Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator. Abdelfattah, A. S., Zheng, J., Singh, A., Huang, Y., Reep, D., Tsegaye, G., Tsang, A., Arthur, B. J., Rehorova, M., Olson, C. V. L., Shuai, Y., Zhang, L., Fu, T., Milkie, D. E., Moya, M. V., Weber, T. D., Lemire, A. L., Baker, C. A., Falco, N., Zheng, Q., Grimm, J. B., Yip, M. C., Walpita, D., Chase, M., Campagnola, L., Murphy, G. J., Wong, A. M., Forest, C. R., Mertz, J., Economo, M. N., Turner, G. C., Koyama, M., Lin, B., Betzig, E., Novak, O., Lavis, L. D., Svoboda, K., Korff, W., Chen, T., Schreiter, E. R., Hasseman, J. P., & Kolb, I. Neuron, 111(10):1547–1563.e9, May, 2023.
doi  abstract   bibtex   
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.
@article{abdelfattah_sensitivity_2023,
	title = {Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator},
	volume = {111},
	issn = {1097-4199},
	doi = {10.1016/j.neuron.2023.03.009},
	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.},
	language = {eng},
	number = {10},
	journal = {Neuron},
	author = {Abdelfattah, Ahmed S. and Zheng, Jihong and Singh, Amrita and Huang, Yi-Chieh and Reep, Daniel and Tsegaye, Getahun and Tsang, Arthur and Arthur, Benjamin J. and Rehorova, Monika and Olson, Carl V. L. and Shuai, Yichun and Zhang, Lixia 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 Falco, Natalie and Zheng, Qinsi and Grimm, Jonathan B. and Yip, Mighten C. and Walpita, Deepika and Chase, Martin and Campagnola, Luke and Murphy, Gabe J. and Wong, Allan M. and Forest, Craig R. and Mertz, Jerome and Economo, Michael N. and Turner, Glenn C. and Koyama, Minoru and Lin, Bei-Jung and Betzig, Eric and Novak, Ondrej and Lavis, Luke D. and Svoboda, Karel and Korff, Wyatt and Chen, Tsai-Wen and Schreiter, Eric R. and Hasseman, Jeremy P. and Kolb, Ilya},
	month = may,
	year = {2023},
	pmid = {37015225},
	pmcid = {PMC10280807},
	keywords = {Action Potentials, Angiotensin-Converting Enzyme 2, Animals, Mice, Mutation, Neurons, Rhodopsin, biosensors, fluorescence imaging, fluorescent proteins, genetically encoded indicators, gevi, voltage imaging},
	pages = {1547--1563.e9},
}
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