Transient changes to intracellular pH (pHi) have been linked to a variety of cell behaviors such as progression through the cell cycle and cytoskeleton remodeling. These pH-dependent cell behaviors have been characterized at the population level using non-specific tools to manipulate pHi. Therefore, very little is known about how pHi dynamics regulate single-cell behaviors. Current techniques to manipulate pHi within single cells include micropipette buffer delivery, ionophores, and proton uncagers. However, these techniques are non-specific, technically challenging, irreversible, and can be damaging to the cell. In this work, we present the characterization of an optogenetic tool to spatiotemporally manipulate pHi within single cells. We show that Archaerhodopsin (ArchT), a light-driven outward proton pump, can be used to locally and reversibly raise pHi within single cells over several minutes. Cells respond to this spatiotemporal increase in pHi by inducing local cytoskeleton remodeling and membrane ruffling. Future work will involve adapting the technique to enable pHi manipulation over the span of hours. This technique uniquely combines reversible spatiotemporal pHi control with a non-invasive and genetically encodable optogenetic tool. This technique enables the investigation of previously intractable questions in the field including how single-cell pHi dynamics drive cell polarization, actin remodeling, and migration.

Optogenetic tool to spatiotemporally manipulate intracellular pH within single cells

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.