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Verchere, J.

Publications and source records attributed to Verchere, J..

3 recordsLinked to original sources

1.94 Angstrom structure of synthetic alpha-synuclein fibrils seeding MSA neuropathology

Summary ParagraphMultiple system atrophy (MSA) is a rapidly progressive neurodegenerative disease of unknown cause, typically affecting individuals aged 50-60 and leading to death within a decade1-3. It is characterized by glial cytoplasmic inclusions (GCIs) composed of fibrillar alpha-synuclein (aSyn)4-8, whose formation shows parallels with prion propagation9,10. While fibrils extracted from MSA brains have been structurally characterized11, their ability to replicate in a "protein-only" manner has been questioned12, and their capacity to induce GCIs in vivo remains unexplored. By contrast, the synthetic fibril strain 1B13,14, assembled from recombinant human aSyn, self-replicates in vitro and induces GCIs in mice15 - suggesting direct relevance to MSA - but awaited scrutiny at an atomic scale. Here, we report high-resolution structural analyses of 1B fibrils and of fibrils extracted from diseased mice injected with 1B that developed GCIs (1BP). We show in vivo that conformational templating enables fibril strain replication, resulting in MSA-like inclusion pathology. Remarkably, the structures of 1B and 1BP are highly similar and mimic the fold of aSyn observed in one protofilament of fibrils isolated from MSA patients11. Moreover, reinjection of crude mouse brain homogenates containing 1BP into new mice reproduces the same MSA-like pathology induced by the parent synthetic seed 1B. Our findings identify 1B as a synthetic pathogen capable of self-replication in vivo and reveal structural features of 1B/1BP that may underlie MSA pathology, offering insights for therapeutic strategies.

neuroscience↗

Seeded propagation of alpha-synuclein aggregation in mouse brain using protein misfolding cyclic amplification

Alpha-synuclein (-syn) protein aggregation is associated with several neurodegenerative disorders collectively referred to as synucleinopathies, including Parkinson disease. We used protein misfolding cyclic amplification (PMCA) to study -syn aggregation in brain homogenates of wild-type or transgenic mice expressing normal (D line) or A53T mutant (M83 line) human -syn. We found that sonication-incubation cycles of M83 mouse brain gradually produce large quantities of SDS-resistant -syn aggregates, involving both human and mouse proteins. These PMCA products, containing partially proteinase K resistant -syn species, are competent to accelerate the onset of neurological symptoms after intracerebral inoculation to young M83 mice and to seed aggregate formation of -syn following PMCA, including in D and wild-type mouse brain substrates. Our data indicate that similar to prions, PMCA can reproduce some characteristics of -syn aggregation and seeded-propagation in vitro in a complex milieu. This opens new opportunities for the molecular study of synucleinopathies.

neuroscience↗

Investigating the neuroprotective effect of AAV-mediated β-synuclein overexpression in a transgenic model of synucleinopathy

Parkinsons disease (PD) and multiple system atrophy (MSA) are neurodegenerative diseases characterized by inclusions mainly composed of -synuclein (-syn) aggregates. The objective of this study was to investigate if {beta}-synuclein ({beta}-syn) overexpression could have beneficial effects by inhibiting the aggregation of -syn. The M83 transgenic mouse is a model of synucleinopathy, which develops severe motor symptoms associated with aggregation of -syn. M83 neonate or adult mice were injected with adeno-associated virus vectors carrying the human {beta}-syn gene (AAV{beta}-syn) or green fluorescent protein gene (AAVGFP) using different injection sites. One or two months later, M83 disease was accelerated or not using brain M83 extracts from mouse (M83) or human (MSA) origins. AAV mediated {beta}-syn overexpression detected by ELISA did not delay the disease onset, regardless of the AAV injection route and of the inoculation of brain extracts. Accordingly, phosphorylated -syn levels detected by ELISA in sick mice were similar after injecting AAV{beta}-syn or AAVGFP. Instead, immunohistochemistry analysis of {beta}-syn indicated the presence of proteinase-K resistant {beta}-syn staining specifically in sick M83 mice inoculated with AAV{beta}-syn. This study indicated for the first time that {beta}-syn could form aggregates in a model of synucleinopathy when it is expressed by a viral vector.

neuroscience↗