title: A Rapid, Semi-Quantitative Assay to Screen for Modulators of Alpha-Synuclein Oligomerization Ex vivo.
abstract: Alpha synuclein (αsyn) aggregates are associated with the pathogenesis of Parkinson's disease and others related disorders. Although modulation of αsyn aggregation is an attractive therapeutic target, new powerful methodologies are desperately needed to facilitate in vivo screening of novel therapeutics. Here, we describe an in vivo rodent model with the unique ability to rapidly track αsyn-αsyn interactions and thus oligomerization using a bioluminescent protein complementation strategy that monitors spatial and temporal αsyn oligomerization ex vivo. We find that αsyn forms oligomers in vivo as early as 1 week after stereotactic AAV injection into rat substantia nigra. Strikingly, although abundant αsyn expression is also detected in striatum at 1 week, no αsyn oligomers are detected at this time point. By 4 weeks, oligomerization of αsyn is detected in both striatum and substantia nigra homogenates. Moreover, in a proof-of-principle experiment, the effect of a previously described Hsp90 inhibitor known to prevent αsyn oligomer formation, demonstrates the utility of this rapid and sensitive animal model to monitor αsyn oligomerization status in the rat brain.
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Marion
authors.initials: M
authors.lastname: Delenclos
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Teodora
authors.initials: T
authors.lastname: Trendafilova
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Daryl R
authors.initials: DR
authors.lastname: Jones
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Moussaud
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Ann-Marie
authors.initials: AM
authors.lastname: Baine
authors.affiliation: Department of Neuroscience, Mayo Clinic Jacksonville, FL, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Yue
authors.affiliation: Pfizer Neuroscience Research Unit Cambridge, MA, USA.
authors.forename: Warren D
authors.initials: WD
authors.lastname: Hirst
authors.affiliation: Department of Neuroscience, Mayo ClinicJacksonville, FL, USA; Mayo Graduate School, Mayo ClinicJacksonville, FL, USA.
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: McLean
keywords: alpha-synuclein; bioluminescence; oligomers; protein-fragment complementation; viral vector model
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