs P, Komoly S. Selective ultrastructural vulnerability in the cuprizone-induced experimental demyelination. Ideggyogy Sz. 2012;65:266–70.
    • cites [18] Praet J, Guglielmetti C, Berneman Z, Van der Linden A, Ponsaerts P. Cellular and molecular neuropathology of the cuprizone mouse model: clinical relevance for multiple sclerosis. Neurosci Biobehav Rev. 2014;47:485–505. source
    • cites [24] Acs P, Selak MA, Komoly S, Kalman B. Distribution of oligodendrocyte loss and mitochondrial toxicity in the cuprizone-induced experimental demyelination model. J Neuroimmunol. 2013;262:128–31. source
    • cites [25] Tandler B, Hoppel CL. The failure of supplemental dietary copper to prevent cuprizone-induced alterations in mouse hepatocytes. Beitr Pathol. 1975;156:56–64. source
  • Findingown data

    The C57BL/6 mice showed a dramatic loss of myelin staining at 4 weeks of cuprizone treatment.

    “The C57BL/6 mice showed a dramatic loss of myelin staining at this time point (Fig. 1b), which is consistent with previous reports [5].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [5] Matsushima GK, Morell P. The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol. 2001;11:107–16. source

      The opening passage (section and page unavailable) describes massive cuprizone-induced demyelination in mouse brain and specifically discusses C57BL/6 mice fed cuprizone. It does not establish that a dramatic loss of myelin staining occurs at four weeks, so the cited report supports the general finding but not that time point or staining observation. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
  • Interpretationreasoning

    The cuprizone model is particularly suitable for studying subtypes of MS which show primary oligodendrocyte dystrophy.

    “This actually makes the cuprizone model particularly suitable for studying subtypes of MS which show primary oligodendrocyte dystrophy, reminiscent of toxin- or virus-, rather than autoimmunity, induced demyelination [31, 32].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [31] Lucchinetti C, Bruck W, Parisi J, Scheithauer B, Rodriguez M, Lassmann H. Heterogeneity of multiple sclerosis lesions: implications for the pathogenesis of demyelination. Ann Neurol. 2000;47:707–17. source
    • cites [32] Stys PK, Zamponi GW, van Minnen J, Geurts JJ. Will the real multiple sclerosis please stand up? Nat Rev Neurosci. 2012;13:507–14. source

      The Conclusions and future directions section says models of primary damage to oligodendrocytes, myelin and axons may be more relevant and informative for MS, and identifies cuprizone as one such model. The Potential mechanisms section reports oligodendroglial damage and demyelination with little inflammation. The paper presents this relevance as a possibility, however; it does not establish specific MS subtypes with primary oligodendrocyte dystrophy or make the sentence’s virus-induced comparison. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
  • Interpretationreasoning

    A relatively abundant MAG level might indicate a relatively intact myelin-axon interface in the CD1 mice, albeit a damaged outer myelin sheath.

    “a relatively abundant MAG level might indicate a relatively intact myelin-axon interface in the CD1 mice, albeit a damaged outer myelin sheath.”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [35] Trapp BD, Quarles RH. Immunocytochemical localization of the myelin-associated glycoprotein. Fact or artifact? J Neuroimmunol. 1984;6:231–49. source
    • cites [36] Trapp BD. Myelin-associated glycoprotein. Location and potential functions. Ann NY Acad Sci. 1990;605:29–43. source
    • cites [37] Laule C, Vavasour IM, Kolind SH, Li DKB, Traboulsee TL, Moore GRW, MacKay AL. Magnetic resonance imaging of myelin. Neurotherapeutics. 2007;4:460–84. source

      Section 4 describes a possible role for MAG in interactions between oligodendrocytes and axons, which lends limited support to the proposed interpretation. It does not report MAG levels in CD1 mice or establish that abundant MAG indicates an intact myelin–axon interface despite damage to the outer sheath. No page number is available in our copy. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Premisecitation

    Demyelination in the cortex induced by cuprizone treatment has also been shown by Skripuletz et al. to be affected by genetic background factors.

    “In addition to the corpus callosum, demyelination in the cortex induced by cuprizone treatment has also been shown by Skripuletz et al. to be affected by genetic background factors [9].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [9] Skripuletz T, Lindner M, Kotsiari A, Garde N, Fokuhl J, Linsmeier F, Trebst C, Stangel M. Cortical demyelination is prominent in the murine cuprizone model and is strain-dependent. Am J Pathol. 2008;172:1053–61. source
  • Classified as probably not a substantial claim. 13
  • May combine two claims. 16
  • The quote only partly states this claim. 2

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Other mouse strains are also used sometimes in the cuprizone model.

    “However, other strains are also used sometimes [7–10].”
    Quote in the paperNot a substantial claim2 of 4 sources checked
    • cites [7] Blakemore WF. Observations on oligodendrocyte degeneration, the resolution of status spongiosus and remyelination in cuprizone intoxication in mice. J Neurocytol. 1972;1:413–26. source
    • cites [8] Ludwin SK. Central nervous system demyelination and remyelination in the mouse: an ultrastructural study of cuprizone toxicity. Lab Invest. 1978;39:597–612.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [9] Skripuletz T, Lindner M, Kotsiari A, Garde N, Fokuhl J, Linsmeier F, Trebst C, Stangel M. Cortical demyelination is prominent in the murine cuprizone model and is strain-dependent. Am J Pathol. 2008;172:1053–61. source
    • cites [10] Taylor LC, Gilmore W, Matsushima GK. SJL mice exposed to cuprizone intoxication reveal strain and gender pattern differences in demyelination. Brain Pathol. 2009;19:467–79. source

  • Classified as probably not a substantial claim.

    Premisecitation

    The cuprizone model involves administration of cuprizone to induce oligodendrocyte apoptosis.

    “The cuprizone model involves administration of the toxin cuprizone to induce oligodendrocyte apoptosis [4, 5].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [4] Torkildsen O, Brunborg LA, Myhr KM, Bo L. The cuprizone model for demyelination. Acta Neurol Scand Suppl. 2008;188:72–6. source

      The “Mice strain and cuprizone administration” section says mice are fed cuprizone, leading to oligodendrocyte death. “Effects of cuprizone administration” describes apoptosis as a plausible hypothesis for that death, rather than establishing it as the mechanism. The claim states apoptosis without that qualification. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [5] Matsushima GK, Morell P. The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol. 2001;11:107–16. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Upon treatment with cuprizone, mice typically exhibit profound demyelination in the corpus callosum, the cortex and the superior cerebellar peduncles.

    “Upon treatment with cuprizone, mice typically exhibit profound demyelination in the corpus callosum, the cortex and the superior cerebellar peduncles [4, 5].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [4] Torkildsen O, Brunborg LA, Myhr KM, Bo L. The cuprizone model for demyelination. Acta Neurol Scand Suppl. 2008;188:72–6. source
    • cites [5] Matsushima GK, Morell P. The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol. 2001;11:107–16. source

      The opening passage in our copy (section and page not labeled) says cuprizone reproducibly induces massive demyelination in large areas of mouse brain. The references include a study titled “Demyelination of the superior cerebellar peduncle in the mouse induced by cuprizone,” but no passage in our copy establishes that profound demyelination typically occurs in all three named regions; the corpus callosum and cortex are not identified. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In the corpus callosum, cuprizone treatment induces apoptosis of oligodendrocytes together with recruitment of microglia/macrophages, astrocytes and OPCs.

    “In the corpus callosum, apoptosis of oligodendrocytes is observed, together with recruitment of microglia/macrophages, astrocytes and oligodendrocyte progenitor cells (OPCs) [5, 6].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [5] Matsushima GK, Morell P. The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol. 2001;11:107–16. source

      The main-text passage supplied in our copy (no page label) says that cuprizone-fed mice undergo mature oligodendrocyte apoptosis, followed by microglial recruitment, and that oligodendrocyte progenitors proliferate and invade demyelinated areas. It does not specify the corpus callosum or say that astrocytes are recruited; the reference-list mentions of astrocytes do not establish that part of the claim. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [6] Hiremath MM, Saito Y, Knapp GW, Ting JP, Suzuki K, Matsushima GK. Microglial/macrophage accumulation during cuprizone-induced demyelination in C57BL/6 mice. J Neuroimmunol. 1998;92:38–49. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Most investigators use the standard C57BL/6 mouse strain in the cuprizone model to study molecular components of corpus callosum demyelination.

    “Most investigators use the standard C57BL/6 mouse strain in the cuprizone model to study the different molecular components involved in the complex pathogenic processes leading to corpus callosum demyelination [5].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [5] Matsushima GK, Morell P. The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol. 2001;11:107–16. source

      The opening summary (page not supplied) describes cuprizone-induced demyelination in C57BL/6 mice and studies of myelin gene expression and cellular events. It does not establish that most investigators use this strain or specify the corpus callosum as the site studied. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    SJL mice have been shown to be less susceptible to demyelination in the corpus callosum compared to the established C57BL/6 strain.

    “For instance, SJL mice have been shown to be less susceptible to demyelination in the corpus callosum compared to the established C57BL/6 strain [10].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [10] Taylor LC, Gilmore W, Matsushima GK. SJL mice exposed to cuprizone intoxication reveal strain and gender pattern differences in demyelination. Brain Pathol. 2009;19:467–79. source

  • May combine two claims.

    Premisecitation

    In their study the authors challenged BALB/cJ mice with 0.2% cuprizone treatment of 6 weeks, and observed that cortical demyelination of BALB/cJ mice was incomplete, as opposed to C57BL/6 strain which shows no detectable cortical myelin at this stage.

    “In their study the authors challenged BALB/cJ mice with 0.2% cuprizone treatment of 6 weeks, and observed that cortical demyelination of BALB/cJ mice was incomplete, as opposed to C57BL/6 strain which shows no detectable cortical myelin at this stage.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [9] Skripuletz T, Lindner M, Kotsiari A, Garde N, Fokuhl J, Linsmeier F, Trebst C, Stangel M. Cortical demyelination is prominent in the murine cuprizone model and is strain-dependent. Am J Pathol. 2008;172:1053–61. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    SJL mice exhibit a unique pattern of demyelination in that the area of the corpus callosum that is immediately lateral to midline seems to demyelinate more prominently than the midline.

    “The authors also reported that SJL mice exhibit a unique pattern of demyelination in that the area of the corpus callosum that is immediately lateral to midline seems to demyelinate more prominently than the midline.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [10] Taylor LC, Gilmore W, Matsushima GK. SJL mice exposed to cuprizone intoxication reveal strain and gender pattern differences in demyelination. Brain Pathol. 2009;19:467–79. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A switch from Caspase 3-dependent to Caspase 3-independent form of apoptosis is reported for later stages of cuprizone exposure.

    “(1), a switch from Caspase 3-dependent to Caspase 3-independent form of apoptosis is reported for later stages of cuprizone exposure [33];”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Hesse A, Wagner M, Held J, Bruck W, Salinas-Riester G, Hao Z, Waisman A, Kuhlmann T. In toxic demyelination oligodendroglial cell death occurs early and is FAS independent. Neurobiol Dis. 2010;37:362–9. source

      The Results report that activated caspase 3 was present in 97% of apoptotic cells on day 6 but absent from dying cells on day 21. The Discussion describes this as suggestive of a switch to caspase 3-independent cell death; it only speculates that apoptosis-like mechanisms contribute at later stages, rather than establishing a caspase 3-independent form of apoptosis. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    On average, CD1 mice showed a milder reduction in MBP protein levels after 4 weeks of cuprizone treatment compared to C57BL/6 mice, but the difference did not reach statistical significance.

    “As shown in Fig. 4a, c, on average, CD1 mice showed a milder reduction in MBP protein levels after cuprizone treatment compared to C57BL/6 mice, however the difference did not reach statistical significance (P > 0.05 in t-test, N = 6 for C57BL/6 and N = 8 for CD1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    With continuous cuprizone feeding, demyelination in CD1 mice was not prominent until after 7 weeks, whereas C57BL/6 mice showed prominent demyelination after 4 weeks of exposure.

    “With continuous cuprizone feeding, demyelination in CD1 mice was not prominent until after 7 weeks, in contrast to C57BL/6 mice, which showed prominent demyelination after 4 weeks of exposure.”
    • Also stated in: § Results · plain
      “The C57BL/6 mice showed a dramatic loss of myelin staining at this time point (Fig. 1b), which is consistent with previous reports [5]. However, the CD1 mice didn’t show significant myelin loss.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In C57BL/6 mice, NG2+ OPC number peaked at 4 and 6 weeks of cuprizone treatment, while CD1 mice exhibited a gradual increase in NG2+ cell population.

    “In C57BL/6 mice, NG2+ OPC number peaked at 4 and 6 weeks, while CD1 mice exhibited a gradual increase in NG2+ cell population, coinciding with increasingly severe demyelination.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    By 7 weeks of cuprizone treatment, CD1 mice were slightly less demyelinated than C57BL/6 mice, but this difference was not statistically significant.

    “By 7 weeks, CD1 mice were slightly less demyelinated compared to C57BL/6 mice, but this difference is not statistically significant, although the Black gold data shows CD1 mice still exhibit more Black gold-stained fibers at this point.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Cuprizone-treated CD1 mice have fewer astrocytes than cuprizone-treated C57BL/6 mice.

    “Concomitantly, immunohistochemical analysis demonstrated more oligodendrocytes, as well as fewer oligodendrocyte progenitor cells, microglia and astrocytes in cuprizone treated CD1 mice.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Cuprizone-treated CD1 mice have fewer oligodendrocyte progenitor cells than cuprizone-treated C57BL/6 mice.

    “Concomitantly, immunohistochemical analysis demonstrated more oligodendrocytes, as well as fewer oligodendrocyte progenitor cells, microglia and astrocytes in cuprizone treated CD1 mice.”
    • Also stated in: § Results · plain
      “Although CD1 mice showed a much smaller increase in NG2+ cells at 4 and 6 weeks (P < 0.05 in ANOVA for both 4 and 6 weeks), they do show comparable numbers of NG2+ cells as C57BL/6 strain at 7 weeks.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD1 mice showed a much smaller increase in NG2+ cells at 4 and 6 weeks of cuprizone treatment than C57BL/6 mice, but comparable numbers at 7 weeks.

    “Although CD1 mice showed a much smaller increase in NG2+ cells at 4 and 6 weeks (P < 0.05 in ANOVA for both 4 and 6 weeks), they do show comparable numbers of NG2+ cells as C57BL/6 strain at 7 weeks.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Immunohistochemistry data show that CD1 mice exhibit less oligodendrocyte decrease and concomitantly less recruitment of OPCs/microglia/astrocytes following cuprizone intoxication.

    “Second, immunohistochemistry data show that CD1 mice exhibit less oligodendrocyte decrease and concomitantly less recruitment of OPCs/microglia/astrocytes following cuprizone intoxication.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Following 4 weeks of cuprizone treatment, CD1 mice exhibited on average a smaller induction of GFAP compared to C57BL/6 mice, but the difference did not reach statistical significance.

    “Western blot analysis examining GFAP levels in the two strains following 4 weeks of cuprizone treatment showed that CD1 mice exhibited on average a smaller induction of GFAP, which is consistent with the immunohistochemical analysis, but the difference did not reach statistical significance (Fig. 4a, f).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Compared to C57BL/6 mice, CD1 mice showed on average a lower degree of astrocyte recruitment at all time points, with significant difference at 6 and 7 weeks of cuprizone treatment.

    “Compared to C57BL/6 mice, CD1 mice showed on average, a lower degree of astrocyte recruitment at all time points and statistical comparison between strains showed significant difference at 6 and 7 weeks of cuprizone treatment (Fig. 7).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Both strains exhibited a gradual increase in the GFAP and to-pro-3 double positive astrocyte population from 4 to 7 weeks of cuprizone treatment, with 7 weeks showing the greatest amount of astrogliosis.

    “Both strains exhibited a gradual increase in the GFAP and to-pro-3 double positive astrocyte population from 4 to 7 weeks of treatment, with 7 weeks showing the greatest amount of astrogliosis.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The level of Iba1 and to-pro-3 double positive microglia peaked at 4 weeks of cuprizone exposure in C57BL/6 mice, and then gradually decreased in number at 5-7 weeks.

    “the level of Iba1 and to-pro-3 double positive microglia peaked at 4 weeks of cuprizone exposure in C57BL/6 mice, and then gradually decreased in number at 5–7 weeks.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    For all cell counting, positively stained cells were identified by antibody-to-pro-3 colocalization.

    “For all cell counting, positively stained cells are identified by antibody-to-pro-3 colocalization (Additional file 1: Figure S2).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The cuprizone neurotoxicity animal model is widely used in MS research.

    “The cuprizone neurotoxicity animal model is widely used in MS research [3].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [3] Kipp M, Clarner T, Dang J, Copray S, Beyer C. The cuprizone animal model: new insights into an old story. Acta Neuropathol. 2009;118:723–36. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Sections for LFB-PAS analysis were scored blind on a scale of 0-3.

    “For analysis, the sections were scored blind on a scale of 0–3 (Additional file 1: Figure S1) by judging the relative intensity of blue (myelin content) and pink (demyelinated area).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Study design and conductnone given

    For histology and immunohistochemistry, at least 3 sections were stained per mouse and the average myelin scoring or cell count/mm2 was taken to represent a single mouse.

    “For any of the subsequent analyses, including histology and immunohistochemistry, at least 3 sections were stained per mouse and the average luxol fast blue myelin scoring or cell count/mm2 was taken to represent a single mouse.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Cuprizone-induced apoptosis precedes massive demyelination, indicating that oligodendrocyte degeneration is the main cause, as opposed to a secondary immune response.

    “Importantly this apoptosis precedes the massive demyelination caused by cuprizone administration [18], indicating that oligodendrocyte degeneration is the main cause, as opposed to a secondary immune response.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [18] Praet J, Guglielmetti C, Berneman Z, Van der Linden A, Ponsaerts P. Cellular and molecular neuropathology of the cuprizone mouse model: clinical relevance for multiple sclerosis. Neurosci Biobehav Rev. 2014;47:485–505. source

  • May combine two claims.

    Premisecitation

    The cuprizone induced demyelination in the corpus callosum is best documented in the commonly used C57BL/6 strain and at a dosage of 0.2%.

    “However, the cuprizone induced demyelination in the corpus callosum is best documented in the commonly used C57BL/6 strain and at a dosage of 0.2% [6, 15].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [6] Hiremath MM, Saito Y, Knapp GW, Ting JP, Suzuki K, Matsushima GK. Microglial/macrophage accumulation during cuprizone-induced demyelination in C57BL/6 mice. J Neuroimmunol. 1998;92:38–49. source
    • cites [15] Mason JL, Jones JJ, Taniike M, Morell P, Suzuki K, Matsushima GK. Mature oligodendrocyte apoptosis precedes IGF-1 production and oligodendrocyte progenitor accumulation and differentiation during demyelination/remyelination. J Neurosci Res. 2000;61:251–62. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Microglial processes were observed in between myelin lamellae, possibly stripping myelin from axons.

    “(4), microglial processes were observed in between myelin lamellae, possibly stripping myelin from axons [8].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [8] Ludwin SK. Central nervous system demyelination and remyelination in the mouse: an ultrastructural study of cuprizone toxicity. Lab Invest. 1978;39:597–612.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data

    CD1 mice appear to show relatively mild demyelination and a blunted immune response compared to C57BL/6 mice after the same dosage of cuprizone treatment.

    “CD1 mice appear to show relatively mild demyelination and a blunted immune response compared to C57BL/6 mice after the same dosage of cuprizone treatment.”
    Quote in the paperSubstantial claim
29 of 29

  • The paper relies on this without support.

    CD1 and C57BL/6 mice differ in genetic background.

    “Here we investigated the contribution of genetic differences in response to cuprizone-induced demyelination in two genetically different mouse strains: CD1 and C57BL/6.”

  • The paper relies on this without support.

    Cuprizone intake or exposure was comparable between CD1 and C57BL/6 mice.

    “For analysis of cuprizone-containing diet intake, both body weights (recorded three times weekly) and food consumption (recorded daily) were closely monitored.”

  • The paper relies on this without support.

    The cuprizone-induced demyelination model is relevant to the pathological process of human multiple sclerosis.

    “The cuprizone model of induced demyelination has been widely adopted to study the pathological process of MS for the past 20 years.”

  • The paper relies on this without support.

    Differences between CD1 and C57BL/6 mice in cuprizone-induced demyelination are attributable to genetic background differences.

    “we addressed the important role of genetic background in affecting the demyelination response induced by the neurotoxin cuprizone by comparing CD1 strain versus the commonly used C57BL/6 strain”
4 of 4

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Limitations of the CPZ model include the absence of autoimmune components and variability in susceptibility due to animal- and protocol-related factors.

    “Limitations of this model still include the absence of autoimmune components and variability in susceptibility due to animal- and protocol-related factors137.”

    From From insights to innovations: evaluating preclinical paradigms in demyelinating disease therapeutics (2026) · cites this paper as [137]

    The Discussion (page unavailable) describes cuprizone demyelination as driven primarily by oligodendrocyte injury rather than autoimmunity, and the Conclusions (page unavailable) identify mouse strain as a factor in susceptibility. The paper discusses treatment dose and duration, but does not establish that protocol-related factors cause variability in susceptibility; it also does not explicitly establish a complete absence of autoimmune components. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

Cites

Flagged first, then by citations

1 of the 39 references couldn’t be matched to a known paper.

Research works printed but not linked: 1

  1. Franklin KBJ. The mouse brain in stereotaxic coordinates/Keith B.J. Franklin, George Paxinos. San Diego: Academic Press; 1997.

Cited by

Sorted by citations

Related

Authors
Yu, Q., Hui, R., Park, J., Huang, Y., Kusnecov, A. W., Dreyfus, C. F., & Zhou, R.
Affiliations
Rutgers, The State University of New Jersey (US)
Corresponding author
Zhou, Renping
ORCID
Venue
Cell & Bioscience
Publisher
Springer Science and Business Media LLC
Published
2017-11-03
Volume
7
Issue
1
Pages
59-59
ISSN
2045-3701 (ISSN-L)
DOI
10.1186/s13578-017-0181-3
PMC
PMC5670722
OpenAlex
W2766251897
Cited by
40
References
39
Citations by year
2026: 8 · 2025: 2 · 2024: 4 · 2023: 4 · 2022: 5 · 2021: 2 · 2020: 3 · 2019: 6 · 2018: 5 · 2017: 1
FWCI
2.80
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell & Bioscience (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • National Institutes of Health (2PO1HD023315, RO1EY019012, NS036647, MH104800, MH108994)
  • National Multiple Sclerosis Society (RG4257B4/1)
  • Rutgers (P30ES005022)
Topics
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Multiple Sclerosis Research Studies
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
cuprizone, demyelination, multiple sclerosis, genetic strain differences, ICR mice, C57BL/6 mice, corpus callosum, oligodendrocytes, oligodendrocyte precursor cells, microglia, astrocytes, myelin-associated glycoprotein, Iba1, NG2

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-13.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-13.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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