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Direct, long-term intrathecal application of therapeutics to the rodent CNS

Nature Protocols volume 12, pages 104–121 (2017)Cite this article

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Abstract

Systemic application of therapeutics to the CNS tissue often results in subtherapeutic drug levels, because of restricted and selective penetration through the blood–brain barrier (BBB). Here, we give a detailed description of a standardized technique for intrathecal drug delivery in rodents, analogous to the technique used in humans. The intrathecal drug delivery method bypasses the BBB and thereby offers key advantages over oral or intravenous administration, such as maximized local drug doses with minimal systemic side effects. We describe how to deliver antibodies or drugs over several days or weeks from a s.c. minipump and a fine catheter inserted into the subdural space over the spinal cord (20 min operative time) or into the cisterna magna (10 min operative time). Drug levels can be sampled by quick and minimally invasive cerebrospinal fluid (CSF) collection from the cisterna magna (5 min procedure time). These techniques enable targeted application of any compound to the CNS for therapeutic studies in a wide range of CNS disease rodent models. Basic surgery skills are helpful for carrying out the procedures described in this protocol.

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Figure 1: Histological appearance of neuronal architecture, myelin or astrocyte and microglia reactivity 2 h and 5 d after catheter insertion over the lumbar spinal cord.
Figure 2: Assembly of catheters.
Figure 3: Catheter implantation into the intrathecal space of the lumbar spinal cord—surgery protocol.
Figure 4: Catheter implantation into the cisterna magna—surgery protocol.
Figure 5: Forelimb and hind-limb fine motor skill performance of rats after intrathecal catheter implantation.

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Acknowledgements

This study was supported by grants from the Swiss National Science Foundation (to M.E.S.), the Christopher and Dana Reeve Foundation, the Swiss MS Society, the Hartmann-Müller Foundation, Zurich, the Desirée-and-Niels-Yde Foundation (to B.V.I.) and two MD–PhD fellowships from the Swiss National Science Foundation (no. 323530_151488, to B.V.I., and no. 323630_151489, to M.P.S.). We thank N.K. Medtner and F. Busoni for help with the surgery.

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Authors and Affiliations

  1. Brain Research Institute, University of Zurich, Zurich, Switzerland

    Benjamin V Ineichen, Lisa Schnell, Miriam Gullo, Julia Kaiser, Marc P Schneider, Alice C Mosberger, Nicolas Good & Martin E Schwab

  2. Department of Health Sciences and Technology, ETH Zurich, Zurich, Switzerland

    Benjamin V Ineichen, Lisa Schnell, Miriam Gullo, Julia Kaiser, Marc P Schneider, Alice C Mosberger, Nicolas Good & Martin E Schwab

  3. Clinic of Neurology, Helios Klinik Hagen-Ambrock, Hagen, Germany

    Michael Linnebank

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Contributions

B.V.I., L.S. and M.G. developed the operations. B.V.I., J.K. and M.P.S. performed the operations. J.K., B.V.I., N.G. and A.C.M. provided figures. J.K. designed graphics and processed videos. B.V.I., M.E.S. and M.P.S. wrote the manuscript. M.E.S., B.V.I. and M.L. contributed to conceptual development.

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Correspondence to Benjamin V Ineichen.

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Supplementary information

Supplementary Figures 1 and 2

Supplementary Figure 1 Direct intrathecal application of antibodies and drugs to the rodent central nervous system: spinal cord. Supplementary Figure 2 Direct intrathecal application of antibodies and drugs to the rodent central nervous system: cisterna magna. (PDF 1747 kb)

Catheter implantation over the lumbar spinal cord. (MP4 28825 kb)

Catheter implantation into the cisterna magna. (MOV 30308 kb)

Cerebrospinal fluid sampling. (MP4 7239 kb)

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Ineichen, B., Schnell, L., Gullo, M. et al. Direct, long-term intrathecal application of therapeutics to the rodent CNS. Nat Protoc 12, 104–121 (2017). https://doi.org/10.1038/nprot.2016.151

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