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

Nanna Møller Jensen; Yu Hong 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:10.1038/s41531-024-00841-9

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

Nanna Møller Jensen
, Yu Hong 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

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

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.

Original language	English
Article number	227
Journal	npj Parkinson's Disease
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41531-024-00841-9
State	Published - 1 Dec 2024
Externally published	Yes

ASJC Scopus subject areas

Neurology
Clinical Neurology
Cellular and Molecular Neuroscience

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Jensen, N. M., Fu, Y. H., Betzer, C., Li, H., Elfarrash, S., Shaib, A. H., Krah, D., Vitic, Z., Reimer, L., Gram, H., Buchman, V., Denham, M., Rizzoli, S. O., Halliday, G. M., & Jensen, P. H. (2024). MJF-14 proximity ligation assay detects early non-inclusion alpha-synuclein pathology with enhanced specificity and sensitivity. npj Parkinson's Disease, 10(1), Article 227. https://doi.org/10.1038/s41531-024-00841-9

@article{1e4ac0a911ad45e58bb7639446e33777,

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

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{\textquoteright}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.",

author = "Jensen, \{Nanna M{\o}ller\} and Fu, \{Yu Hong\} and Cristine Betzer and Hongyun Li and Sara Elfarrash and Shaib, \{Ali H.\} and Donatus Krah and Zagorka Vitic and Lasse Reimer and Hjalte Gram and Vladimir Buchman and Mark Denham and Rizzoli, \{Silvio O.\} and Halliday, \{Glenda M.\} and Jensen, \{Poul Henning\}",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2024.",

year = "2024",

month = dec,

day = "1",

doi = "10.1038/s41531-024-00841-9",

language = "English",

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Jensen, NM, Fu, YH, Betzer, C, Li, H, Elfarrash, S, Shaib, AH, Krah, D, Vitic, Z, Reimer, L, Gram, H, Buchman, V, Denham, M, Rizzoli, SO, Halliday, GM & Jensen, PH 2024, 'MJF-14 proximity ligation assay detects early non-inclusion alpha-synuclein pathology with enhanced specificity and sensitivity', npj Parkinson's Disease, vol. 10, no. 1, 227. https://doi.org/10.1038/s41531-024-00841-9

TY - JOUR

T1 - MJF-14 proximity ligation assay detects early non-inclusion alpha-synuclein pathology with enhanced specificity and sensitivity

AU - Jensen, Nanna Møller

AU - Fu, Yu Hong

AU - Betzer, Cristine

AU - Li, Hongyun

AU - Elfarrash, Sara

AU - Shaib, Ali H.

AU - Krah, Donatus

AU - Vitic, Zagorka

AU - Reimer, Lasse

AU - Gram, Hjalte

AU - Buchman, Vladimir

AU - Denham, Mark

AU - Rizzoli, Silvio O.

AU - Halliday, Glenda M.

AU - Jensen, Poul Henning

PY - 2024/12/1

Y1 - 2024/12/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85211169326

U2 - 10.1038/s41531-024-00841-9

DO - 10.1038/s41531-024-00841-9

M3 - Article

AN - SCOPUS:85211169326

SN - 2373-8057

VL - 10

JO - npj Parkinson's Disease

JF - npj Parkinson's Disease

IS - 1

M1 - 227

ER -

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

Abstract

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