Characterization of rAAV2/8 viral vector-based expression of ɑ-synuclein in oligodendrocytes in vivo
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1
KU Leuven, Department of Neurosciences, Belgium
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2
KU Leuven, LVVC - Leuven Viral Vector Core, Belgium
Various age-related neurodegenerative diseases are characterized by aberrant protein folding in neurons and oligodendrocytes. Multiple System atrophy (MSA) belongs to the group of synucleinopathies and is associated with autonomic dysfunction, parkinsonism, cerebellar ataxia, pyramidal symptoms and deposition of misfolded ɑ-synuclein protein (ɑSyn) in oligodendrocytes. In MSA, ɑSyn pathology is apparent in striatum and cerebellum, which has led to a subdivision into a parkinsonian (MSA-P) and cerebellar phenotype (MSA-C), respectively. Selective expression of ɑSyn in transgenic rodent models has recapitulated many of the features observed in MSA patients such as glial cytoplasmic inclusions (GCIs), neurotoxicity and behavioral deficits albeit at very old age (1). All these observations are pointing towards a central role of ɑSyn-induced oligodendropathy in MSA. However, transgenic animal models are slow with a spatiotemporal restriction of ɑSyn expression and pathology. Generating an acute viral vector-based model of MSA via expression of ɑSyn in oligodendroglial cells could therefore have several new advantages, such as modulating expression levels and its expression site. In this work the general aim is therefore to generate an MSA viral vector model based on ɑSyn expression in striatal oligodendrocytes.
By using viral vector technology we first assessed GFP expression in oligodendroglial cells. We injected recombinant adeno-associated viral vector serotype 2/8 (rAAV2/8) in rat striatum and compared the expression specificity, efficiency and expression pattern of different oligodendroglial promoters, including MAG, MBP and CMVieMBP (2,3). Using equal viral vectors titers we assessed the transduced area and GFP expression pattern in oligodendrocytes, neurons and astrocytes after 2 weeks of injection. We found that different promoters had different expression specificities. Using the MBP promoter results in more than 90% expression specificity in striatal oligodendrocytes (3). We therefore subsequently selected rAAV2/8-MBP to overexpress ɑSyn in rat striatum. We assessed its capacities to induce oligodendroglial or neuronal alterations by performing behavioral and immunohistological analysis as readout for striatonigral vulnerability 1 and 2 months after injection. In addition, we performed biochemical analysis of oligodendroglial, neuronal and synucleinopathy markers. Within this relatively short timeframe, our results indicate progressive accumulation of pathological ɑSyn at different time points but in the absence of clear oligo- or neuropathological alterations.
Acknowledgements
F.B. is a Strategic Basic Research (SB) doctoral fellow from Flemish Fund for Scientific Research (FWO Vlaanderen).
References
(1) Fellner, L., Wenning, G.K. & Stefanova, N., Curr Top Behav Neurosci 22, 369-93 (2015)
(2) Jonquieres, G. et al, Frontiers in Molecular Neuroscience 9:13 (2016)
(3) Lawlor, P.A., Bland, R.J., Mouravlev, A., Young, D. & During, M.J., Mol Ther 17, 1692-702 (2009)
Keywords:
Multiple System Atrophy,
synucleinopathies,
rAAV,
viral vectors,
oligodendrocytes
Conference:
12th National Congress of the Belgian Society for Neuroscience, Gent, Belgium, 22 May - 22 May, 2017.
Presentation Type:
Poster Presentation
Topic:
Disorders of the Nervous System
Citation:
De Brito
F,
Peelaerts
W,
Van Den Haute
C and
Baekelandt
V
(2019). Characterization of rAAV2/8 viral vector-based expression of ɑ-synuclein in oligodendrocytes in vivo.
Front. Neurosci.
Conference Abstract:
12th National Congress of the Belgian Society for Neuroscience.
doi: 10.3389/conf.fnins.2017.94.00068
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Received:
24 Apr 2017;
Published Online:
25 Jan 2019.
*
Correspondence:
Miss. Filipa De Brito, KU Leuven, Department of Neurosciences, Leuven, 3000, Belgium, filipa.brito@kuleuven.be
Prof. Veerle Baekelandt, KU Leuven, LVVC - Leuven Viral Vector Core, Leuven, Flanders, 3000, Belgium, veerle.baekelandt@kuleuven.be