Spying on Neuronal Membrane Potential with Genetically Targetable Voltage Indicators

Vincent GrenierEvan W Miller


Methods for optical measurement of voltage dynamics in living cells are attractive because they provide spatial resolution surpassing traditional electrode-based measurements and temporal resolution exceeding that of widely-used Ca2+-imaging. Chemically-synthesized voltage-sensitive dyes that use photoinduced electron transfer (PeT) as a voltage-sensing trigger offer high voltage sensitivity and fast response kinetics, but targeting chemical indicators to specific cells remains an outstanding challenge. Here, we present a new family of readily functionalizable, fluorescein-based voltage sensitive fluorescent dyes (sarcosine-VoltageFluors) that can be covalently attached to a genetically-encoded cell surface receptor to achieve voltage imaging from genetically defined neurons. We synthesized four new VoltageFluor derivatives that possess carboxylic acid functionality for simple conjugation to flexible tethers. The best of this new group of dyes was conjugated via a polyethyleneglycol (PEG) linker to a small peptide (SpyTag, 13 amino acids) that directs binding and formation of a covalent bond with its binding partner, SpyCatcher (15 kDa). The new VoltageSpy dyes effectively label cells expressing cell-surface SpyCatcher, display...Continue Reading

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