Assessment of α-Synuclein Pathology: A Study of the BrainNet Europe Consortium
Abstract
To determine the reliability of assessment of alpha-synuclein-immunoreactive (alphaS-IR) structures by neuropathologists, 28 evaluators from 17 centers of BrainNet Europe examined current methods and reproducibility of alphaS-IR evaluation using a tissue microarray (TMA) technique. Tissue microarray blocks were constructed of samples from the participating centers that contained alphaS-IR structures. Slides from these blocks were stained in each center and assessed for neuronal perikaryal inclusions, neurites, and glial cytoplasmic inclusions. The study was performed in 2 phases. First, the TMA slides were stained with the antibody of the center's choice. In this phase, 59% of the sections were of good or acceptable quality, and 4 of 9 antibodies used performed consistently. Differences in interpretation and categorization of alphaS-IR structures, however, led to differing results between the laboratories. Prior to the second phase, the neuropathologists participated in a training session on the evaluation of alphaS-IR structures. Based on the results of the first phase, selected antibodies using designated antigen retrieval methods were then applied to TMA slides in the second phase. When the designated methods of both staining and evaluation were applied, all 26 subsequently stained TMA sections evaluated were of good/acceptable quality, and a high level of concordance in the assessment of the presence or absence of specific alphaS-IR structures was achieved. A semiquantitative assessment of alphaS-IR neuronal perikaryal inclusions yielded agreements ranging from 49% to 82%, with best concordance in cortical core samples. These results suggest that rigorous methodology and dichotomized assessment (i.e. determining the presence or absence of alphaS-IR) should be applied, and that semiquantitative assessment can be recommended only for the cortical samples. Moreover, the study demonstrates that there are limitations in the scoring of alphaS-IR structures.
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Greater sensitivity for detecting Lewy bodies can be achieved with immunohistochemistry for α-synuclein compared to H&E.
“LBs may be detected in neurons of medulla, pons and midbrain with hematoxylin and eosin (H&E)-stained sections; however, greater sensitivity can be achieved with immunohistochemistry for α-synuclein and this approach is strongly preferred [3,4,13].”
From National Institute on Aging–Alzheimer’s Association guidelines for the neuropathologic assessment of Alzheimer’s disease: a practical approach (2012) · cites this paper as [3]
The Introduction (p. 125) says consensus guidelines recommend α-synuclein immunohistochemistry for synucleinopathies, supporting the preference for that approach. The paper’s staining results and Discussion assess variation among immunohistochemical methods, but do not establish that immunohistochemistry detects Lewy bodies more sensitively than H&E; searches for H&E and hematoxylin–eosin found no such comparison. (LLM, conf=0.84)
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- Authors
- Alafuzoff, I., Parkkinen, L., Al-Sarraj, S., Arzberger, T., Bell, J., Bodi, I., Bogdanovic, N., Budka, H., Ferrer, I., Gelpi, E., Gentleman, S., Giaccone, G., Kamphorst, W., King, A., Korkolopoulou, P., Kovács, G. G., Larionov, S., Meyronet, D., Monoranu, C., Morris, J., Parchi, P., Patsouris, E., Roggendorf, W., Seilhean, D., Streichenberger, N., Thal, D. R., & Kretzschmar, H.
- Affiliations
- University of Bonn (DE)
- Netherlands Brain Bank (NL)
- Corresponding author
- Alafuzoff, Irina
- ORCID
- Venue
- Journal of Neuropathology & Experimental Neurology
- Publisher
- Oxford University Press (OUP)
- Published
- 2008-02
- Volume
- 67
- Issue
- 2
- Pages
- 125-143
- ISSN
- 0022-3069 (ISSN-L)
- 1554-6578
- OpenAlex
- W2094904417
- Cited by
- 90
- References
- 2
- Citations by year
- 2026: 6 · 2025: 3 · 2024: 3 · 2023: 2 · 2021: 2 · 2019: 4 · 2018: 6 · 2017: 5 · 2016: 4 · 2015: 4 · 2014: 3 · 2013: 7 · 2012: 7
- FWCI
- 7.27
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Neuropathology & Experimental Neurology (publishedVersion)
- PubMed (publishedVersion)
- KNAW Research Portal (The Royal Netherlands Academy of Arts and Sciences) (publishedVersion)
- ResearchOnline at James Cook University (James Cook University) (submittedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Archivio istituzionale della ricerca (Alma Mater Studiorum Università di Bologna) (submittedVersion)
- KNAW research portal (Royal Academy of Art and Sciences (KNAW)) (submittedVersion)
- Topics
- Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
- Health Sciences › Medicine › Neurology › Neurological disorders and treatments
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Keywords
- alpha-synuclein, interlaboratory reproducibility, tissue microarray, glial cytoplasmic inclusions, neurites, antibody evaluation, antigen retrieval, semi-quantitative scoring, Parkinson's disease, dementia with Lewy bodies
- MeSH terms
- Adult, Aged, Aged, 80 and over, Brain, Brain Diseases, Database Management Systems, Europe, Female, Humans, Immunohistochemistry, Male, Middle Aged, Neuroglia, Neurons, Statistics, Nonparametric, Microarray Analysis, alpha-Synuclein
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