EAGER: Optical Control and Reporting of Cystosolic and Organellar pH in situ

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All forms of life rely on biochemical processes and these processes are either accelerated or inhibited according to the concentration of protons (pH) in their immediate vicinity. The pH changes that occur within nerve cells are therefore predicted to have a profound influence on the ability of nerve cells to transfer and store information. However, it is very difficult to measure or control pH in the nerve cells of live animals, and as a result the influence of pH changes on the nervous system, is difficult to test. Here, the investigators propose to use optical techniques, combined with genetically-encoded proteins, to measure and control the pH of live nerve cells in fruit flies (Drosophila melanogaster). Fluorescent proteins will be used to measure pH, and light-activated proton pumps will be used to control pH. When deployed in the same cell, these proteins will provide the means for establishing dynamic control of pH. These tools will be invaluable for investigating the extent to which the pH fluctuates in nerve cells during naturally occurring activity, and for testing the role of these pH changes in maintaining the capacity of the nerve cells to transfer and store information. Further, the development of these tools for fruit fly nerve cells will facilitate their application in other fields of the biological sciences, from mycology and plant physiology to renal physiology and oncology. Transgenic animals and plasmids generated through this project will be made available directly to the scientific research community. Information regarding the performance and application of the tools will be made available through publications in scientific journals and presentations at scientific meetings. This project will directly provide research training for an MD/PhD graduate student and training opportunities for PhD graduate students, undergraduate students, high-school students and K-12 teachers.