Brain-derived and in vitro-seeded alpha-synuclein fibrils exhibit distinct biophysical profiles
- Nuffield Department of Clinical Neurosciences, Oxford Parkinson’s Disease Center, University of Oxford, United Kingdom
Figures
Figure 1
Distinct alpha-synuclein (αSyn) strains are associated with different neuropathological and clinical hallmarks of Parkinson’s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA).
αSyn misfolds and aggregates into fibrils with characteristic conformations. At the atomic level, PD and DLB strains share a ‘Lewy fold’ structure at the fibrillar core and comprise a single …
Figure 2 with 1 supplement
Alpha-synuclein (αSyn) seeding amplification assay (SAA) seeded with αSyn fibrils from Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) brains.
(A) SAA was performed with sarkosyl insoluble fractions of PD (n=3), PDD (n=3), DLB (n=3), MSA (n=3), and HC (n=3) brains. The curves represent an average of six replicates. Rec αSyn indicates an …
Figure 2—figure supplement 1
Raw data of the alpha-synuclein (αSyn) seeding amplification assay (SAA) reaction seeded with αSyn fibrils from Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), multiple system atrophy (MSA), and healthy control brains.
The curves represent the average of 6 replicates, and error bars indicate ± one standard deviation (SD). RFU, relative fluorescence unit.
Figure 3 with 1 supplement
Brain-derived and SAA alpha-synuclein (αSyn) fibrils exhibited distinct biochemical profiles.
Brain-derived and SAA αSyn fibrils were subjected to denaturation with increasing concentrations of GdnHCl (0–5 M) and to PK digestion (1 μg/ml). The antibody clone 42 (BD Biosciences) was used to …
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Figure 3—source data 1
Unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig3-data1-v1.zip
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Figure 3—source data 2
Labelled unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig3-data2-v1.zip
Figure 3—figure supplement 1
The full biochemical profiles of the brain-derived and SAA alpha-synuclein (αSyn) fibrils from all the cases.
The brain-derived (BD) and SAA αSyn fibrils from all three cases of (A) Parkinson’s disease (PD), (B) Parkinson’s disease with dementia (PDD), (C) dementia with Lewy bodies (DLB), and (D) multiple …
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Figure 3—figure supplement 1—source data 1
Unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig3-figsupp1-data1-v1.zip
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Figure 3—figure supplement 1—source data 2
Labelled unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig3-figsupp1-data2-v1.zip
Figure 4 with 1 supplement
Proteinase-K degradation patterns of the brain-derived alpha-synuclein (αSyn) fibrils.
The brain-derived αSyn fibrils from (A) Parkinson’s disease (PD), (B) Parkinson’s disease with dementia (PDD), (C) dementia with Lewy bodies (DLB), (D) multiple system atrophy (MSA), and (E) HC were …
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Figure 4—source data 1
Unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig4-data1-v1.zip
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Figure 4—source data 2
Labelled unedited western blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig4-data2-v1.zip
Figure 4—figure supplement 1
Immunohistochemical detection of alpha-synuclein (αSyn) in Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) brains.
The entorhinal cortex (PD, PDD, DLB) and striatum (MSA) were stained with total αSyn using antibody clone 42 (BD Biosciences). (A–C) PD cases 1–3. (D–F) PDD cases 1–3. (G–I) DLB cases 1-3. (J–L) MSA …
Figure 5 with 2 supplements
Transmission electron microscopy revealed different structures of brain-derived and seeding amplification assay (SAA) alpha-synuclein (αSyn) fibrils.
(A) Electron microscope image of negatively stained brain-derived (BD) and SAA fibrils from Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), and …
Figure 5—figure supplement 1
Transmission electron microscopy (TEM) images of the twisted brain-derived alpha-synuclein (αSyn) fibrils.
Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) brain-derived fibrils displayed intertwined fibrils with twists. …
Figure 5—figure supplement 2
Comparison of the length and width of the brain-derived and SAA alpha-synuclein (αSyn) fibrils.
Comparison of the (A) lengths and the (B) widths of the brain-derived and SAA αSyn fibrils. Thirty fibrils were counted for each case. Thus, a total of 90 data points were plotted (n=90). The black …
Figure 6 with 1 supplement
Brain-derived and seeding amplification assay (SAA) alpha-synuclein (αSyn) fibrils showed distinct phosphorylation patterns.
(A) Electron microscope image of brain-derived and SAA αSyn fibrils from Parkinson’s disease (PD), Parkinson’s disease with dementia (PDD), dementia with Lewy bodies (DLB), and multiple system …
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Figure 6—source data 1
Unedited slot blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig6-data1-v1.zip
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Figure 6—source data 2
Labelled unedited slot blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig6-data2-v1.zip
Figure 6—figure supplement 1
pS129 immunogold transmission electron microscopy (TEM) images of the brain-derived alpha-synuclein (αSyn) fibrils.
The brain-derived αSyn fibrils were labeled with anti-pαSyn (pS129) and imaged at low magnification to compare the relative quantities of the pαSyn fibrils between the diseases. All three cases were …
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Figure 6—figure supplement 1—source data 1
Unedited slot blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig6-figsupp1-data1-v1.zip
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Figure 6—figure supplement 1—source data 2
Labelled unedited slot blots.
- https://cdn.elifesciences.org/articles/92775/elife-92775-fig6-figsupp1-data2-v1.zip
Additional files
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Supplementary file 1
Clinicopathological summary of the patients included in this study.
The table summarizes the clinical and pathological reports of the patients included in this study. PD, Parkinson’s disease; PDD, Parkinson’s disease with dementia; DLB, dementia with Lewy body; MSA, multiple system atrophy; HC, healthy control; ECtx, entorhinal cortex; BG, basal ganglia; M, male; F, female; AD, Alzheimer’s disease; CVD, cardiovascular disease; and alpha-synuclein (αSyn).
- https://cdn.elifesciences.org/articles/92775/elife-92775-supp1-v1.pdf
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MDAR checklist
- https://cdn.elifesciences.org/articles/92775/elife-92775-mdarchecklist1-v1.docx
