Advertisement

Detection of Neurocan in Cerebrospinal Fluid

  • Uwe RauchEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 836)

Abstract

Cerebrospinal fluid (CFS) is the most easily accessible component of the human central nervous system and has been successfully used for the analysis of disease-associated molecular imbalances, particularly for extracellular matrix components. Alterations in the presence of the nervous system-associated chondroitin sulfate proteoglycan neurocan had been reported from active multiple sclerosis lesions. Neurocan could be detected as a component of human CFS after enrichment of proteoglycans by anion exchange chromatography from pooled liquor as well as individual 300 μL samples by Western blot. However, a general alteration in neurocan levels in CFS sample with high immunoglobulin content could not be demonstrated. To further reduce the sample size, the development of a PG capturing assay based on polybrene-coated 96-well plates was initiated. This approach could be an interesting alternative option for the analysis of PGs in biological fluid and tissue samples.

Key words

Neurocan Chondroitin sulfate proteoglycan Cerebrospinal fluid Anion exchange chromatography Polybrene 

Notes

Acknowledgments

I would like to thank Dr. A. Grubb and J. Warenholt for help with the human CSF samples, and the Alfred Österlunds, the H and J Forssmans, the G and J Kocks, and the Crafoords foundations, the Swedish Research Council and Lunds Universities Medical Faculty for support.

References

  1. 1.
    Rauch, U., Feng, K., and Zhou, X. H. (2001) Neurocan: a brain chondroitin sulfate proteoglycan. Cell. Mol. Life Sci. 58, 1842–1856.PubMedCrossRefGoogle Scholar
  2. 2.
    Rauch, U. (2004) Extracellular matrix components associated with remodeling processes in brain. Cell. Mol. Life Sci. 61, 2031–2045.PubMedCrossRefGoogle Scholar
  3. 3.
    Rauch, U., Gao, P., Janetzko, A., Flaccus, A., Hilgenberg, L., Tekotte, H., et al. (1991) Isolation and characterization of developmentally regulated chondroitin sulfate and chondroitin/keratan sulfate proteoglycans of brain identified with monoclonal antibodies. J. Biol. Chem. 266, 14785–14801.PubMedGoogle Scholar
  4. 4.
    Sobel, R. A. and Ahmed, A. S. (2001) White matter extracellular matrix chondroitin sulfate/dermatan sulfate proteoglycans in multiple sclerosis. J. Neuropathol. Exp. Neurol. 60, 1198–1207.PubMedGoogle Scholar
  5. 5.
    Buss, A., Pech, K., Kakulas, B. A., Martin, D., Schoenen, J., Noth, J., et al. (2009) NG2 and phosphacan are present in the astroglial scar after human traumatic spinal cord injury. BMC Neurol. 9, 32.PubMedCrossRefGoogle Scholar
  6. 6.
    McKeon, R. J., Jurynec, M. J., and Buck, C. R. (1999) The chondroitin sulfate proteoglycans neurocan and phosphacan are expressed by reactive astrocytes in the chronic CNS glial scar. J. Neurosci. 19, 10778–10788.PubMedGoogle Scholar
  7. 7.
    Asher, R. A., Morgenstern, D. A., Fidler, P. S., Adcock, K. H., Oohira, A., Braistead, J. E., et al. (2000) Neurocan is upregulated in injured brain and in cytokine-treated astrocytes. J. Neurosci. 20, 2427–2438.PubMedGoogle Scholar
  8. 8.
    Huang, X., Kim, J. M., Kong, T. H., Park, S. R., Ha, Y., Kim, M. H., et al. (2009) GM-CSF inhibits glial scar formation and shows long-term protective effect after spinal cord injury. J. Neurol. Sci. 277, 87–97.PubMedCrossRefGoogle Scholar
  9. 9.
    Leppert, D., Ford, J., Stabler, G., Grygar, C., Lienert, C., Huber, S., et al. (1998) Matrix metalloproteinase-9 (gelatinase B) is selectively elevated in CSF during relapses and stable phases of multiple sclerosis. Brain 121 (Pt 12), 2327–2334.PubMedCrossRefGoogle Scholar
  10. 10.
    Saez-Valero, J., Costell, M., Sjogren, M., Andreasen, N., Blennow, K., and Luque, J. M. (2003) Altered levels of cerebrospinal fluid reelin in frontotemporal dementia and Alzheimer’s disease. J. Neurosci. Res. 72, 132–136.PubMedCrossRefGoogle Scholar
  11. 11.
    Rauch, U., Hirakawa, S., Oohashi, T., Kappler, J., and Roos, G. (2004) Cartilage link protein interacts with neurocan, which shows hyaluronan binding characteristics different from CD44 and TSG-6. Matrix Biol. 22, 629–639.PubMedCrossRefGoogle Scholar
  12. 12.
    Feng, K., Arnold-Ammer, I., and Rauch, U. (2000) Neurocan is a heparin binding proteoglycan. Biochem. Biophys. Res. Commun. 272, 449–455.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Vascular Wall Biology, Institute of Experimental Medical SciencesLunds UniversityLundSweden

Personalised recommendations