Fluorescence Zinc BioSensing and Imaging Technology

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This is a competing renewal application. The overall thrust of the project is to develop novel fluorescent sensing and imaging technology for understanding the biology of zinc, with a view to developing novel and improved diagnostics and therapies for human diseases. Evidence of zinc’s role(s) in stroke, Alzheimer’s disease, epilepsy, long term memory, gastrointestinal infectious disease, and type 2 diabetes has recently emerged, but a much more detailed understanding of zinc’s involvement will be necessary in all these cases. Previously, we had developed protein-based fluorescence indicators with outstanding sensitivity, selectivity, and tunable kinetics; using this basic approach we were able for the first time to measure (in a resting cell) free zinc in the cytoplasm, and by use of an expressible indicator, in the resting mitochondrion. For the next cycle we propose to develop a palette of new ratiometric, expressible zinc indicators that will provide rapid, sensitive, and reliable zinc quantitation in many applications. We also are proposing to apply our new sensing technology to answer a question of central importance that so far has resisted other investigators: In the hippocampal formation in the brain, is synaptic zinc taken up by the post-synaptic dendritic spines? An answer to this question will help elucidate zinc’s role in learning and memory formation.