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The Effect of Stereotactic Injections on Demyelination and Remyelination: a Study in the Cuprizone Model

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Abstract

Remyelination is the natural repair mechanism in demyelinating disorders of the central nervous system (CNS) such as multiple sclerosis. Several animal models have been used to study demyelination and remyelination. Among toxic animal models, oral administration of the toxin cuprizone leads to white and gray matter demyelination. In contrast, focal demyelination models include the stereotactic application of a toxin such as lysolecithin or ethidium bromide. The injection procedure generates a local disruption of the blood-brain barrier (BBB) and might thus trigger a local inflammatory reaction and consequently may influence demyelination and remyelination. In order to study such consequences, we applied stereotactic injections in the cuprizone model where demyelination and remyelination are mediated independent of this procedure. Immunohistochemistry was performed to detect the presence of lymphocytes and activated glial cells in the injection area. Blood protein stainings were used to assess the integrity of the BBB and myelin staining to evaluate demyelination and remyelination processes. Stereotactic injection led to a local disruption of the BBB as shown by local extravasation of blood proteins. Along the injection canal, T and B lymphocytes could be detected and there was a tendency of a higher microgliosis and astrocytosis. However, these changes did not influence demyelination and remyelination processes at the site of injection, in the corpus callosum, or in the cerebral cortex. Our results suggest that a local stereotactic injection has no major impact on CNS demyelination and remyelination.

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Abbreviations

BBB:

Blood-brain barrier

CNS:

Central nervous system

EAE:

Experimental autoimmune encephalomyelitis

MS:

Multiple sclerosis

PBS:

Phosphate-buffered saline

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Acknowledgements

We thank I. Cierpka-Leja, S. Lang, and A. Niesel for excellent technical assistance.

Author Contributions

LST, TS, VG, and MS were responsible for the study concept and design. LST and VG performed the experiments and analysis. TW and SG contributed in the evaluation of the results. LST and TW drafted the manuscript. VG, TS, and MS provided the essential revision of the manuscript.

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Authors and Affiliations

  1. Clinical Neuroimmunology and Neurochemistry, Department of Neurology, Hannover Medical School, Carl-Neuberg-Str-1, 30625, Hannover, Germany

    Laura Salinas Tejedor, Tanja Wostradowski, Stefan Gingele, Thomas Skripuletz, Viktoria Gudi & Martin Stangel

  2. Center for Systems Neuroscience, Hannover, Germany

    Laura Salinas Tejedor, Tanja Wostradowski & Martin Stangel

Authors
  1. Laura Salinas Tejedor
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  2. Tanja Wostradowski
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  3. Stefan Gingele
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  4. Thomas Skripuletz
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  5. Viktoria Gudi
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  6. Martin Stangel
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Corresponding author

Correspondence to Viktoria Gudi.

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Conflict of Interest

The authors declare that they have no conflicts of interest.

Funding

The work was partly supported by the German Research Foundation (DFG, FOR 1103, TP1b).

Additional information

Summary Statement

In this study, we compared mice that were fed with cuprizone and underwent a stereotactic injection with mice that received cuprizone only. Despite local disruption of the BBB stereotactic injections, neither changed the course of demyelination or the subsequent remyelination, nor modified glial reactions

Viktoria Gudi and Martin Stangel are equally contributed to this work.

Electronic supplementary material

Suppl. Fig. 1

Experimental design. Mice were fed with cuprizone for 5 weeks to induce demyelination. After week 5 mice were fed with normal chow to allow remyelination. PBS was injected stereotactically in the ventricles of cuprizone treated animals at week 4 which corresponds to the peak of OPC proliferation and microglial activation. Brains were collected and studied at the peak of demyelination (week 5) or during early remyelination (week 5.5). Control animals were treated with cuprizone but did not receive a stereotactic injection. Untreated age-matched control animals were also analyzed. (GIF 36 kb)

High resolution image. (TIFF 292 kb)

Suppl. Fig. 2

Demyelination and remyelination after opening the BBB by stereotactic injection of PBS. Animals treated with cuprizone show loss of myelin protein MOG (a) and MAG (b). Stereotactic injections did not influence endogenous remyelinating mechanisms. Representative brain sections are shown for each group. (GIF 304 kb)

High resolution image. (TIFF 2676 kb)

Suppl. Table 1

Primary antibodies applied in the study. (DOCX 15 kb)

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Tejedor, L.S., Wostradowski, T., Gingele, S. et al. The Effect of Stereotactic Injections on Demyelination and Remyelination: a Study in the Cuprizone Model. J Mol Neurosci 61, 479–488 (2017). https://doi.org/10.1007/s12031-017-0888-y

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  • DOI: https://doi.org/10.1007/s12031-017-0888-y

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