@article{4ca9bb684c984ba39e9d260617e194ba,
title = "Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator",
abstract = "The ability to optically image cellular transmembrane voltages at millisecond-timescale resolutions can offer unprecedented insight into the function of living brains in behaving animals. Here, we present a point mutation that increases the sensitivity of Ace2 opsin-based voltage indicators. We use the mutation to develop Voltron2, an improved chemigeneic voltage indicator that has a 65% higher sensitivity to single APs and 3-fold higher sensitivity to subthreshold potentials than Voltron. Voltron2 retained the sub-millisecond kinetics and photostability of its predecessor, although with lower baseline fluorescence. In multiple in vitro and in vivo comparisons with its predecessor across multiple species, we found Voltron2 to be more sensitive to APs and subthreshold fluctuations. Finally, we used Voltron2 to study and evaluate the possible mechanisms of interneuron synchronization in the mouse hippocampus. Overall, we have discovered a generalizable mutation that significantly increases the sensitivity of Ace2 rhodopsin-based sensors, improving their voltage reporting capability.",
keywords = "biosensors, fluorescence imaging, fluorescent proteins, genetically encoded indicators, voltage imaging",
author = "Abdelfattah, {Ahmed S.} and Jihong Zheng and Amrita Singh and Huang, {Yi Chieh} and Daniel Reep and Getahun Tsegaye and Arthur Tsang and Arthur, {Benjamin J.} and Monika Rehorova and Olson, {Carl V.L.} and Yichun Shuai and Lixia Zhang and Fu, {Tian Ming} and Milkie, {Daniel E.} and Moya, {Maria V.} and Weber, {Timothy D.} and Lemire, {Andrew L.} and Baker, {Christopher A.} and Natalie Falco and Qinsi Zheng and Grimm, {Jonathan B.} and Yip, {Mighten C.} and Deepika Walpita and Martin Chase and Luke Campagnola and Murphy, {Gabe J.} and Wong, {Allan M.} and Forest, {Craig R.} and Jerome Mertz and Economo, {Michael N.} and Turner, {Glenn C.} and Minoru Koyama and Lin, {Bei Jung} and Eric Betzig and Ondrej Novak and Lavis, {Luke D.} and Karel Svoboda and Wyatt Korff and Chen, {Tsai Wen} and Schreiter, {Eric R.} and Hasseman, {Jeremy P.} and Ilya Kolb",
note = "Funding Information: This work was funded by the Howard Hughes Medical Institute; NIH grants DP2MH129956 (A.S.A.), R01EB029171 (M.V.M, T.D.W. J.M. M.N.E.), 1F32MH129149 (M.V.M.), and 1RF1MH126882 (M.N.E.); the Searle Scholar Program (A.S.A.); the Burroughs Wellcome Fund (A.S.A.); Charles University Grant - PRIMUS/19/MED/003 and the Czech Science Foundation grant 20-25298S (O.N. M.R. C.V.L.O.); and National Science and Technology Council, Taiwan (B.-J.L. T.-W.C.). We thank the Janelia cell culture, viviarium, jET, Molecular biology, and Virus Production teams for their assistance. We thank S. Picard, K. Aswath, and S. Shrestha for help with DeepSeq. Conceptualization, A.S.A. A.M.W. T.-W.C. and E.R.S.; methodology, A.S.A. J.Z. Y.-C.H. T.D.W. A.L.L. M.C.Y. J.M. M.K. O.N. and T.-W.C.; software, B.J.A. D.E.M. A.L.L. M.C.Y. M.C. and L.C.; formal analysis, A.S.A. J.Z. A.S. Y.-C.H. B.J.A. C.V.L.O. Y.S. L.Z. T.-M.F. C.A.B. M.C. L.C. M.K. O.N. and T.-W.C.; investigation, A.S.A. J.Z. A.S. Y.-C.H. D.R. G.T. A.T. M.R. Y.S. L.Z. T.-M.F. M.V.M. T.D.W. C.A.B. M.K. O.N. J.P.H. and I.K.; resources, N.F. Q.Z. J.B.G. D.W. and C.R.F.; data curation, A.S.A. J.Z. B.J.A. C.V.L.O. Y.S. C.A.B. M.C. L.C. O.N. and I.K.; writing/visualization, A.S.A. J.Z. A.S. Y.-C.H. A.T. B.J.A. Y.S. C.A.B. M.N.E. M.K. B.-J.L. O.N. T.-W.C. E.R.S. J.P.H. and I.K.; supervision, C.R.F. G.J.M. J.M. M.N.E. G.C.T. M.K. B.-J.L. E.B. O.N. L.D.L. K.S. W.K. T.-W.C. E.R.S. J.P.H. and I.K.; project administration, A.M.W. W.K. E.R.S. J.P.H. and I.K. A.S.A. L.D.L. and E.R.S. have filed for a patent on the chemigenetic voltage indicators. I.K. and C.R.F. are co-inventors on a patent describing pipette cleaning that is licensed by Sensapex. M.C. and L.C. have performed consulting services for Sensapex. Funding Information: This work was funded by the Howard Hughes Medical Institute ; NIH grants DP2MH129956 (A.S.A.), R01EB029171 (M.V.M, T.D.W., J.M., M.N.E.), 1F32MH129149 (M.V.M.), and 1RF1MH126882 (M.N.E.); the Searle Scholar Program (A.S.A.); the Burroughs Wellcome Fund (A.S.A.); Charles University Grant - PRIMUS/19/MED/003 and the Czech Science Foundation grant 20-25298S (O.N., M.R., C.V.L.O.); and National Science and Technology Council , Taiwan (B.-J.L., T.-W.C.). We thank the Janelia cell culture, viviarium, jET, Molecular biology, and Virus Production teams for their assistance. We thank S. Picard, K. Aswath, and S. Shrestha for help with DeepSeq. Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",
year = "2023",
month = may,
day = "17",
doi = "10.1016/j.neuron.2023.03.009",
language = "English (US)",
volume = "111",
pages = "1547--1563.e9",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "10",
}