Towards an open resource for viral-vector mediated overexpression of alpha-synuclein in the Substantia Nigra: High-scale production and optimization of a rat Parkinson’s Disease model

JOAQUIN PARDO; JANITHA MUDANNAYAKE; MARTINO AVALLONE; MARCUS DAVIDSSON; TOMAS BJÖRKLUND

Glasgow

Congreso; Federation of European Neuroscience Societies 2020; 2020

The overexpression of human alpha synuclein (ha-Syn) in the Substantia Nigra (SN) of rodentsconstitutes a promising model of Parkinson’s Disease (PD). Genetic transfer by Adeno-AssociatedVirus (AAVs) is an excellent tool for this end. However, this methodology has not been easilyreproduced outside specialized labs.We set out to develop a standard, high scale procedure to obtain a reliable PD model, to beavailable in the near future as a vector resource.We first compared promoters utilized previously in PD models to novel potent alternatives (CBh,PGK, CMV or synapsin). They were inserted upstream of the ha-Syn gene and packaged into AAVsand the expression levels compared in the SN. Subsequently, AAVs of serotypes 5 and 6 wereprepared at high scale and injected at di!erent doses in the SN. Same-level overexpression ofBlue Fluorescent Protein was implemented as control. The motor phenotype outcome wasevaluated 6 weeks after injection.We found that CBH-driven overexpression of a-Syn was superior. An injection of 9xEXP(9)genome copies of AAV 5-CBH-aSyn in the rat SN prompted motor impairment at 6 weeks postinjection.Thus, a-Syn animals performed poorly in the corridor and drug-induced rotametertasks (p