Fluorescent reporters of membrane potential are important tools in neuroscience, offering a non-invasive, real-time optical readout of neuronal activity. Both voltage sensitive dyes (VSDs) and genetically encoded voltage indicators (GEVIs) are available. VSDs of the VoltageFluor (VF) family report neuronal activity with high sensitivity and sub-millisecond response times, but lack the cell-type specific targetability of genetically encoded tools, placing certain constraints on their applicability in complex systems. In an effort to harness the best qualities of both VSDs and GEVIs we have developed the VoltageSpy system, wherein a VF dye is tethered to a 13mer peptide (SpyTag) which reacts covalently at the surface of mammalian cells exogenously expressing the SpyCatcher protein. We report the synthesis of a novel series of VF dyes and their SpyTag conjugates, their characterization in human embryonic kidney cells as well as imaging applications in primary culture. VoltageSpy selectively labels dissociated rat hippocampal neurons without requiring a washout step, and successfully reports evoked action potentials with high fidelity.<br/>

Genetic targeting of small molecule voltage indicators using SpyTag/SpyCatcher

253rd National Meeting (2017)

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