Objective/Rationale:
The cause of Parkinson’s disease (PD) is unknown. However, it has become clear that the abnormal aggregation of a neuronal protein called alpha-synuclein has an important role. What causes alpha-synuclein to aggregate in PD is unknown, but several environmental and genetic factors have been implicated. The purpose of this study is to develop a novel pre-clinical model in which alpha-synuclein aggregation can be directly measured and monitored in response to these putative PD-initiating factors.
Project Description:
The goal of this project is to create a novel transgenic model: the “BiSyn” (Bimolecular Fluorescence Complementation-Synuclein) model, in which alpha-synuclein aggregation can be directly visualized in intact tissues and monitored in real time in cells. We will use a novel transgenic technology called “bimolecular fluorescence complementation.” Two full-length constructs of alpha-synuclein are created, one fused to the n-terminal half of a fluorescent protein (Venus) and the other to the C-terminal half. Neither of these constructs is fluorescent when expressed separately or when co-expressed in the absence of aggregation. However, upon aggregation, proximity of the respective N- and C-terminal Venus fragments produces visible fluorescence. We have established model stem cell lines stably transfected with the BiSyn construct and we will utilize these to create the BiSyn model.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
The abnormal aggregation of alpha-synuclein is central to PD pathogenesis. Identifying the factors that initiate alpha-synuclein aggregation has profound implications for our understanding of PD etiology. Conversely, discovering agents capable of preventing or reversing aggregation could lead to effective, disease-modifying treatments. The BiSyn model will provide a valuable tool in both respects and thereby has important implications both with respect to the understanding of the pathogenesis and to the treatment of PD.
Anticipated Outcome:
The creation of the BiSyn model will provide researchers with a novel tool that can be exploited to quantitatively evaluate the efficacy of candidate PD therapeutics and validate current and future models of PD. Moreover, in light of increasing evidence for an important role for environmental agents in initiating PD pathogenesis, it will also provide an attractive platform to interrogate candidate PD-inducing pathogens and/or toxins.