A human GBA-L444P transgene drives early and persistent dopamine neurotransmission deficits and alpha-synuclein pathology in a mouse model of early Parkinson’s disease
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Background
Heterozygous mutations in the GBA1 gene encoding the enzyme glucocerebrosidase (GCase) represent the most common genetic risk factor for developing Parkinson’s disease (PD). The underlying mechanisms by which GBA1 mutations lead to PD through both loss- and gain-of-function effects remain unclear. There is a strong rationale for the generation and characterisation of a humanised GBA1 mouse model to allow the effect of GBA1 mutations on GCase function to be studied within the context of the human protein.
Methods
We have generated novel humanised mutant GBA-L444P and wild type GBA-WT mouse models using BAC recombineering and site-specific integration allowing the incorporation of the whole GBA1 locus as a transgene, including the endogenous promoter, all exons and introns, and flanking regions. Our experimental design crossed each GBA1 transgene onto a Gba +/- background and included Gba +/- littermate controls in our cohorts, allowing us to explore both the loss- and gain-of-function of GBA1 mutations. We have carried out “deep phenotyping” to characterise these mice by biochemical, stereological and behavioural testing, and assess dopamine release and content using fast-scan cyclic voltammetry and high performance liquid chromatography.
Results
The GBA-L444P mice showed a significant reduction in GCase activity by 18 months of age and preferentially expressed a high molecular weight form of the GCase protein, likely due to retention in the ER and aberrant glycosylation. The GBA-L444P, but not Gba +/- , mice demonstrated an early and persistent reduction in dorsal striatal dopamine release in the absence of any dopaminergic cell loss or deficits in dopamine synthesis or reuptake, compared to human wild-type controls. GBA-L444P and Gba +/- mice developed an accumulation of oligomeric α-synuclein pathology, but only GBA-L444P mice demonstrated subtle but significant changes in behaviour.
Conclusions
The novel humanised GBA-L444P mouse model described here helps to resolve gain- or loss-of-function effects of GBA1 mutations seen in Parkinson’s as well as providing a novel set of models to investigate the human protein. Our work demonstrates that changes in dopamine release and behavioural deficits arise from a gain-of-function mechanism, whereas α-synuclein pathology arises from GCase loss-of-function.
