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Bright and fast multicoloured voltage reporters via electrochromic FRET

2014; Nature Portfolio; Volume: 5; Issue: 1 Linguagem: Inglês

10.1038/ncomms5625

2041-1723

Peng Zou, Yongxin Zhao, Adam D. Douglass, Daniel R. Hochbaum, Daan Brinks, Christopher A. Werley, D. Jed Harrison, Robert E. Campbell, Adam E. Cohen,

Neuroscience and Neural Engineering

Genetically encoded fluorescent reporters of membrane potential promise to reveal aspects of neural function not detectable by other means. We present a palette of multicoloured brightly fluorescent genetically encoded voltage indicators with sensitivities from 8–13% ΔF/F per 100 mV, and half-maximal response times from 4–7 ms. A fluorescent protein is fused to an archaerhodopsin-derived voltage sensor. Voltage-induced shifts in the absorption spectrum of the rhodopsin lead to voltage-dependent nonradiative quenching of the appended fluorescent protein. Through a library screen, we identify linkers and fluorescent protein combinations that report neuronal action potentials in cultured rat hippocampal neurons with a single-trial signal-to-noise ratio from 7 to 9 in a 1 kHz imaging bandwidth at modest illumination intensity. The freedom to choose a voltage indicator from an array of colours facilitates multicolour voltage imaging, as well as combination with other optical reporters and optogenetic actuators. Genetically encoded voltage sensors are useful tools for the analysis of membrane potential and its influence on cell function. Here, the authors present a range of these sensors with varying colours for rapid and sensitive neuronal voltage imaging.