MJF-14 proximity ligation assay detects early non-inclusion alpha-synuclein pathology with enhanced specificity and sensitivity
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.
Authors
- Cristine Betzer (ORCID: https://orcid.org/0000-0001-5429-3548)
- YuHong Fu (ORCID: https://orcid.org/0000-0003-4539-2039)
- Sara Elfarrash (ORCID: https://orcid.org/0000-0002-4573-3131)
- Vladimir L. Buchman (ORCID: https://orcid.org/0000-0002-7631-8352)
- Nanna Møller Jensen (ORCID: https://orcid.org/0000-0003-1177-5197)
- Mark Denham (ORCID: https://orcid.org/0000-0001-5279-5915)
- Glenda Margaret Halliday (ORCID: https://orcid.org/0000-0003-0422-8398)
- Hjalte Gram (ORCID: https://orcid.org/0000-0001-6516-5189)
- Poul Henning Jensen (ORCID: https://orcid.org/0000-0002-4439-9020)
- Silvio O. Rizzoli (ORCID: https://orcid.org/0000-0002-1667-7839)
- Lasse Reimer (ORCID: https://orcid.org/0000-0002-6996-3185)
- Zagorka Vitic (ORCID: https://orcid.org/0009-0002-5129-841X)
- Donatus Krah (ORCID: https://orcid.org/0009-0003-1151-7726)
- Ali H. Shaib (ORCID: https://orcid.org/0009-0007-2656-3820)
- Hongyun Li (ORCID: https://orcid.org/0000-0003-3332-3132)
Institutions
- The University of Sydney (AU)
- Mansoura University (EG)
- Aarhus University (DK)
- Danish Technological Institute (DK)
- Universitätsmedizin Göttingen (DE)
- University of Göttingen (DE)
- Cardiff University (GB)
- Neuroscience Research Australia (AU)
Publication Details
- Journal
- npj Parkinson s Disease
- Published
- 2024-11-29
- DOI
- https://doi.org/10.1038/s41531-024-00841-9
- Citations
- 15
- Primary Topic
- Parkinson's Disease Mechanisms and Treatments
- Type
- article
- Field-Weighted Citation Impact
- 5.54
Funders
- Michael J. Fox Foundation for Parkinson's Research
- Aarhus Universitet
- Parkinsonforeningen
- Neuroscience Research Australia
- Deutsche Forschungsgemeinschaft
- University of Sydney
- Lundbeckfonden
- H. Lundbeck A/S
- Parkinson Vereniging
- National Institutes of Health
- National Health and Medical Research Council