Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator.
biosensors
fluorescence imaging
fluorescent proteins
genetically encoded indicators
voltage imaging
Journal
Neuron
ISSN: 1097-4199
Titre abrégé: Neuron
Pays: United States
ID NLM: 8809320
Informations de publication
Date de publication:
17 05 2023
17 05 2023
Historique:
received:
06
12
2022
revised:
15
02
2023
accepted:
07
03
2023
pmc-release:
17
05
2024
medline:
22
5
2023
pubmed:
5
4
2023
entrez:
4
4
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 37015225
pii: S0896-6273(23)00205-2
doi: 10.1016/j.neuron.2023.03.009
pmc: PMC10280807
mid: NIHMS1883979
pii:
doi:
Substances chimiques
Rhodopsin
9009-81-8
Angiotensin-Converting Enzyme 2
EC 3.4.17.23
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1547-1563.e9Subventions
Organisme : NIMH NIH HHS
ID : DP2 MH129956
Pays : United States
Organisme : NIMH NIH HHS
ID : F32 MH129149
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS102727
Pays : United States
Organisme : NIA NIH HHS
ID : RF1 AG079269
Pays : United States
Informations de copyright
Copyright © 2023 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests 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.
Références
Cell. 2019 Dec 12;179(7):1590-1608.e23
pubmed: 31835034
J Neural Eng. 2019 Aug;16(4):046003
pubmed: 30970335
PLoS One. 2013 Oct 14;8(10):e77728
pubmed: 24155972
Nature. 1999 Nov 4;402(6757):75-9
pubmed: 10573419
Science. 2018 Apr 20;360(6386):
pubmed: 29674564
Methods Mol Biol. 2014;1183:221-42
pubmed: 25023312
PLoS One. 2013 Dec 31;8(12):e85221
pubmed: 24391999
Sci Adv. 2021 May 5;7(19):
pubmed: 33952514
Nat Rev Neurosci. 2019 Dec;20(12):719-727
pubmed: 31705060
J Neurosci. 2014 Nov 5;34(45):14984-94
pubmed: 25378164
Neuron. 2010 Feb 11;65(3):422-35
pubmed: 20159454
J Neurophysiol. 2009 Jul;102(1):636-43
pubmed: 19386760
Cell. 2020 Feb 6;180(3):521-535.e18
pubmed: 31978320
Nature. 2019 Oct;574(7778):413-417
pubmed: 31597963
Nat Methods. 2023 Jan 9;:
pubmed: 36624211
Nat Commun. 2014 Aug 13;5:4625
pubmed: 25118186
J Neurosci. 2019 Jun 19;39(25):4889-4908
pubmed: 30952812
J Mol Biol. 2004 Jan 2;335(1):297-305
pubmed: 14659758
Nat Commun. 2020 Jul 10;11(1):3444
pubmed: 32651384
J Neurophysiol. 2016 Jul 1;116(1):135-52
pubmed: 27075539
Proc Natl Acad Sci U S A. 2011 Aug 23;108(34):14133-8
pubmed: 21831834
Science. 2015 Dec 11;350(6266):1361-6
pubmed: 26586188
Elife. 2016 Mar 24;5:
pubmed: 27011354
Lab Anim (NY). 2011 May;40(5):155-60
pubmed: 21508954
Synapse. 2006 Nov;60(6):437-40
pubmed: 16881073
ACS Chem Biol. 2008 Jun 20;3(6):373-82
pubmed: 18533659
Nat Methods. 2011 Nov 27;9(1):90-5
pubmed: 22120467
Biophys J. 2013 Jan 8;104(1):51-62
pubmed: 23332058
Front Neuroinform. 2014 Jan 30;8:3
pubmed: 24523692
Biomed Eng Online. 2003 May 17;2:13
pubmed: 12801419
Nat Methods. 2019 Dec;16(12):1226-1232
pubmed: 31570887
Nat Methods. 2015 Mar;12(3):244-50, 3 p following 250
pubmed: 25599551
Sci Rep. 2016 Oct 11;6:35001
pubmed: 27725751
Nat Neurosci. 2005 Sep;8(9):1263-8
pubmed: 16116447
J Vis Exp. 2014 Sep 15;(91):51863
pubmed: 25286085
Proc Natl Acad Sci U S A. 2014 Sep 9;111(36):13034-9
pubmed: 25157169
Chem Sci. 2015 Mar;6(3):1701-1705
pubmed: 25705368
Neuron. 2016 May 18;90(4):810-23
pubmed: 27161527
Nat Methods. 2015 Feb;12(2):140-6
pubmed: 25532138
J R Soc Interface. 2013 May 15;10(84):20130197
pubmed: 23676894
Nature. 2019 May;569(7756):413-417
pubmed: 31043747
Science. 2022 Nov 04;378(6619):eabm8797
pubmed: 36378956
Nat Methods. 2017 Oct;14(10):987-994
pubmed: 28869757
Nat Methods. 2018 Dec;15(12):1108-1116
pubmed: 30420685
J Neurosci. 2011 Mar 2;31(9):3351-61
pubmed: 21368047
Cell Rep. 2019 Jan 15;26(3):802-813.e4
pubmed: 30650368
iScience. 2020 Aug 21;23(8):101363
pubmed: 32717641
Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):13222-7
pubmed: 12235359
Nat Chem Biol. 2018 Apr;14(4):352-360
pubmed: 29483642
Optica. 2019 Apr 20;6(4):389-395
pubmed: 34504902
Nat Neurosci. 2010 Nov;13(11):1433-40
pubmed: 20890294
J Neurosci. 2017 May 17;37(20):5084-5098
pubmed: 28432140
Science. 2019 Aug 16;365(6454):699-704
pubmed: 31371562
J Neurosci Methods. 2017 Nov 1;291:83-94
pubmed: 28782629
Nature. 1999 Nov 4;402(6757):72-5
pubmed: 10573418
Neuron. 2017 Oct 11;96(2):505-520.e7
pubmed: 29024669
Nat Neurosci. 2014 Jun;17(6):884-9
pubmed: 24755780
Nat Neurosci. 2006 May;9(5):608-10
pubmed: 16617340
PLoS One. 2013 Jun 19;8(6):e66959
pubmed: 23840563
J Physiol. 2017 Mar 1;595(5):1465-1477
pubmed: 27861906
PLoS Comput Biol. 2021 Apr 14;17(4):e1008806
pubmed: 33852574
Nat Methods. 2019 Jul;16(7):649-657
pubmed: 31209382
Nat Neurosci. 2016 Aug 26;19(9):1142-53
pubmed: 27571193
Nat Commun. 2014 Sep 15;5:4894
pubmed: 25222271
Biophys J. 2008 Nov 1;95(9):4384-95
pubmed: 18658225
Nat Methods. 2009 May;6(5):343-5
pubmed: 19363495
Nat Neurosci. 2014 Jun;17(6):841-50
pubmed: 24747575
Nat Methods. 2014 Aug;11(8):825-33
pubmed: 24952910
Genome Res. 2014 Dec;24(12):2033-40
pubmed: 25079858
Neuron. 2013 Oct 16;80(2):350-7
pubmed: 24139040
ACS Cent Sci. 2019 Sep 25;5(9):1602-1613
pubmed: 31572787