Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator

Ahmed S. Abdelfattah; Jihong Zheng; Amrita Singh; Yi Chieh Huang; Daniel Reep; Getahun Tsegaye; Arthur Tsang; Benjamin J. Arthur; Monika Rehorova; Carl V.L. Olson; Yichun Shuai; Lixia Zhang; Tian Ming Fu; Daniel E. Milkie; Maria V. Moya; Timothy D. Weber; Andrew L. Lemire; Christopher A. Baker; Natalie Falco; Qinsi Zheng; Jonathan B. Grimm; Mighten C. Yip; Deepika Walpita; Martin Chase; Luke Campagnola; Gabe J. Murphy; Allan M. Wong; Craig R. Forest; Jerome Mertz; Michael N. Economo; Glenn C. Turner; Minoru Koyama; Bei Jung Lin; Eric Betzig; Ondrej Novak; Luke D. Lavis; Karel Svoboda; Wyatt Korff; Tsai Wen Chen; Eric R. Schreiter; Jeremy P. Hasseman; Ilya Kolb

doi:10.1016/j.neuron.2023.03.009

Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator

Ahmed S. Abdelfattah^*, Jihong Zheng, Amrita Singh, Yi Chieh Huang, Daniel Reep, Getahun Tsegaye, Arthur Tsang, Benjamin J. Arthur, Monika Rehorova, Carl V.L. Olson, Yichun Shuai, Lixia Zhang, Tian Ming Fu, Daniel E. Milkie, Maria V. Moya, Timothy D. Weber, Andrew L. Lemire, Christopher A. Baker, Natalie Falco, Qinsi ZhengJonathan B. Grimm, Mighten C. Yip, Deepika Walpita, Martin Chase, Luke Campagnola, Gabe J. Murphy, Allan M. Wong, Craig R. Forest, Jerome Mertz, Michael N. Economo, Glenn C. Turner, Minoru Koyama, Bei Jung Lin, Eric Betzig, Ondrej Novak, Luke D. Lavis, Karel Svoboda, Wyatt Korff, Tsai Wen Chen^*, Eric R. Schreiter^*, Jeremy P. Hasseman^*, Ilya Kolb^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

30 Scopus citations

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.

Original language	English
Pages (from-to)	1547-1563.e9
Journal	Neuron
Volume	111
Issue number	10
DOIs	https://doi.org/10.1016/j.neuron.2023.03.009
State	Published - 17 May 2023

Keywords

biosensors
fluorescence imaging
fluorescent proteins
genetically encoded indicators
voltage imaging

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10.1016/j.neuron.2023.03.009

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Abdelfattah, A. S., Zheng, J., Singh, A., Huang, Y. C., Reep, D., Tsegaye, G., Tsang, A., Arthur, B. J., Rehorova, M., Olson, C. V. L., Shuai, Y., Zhang, L., Fu, T. M., Milkie, D. E., Moya, M. V., Weber, T. D., Lemire, A. L., Baker, C. A., Falco, N., ... Kolb, I. (2023). Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator. Neuron, 111(10), 1547-1563.e9. https://doi.org/10.1016/j.neuron.2023.03.009

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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 = "Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",

year = "2023",

month = may,

day = "17",

doi = "10.1016/j.neuron.2023.03.009",

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Abdelfattah, AS, Zheng, J, Singh, A, Huang, YC, Reep, D, Tsegaye, G, Tsang, A, Arthur, BJ, Rehorova, M, Olson, CVL, Shuai, Y, Zhang, L, Fu, TM, Milkie, DE, Moya, MV, Weber, TD, Lemire, AL, Baker, CA, Falco, N, Zheng, Q, Grimm, JB, Yip, MC, Walpita, D, Chase, M, Campagnola, L, Murphy, GJ, Wong, AM, Forest, CR, Mertz, J, Economo, MN, Turner, GC, Koyama, M, Lin, BJ, Betzig, E, Novak, O, Lavis, LD, Svoboda, K, Korff, W, Chen, TW, Schreiter, ER, Hasseman, JP & Kolb, I 2023, 'Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator', Neuron, vol. 111, no. 10, pp. 1547-1563.e9. https://doi.org/10.1016/j.neuron.2023.03.009

TY - JOUR

T1 - Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator

AU - Abdelfattah, Ahmed S.

AU - Zheng, Jihong

AU - Singh, Amrita

AU - Huang, Yi Chieh

AU - Reep, Daniel

AU - Tsegaye, Getahun

AU - Tsang, Arthur

AU - Arthur, Benjamin J.

AU - Rehorova, Monika

AU - Olson, Carl V.L.

AU - Shuai, Yichun

AU - Zhang, Lixia

AU - Fu, Tian Ming

AU - Milkie, Daniel E.

AU - Moya, Maria V.

AU - Weber, Timothy D.

AU - Lemire, Andrew L.

AU - Baker, Christopher A.

AU - Falco, Natalie

AU - Zheng, Qinsi

AU - Grimm, Jonathan B.

AU - Yip, Mighten C.

AU - Walpita, Deepika

AU - Chase, Martin

AU - Campagnola, Luke

AU - Murphy, Gabe J.

AU - Wong, Allan M.

AU - Forest, Craig R.

AU - Mertz, Jerome

AU - Economo, Michael N.

AU - Turner, Glenn C.

AU - Koyama, Minoru

AU - Lin, Bei Jung

AU - Betzig, Eric

AU - Novak, Ondrej

AU - Lavis, Luke D.

AU - Svoboda, Karel

AU - Korff, Wyatt

AU - Chen, Tsai Wen

AU - Schreiter, Eric R.

AU - Hasseman, Jeremy P.

AU - Kolb, Ilya

PY - 2023/5/17

Y1 - 2023/5/17

N2 - 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.

AB - 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.

KW - biosensors

KW - fluorescence imaging

KW - fluorescent proteins

KW - genetically encoded indicators

KW - voltage imaging

UR - http://www.scopus.com/inward/record.url?scp=85153101791&partnerID=8YFLogxK

U2 - 10.1016/j.neuron.2023.03.009

DO - 10.1016/j.neuron.2023.03.009

M3 - Article

C2 - 37015225

AN - SCOPUS:85153101791

SN - 0896-6273

VL - 111

SP - 1547-1563.e9

JO - Neuron

JF - Neuron

IS - 10

ER -

Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator

Abstract

Keywords

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