npj Parkinson's Disease (Nov 2024)

MJF-14 proximity ligation assay detects early non-inclusion alpha-synuclein pathology with enhanced specificity and sensitivity

  • Nanna Møller Jensen,
  • YuHong Fu,
  • Cristine Betzer,
  • Hongyun Li,
  • Sara Elfarrash,
  • Ali H. Shaib,
  • Donatus Krah,
  • Zagorka Vitic,
  • Lasse Reimer,
  • Hjalte Gram,
  • Vladimir Buchman,
  • Mark Denham,
  • Silvio O. Rizzoli,
  • Glenda M. Halliday,
  • Poul Henning Jensen

DOI
https://doi.org/10.1038/s41531-024-00841-9
Journal volume & issue
Vol. 10, no. 1
pp. 1 – 21

Abstract

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Abstract α-Synuclein proximity ligation assay (PLA) has proved a sensitive technique for detection of non-Lewy body α-synuclein aggregate pathology. Here, we describe the MJF-14 PLA, a new PLA towards aggregated α-synuclein with unprecedented specificity, using the aggregate-selective α-synuclein antibody MJFR-14-6-4-2 (hereafter MJF-14). Signal in the assay correlates with α-synuclein aggregation in cell culture and human neurons, induced by α-synuclein overexpression or pre-formed fibrils. Co-labelling of MJF-14 PLA and pS129-α-synuclein immunofluorescence in post-mortem cases of dementia with Lewy bodies shows that while the MJF-14 PLA reveals extensive non-inclusion pathology, it is not sensitive towards pS129-α-synuclein-positive Lewy bodies. In Parkinson’s disease brain, direct comparison of PLA and immunohistochemistry with the MJF-14 antibody shows widespread α-synuclein pathology preceding the formation of conventional Lewy pathology. In conclusion, we introduce an improved α-synuclein aggregate PLA to uncover abundant non-inclusion pathology, which deserves future validation with brain bank resources and in different synucleinopathies.