EAGER: Development of a Fluorescent Reporter for Protein-Membrane Interactions

The interaction of proteins with membranes in brain cells is fundamental to how the brain works. These interactions are also a contributing factor to various neurologic diseases such as Parkinson’s disease. This project involves the development of an imaging method to study protein-membrane interactions in brain cells. This research will enable studies of the underlying cause of Parkinson’s disease, which may result from the clumping of a protein called alpha-synuclein. The clumped protein then spreads from brain cell to brain cell throughout the brain. Although the alpha-synuclein clumping process is believed to play a role in Parkinson’s disease, how this clumping occurs is not fully understood. One possible mechanism involves interactions of alpha-synuclein with certain membranes in brain cells. With the development of the proposed imaging method, it will become possible to study the entire process in live brain cells and hence develop further understanding of Parkinson’s disease. The project involves two graduate students. In addition, an interactive learning module for seventh grade math students will be presented at a local school. This project is laying the groundwork for an imaging scheme using tethered fluorescence resonance energy transfer reporters to allow interrogation of membrane-induced alpha-synuclein aggregation in live, cultured neurons. The ultimate goal is to study intracellular lipid vesicles involved in endolysosomal transport, as well as synaptic vesicles, to address the key neuroscience question of which types of membrane-bound entities serve as sites of alpha-synuclein binding and aggregation in live neuron models of Parkinson’s disease. To enable this line of research, this project has two objectives: (i) realizing a method to monitor alpha-synuclein -lipid membrane association in live neurons, and (ii) developing a technique to monitor membrane-mediated alpha-synuclein aggregation in live neurons. Success in the development of this optical sensing method to monitor alpha-synuclein aggregation and prion-like spread will enable understanding of Parkinson’s disease that could be extended to animal model investigations. At a more general level, this project could have a broad impact on the understanding of a variety of human diseases, protein-membrane interactions, and the molecular underpinnings of brain function. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.