Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration

Brit Mollenhauer, Valerie Cullen, Ilana Kahn, Bryan Krastins, Tiago F. Outeiro, Imelda Pepivani, Juliana Ng, Walter Schulz-Schaeffer, Hans A. Kretzschmar, Pamela J. McLean, Claudia Trenkwalder, David A. Sarracino, Jean Paul VonSattel, Joseph J. Locascio, Omar Ali El-Agnaf, Michael G. Schlossmacher

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Abstract

Because accumulation of α-synuclein (αS) in the brain is a hallmark of Parkinson disease (PD) and related disorders, we examined its occurrence in human cerebrospinal fluid (CSF). Following affinity enrichment and trypsin digestion of CSF collected from a neurologically healthy donor, we identified several αS-derived peptides by mass spectrometry. The concentration of αS amounted to < 0.001% of the CSF proteome. We then built, validated and optimized a sandwich-type, enzyme-linked immunoadsorbent assay (ELISA) to measure total αS levels in unconcentrated CSF. In a cross-sectional study of 100 living donors, we examined cell-free CSF samples from subjects clinically diagnosed with advanced PD, dementia with Lewy bodies (DLB), Alzheimer disease (AD), and a group of non-neurodegenerative disease controls (NCO). In these four groups the CSF αS concentrations ranged from 0.8 to 16.2 pg/μl. Mean CSF αS values were lower in donors with a primary synucleinopathy (PD, DLB: n = 57) than in the other two groups (AD, NCO: n = 35; p = 0.025). By contrast, living Creutzfeldt-Jakob disease patients showed markedly elevated CSF αS levels (n = 8; mean, 300 pg/μl; p < 0.001). Our results unequivocally confirm the presence of αS in adult human CSF. In a first feasibility study employing a novel ELISA, we found relatively low CSF αS concentrations in subjects with parkinsonism linked to synucleinopathy, PD and DLB. In definite prion disease cases, we recorded a marked rise in total CSF αS resulting from rapid cell death. Our results will likely aid future biomarker explorations in neurodegenerative conditions and facilitate target validation studies.

Original languageEnglish
Pages (from-to)315-325
Number of pages11
JournalExperimental Neurology
Volume213
Issue number2
DOIs
Publication statusPublished - Oct 2008
Externally publishedYes

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Keywords

  • α-synuclein
  • Biomarker
  • Cerebrospinal fluid
  • Dementia
  • ELISA
  • Mass spectrometry
  • Parkinsonism

ASJC Scopus subject areas

  • Neurology
  • Neuroscience(all)

Cite this

Mollenhauer, B., Cullen, V., Kahn, I., Krastins, B., Outeiro, T. F., Pepivani, I., Ng, J., Schulz-Schaeffer, W., Kretzschmar, H. A., McLean, P. J., Trenkwalder, C., Sarracino, D. A., VonSattel, J. P., Locascio, J. J., Ali El-Agnaf, O., & Schlossmacher, M. G. (2008). Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration. Experimental Neurology, 213(2), 315-325. https://doi.org/10.1016/j.expneurol.2008.06.004