Single Action Potentials and Subthreshold Electrical Events Imaged in Neurons with a Fluorescent Protein Voltage Probe

Overview

Journal Neuron

Publisher Cell Press

Specialty Neurology

Date 2012 Sep 11

PMID 22958819

Citations 259

Authors

Lei Jin

Zhou Han

Jelena Platisa

Julian R A Wooltorton

Lawrence B Cohen

Vincent A Pieribone

Affiliations

Soon will be listed here.

Abstract

Monitoring neuronal electrical activity using fluorescent protein-based voltage sensors has been limited by small response magnitudes and slow kinetics of existing probes. Here we report the development of a fluorescent protein voltage sensor, named ArcLight, and derivative probes that exhibit large changes in fluorescence intensity in response to voltage changes. ArcLight consists of the voltage-sensing domain of Ciona intestinalis voltage-sensitive phosphatase and super ecliptic pHluorin that carries the point mutation A227D. The fluorescence intensity of ArcLight A242 decreases by 35% in response to a 100 mV depolarization when measured in HEK293 cells, which is more than five times larger than the signals from previously reported fluorescent protein voltage sensors. We show that the combination of signal size and response speed of these new probes allows the reliable detection of single action potentials and excitatory potentials in individual neurons and dendrites.

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References

Lundby A, Mutoh H, Dimitrov D, Akemann W, Knopfel T . Engineering of a genetically encodable fluorescent voltage sensor exploiting fast Ci-VSP voltage-sensing movements. PLoS One. 2008; 3(6):e2514. PMC: 2429971. DOI: 10.1371/journal.pone.0002514. View

Miesenbock G, De Angelis D, Rothman J . Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins. Nature. 1998; 394(6689):192-5. DOI: 10.1038/28190. View

Perron A, Mutoh H, Launey T, Knopfel T . Red-shifted voltage-sensitive fluorescent proteins. Chem Biol. 2010; 16(12):1268-77. PMC: 2818747. DOI: 10.1016/j.chembiol.2009.11.014. View

Jin L, Baker B, Mealer R, Cohen L, Pieribone V, Pralle A . Random insertion of split-cans of the fluorescent protein venus into Shaker channels yields voltage sensitive probes with improved membrane localization in mammalian cells. J Neurosci Methods. 2011; 199(1):1-9. PMC: 3281265. DOI: 10.1016/j.jneumeth.2011.03.028. View

Brejc K, Sixma T, Kitts P, Kain S, Tsien R, Ormo M . Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein. Proc Natl Acad Sci U S A. 1997; 94(6):2306-11. PMC: 20083. DOI: 10.1073/pnas.94.6.2306. View

Elsliger M, Wachter R, Hanson G, Kallio K, Remington S . Structural and spectral response of green fluorescent protein variants to changes in pH. Biochemistry. 1999; 38(17):5296-301. DOI: 10.1021/bi9902182. View

Sakai R, Repunte-Canonigo V, Raj C, Knopfel T . Design and characterization of a DNA-encoded, voltage-sensitive fluorescent protein. Eur J Neurosci. 2001; 13(12):2314-8. DOI: 10.1046/j.0953-816x.2001.01617.x. View

Agmon N . Proton pathways in green fluorescence protein. Biophys J. 2005; 88(4):2452-61. PMC: 1305344. DOI: 10.1529/biophysj.104.055541. View

Sakai A, Nakayama M, Numata M, Takesawa S, Nakamoto M . Sodium sulfite and N-acetylcysteine: new additives to dialysate for inhibiting formation of glucose degradation products and advanced glycation end-products. Adv Perit Dial. 2001; 17:66-70. View

10.

Murata Y, Iwasaki H, Sasaki M, Inaba K, Okamura Y . Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. Nature. 2005; 435(7046):1239-43. DOI: 10.1038/nature03650. View

11.

Villalba-Galea C, Miceli F, Taglialatela M, Bezanilla F . Coupling between the voltage-sensing and phosphatase domains of Ci-VSP. J Gen Physiol. 2009; 134(1):5-14. PMC: 2712979. DOI: 10.1085/jgp.200910215. View

12.

Akemann W, Mutoh H, Perron A, Rossier J, Knopfel T . Imaging brain electric signals with genetically targeted voltage-sensitive fluorescent proteins. Nat Methods. 2010; 7(8):643-9. DOI: 10.1038/nmeth.1479. View

13.

Ilagan R, Rhoades E, Gruber D, Kao H, Pieribone V, Regan L . A new bright green-emitting fluorescent protein--engineered monomeric and dimeric forms. FEBS J. 2010; 277(8):1967-78. PMC: 2855763. DOI: 10.1111/j.1742-4658.2010.07618.x. View

14.

Shinobu A, Palm G, Schierbeek A, Agmon N . Visualizing proton antenna in a high-resolution green fluorescent protein structure. J Am Chem Soc. 2010; 132(32):11093-102. DOI: 10.1021/ja1010652. View

15.

Baker B, Mutoh H, Dimitrov D, Akemann W, Perron A, Iwamoto Y . Genetically encoded fluorescent sensors of membrane potential. Brain Cell Biol. 2008; 36(1-4):53-67. PMC: 2775812. DOI: 10.1007/s11068-008-9026-7. View

16.

Baker B, Lee H, Pieribone V, Cohen L, Isacoff E, Knopfel T . Three fluorescent protein voltage sensors exhibit low plasma membrane expression in mammalian cells. J Neurosci Methods. 2006; 161(1):32-8. DOI: 10.1016/j.jneumeth.2006.10.005. View

17.

Siegel M, Isacoff E . A genetically encoded optical probe of membrane voltage. Neuron. 1997; 19(4):735-41. DOI: 10.1016/s0896-6273(00)80955-1. View

18.

Kralj J, Douglass A, Hochbaum D, Maclaurin D, Cohen A . Optical recording of action potentials in mammalian neurons using a microbial rhodopsin. Nat Methods. 2011; 9(1):90-5. PMC: 3248630. DOI: 10.1038/nmeth.1782. View

19.

Ataka K, Pieribone V . A genetically targetable fluorescent probe of channel gating with rapid kinetics. Biophys J. 2001; 82(1 Pt 1):509-16. PMC: 1302490. DOI: 10.1016/S0006-3495(02)75415-5. View

20.

Kralj J, Hochbaum D, Douglass A, Cohen A . Electrical spiking in Escherichia coli probed with a fluorescent voltage-indicating protein. Science. 2011; 333(6040):345-8. DOI: 10.1126/science.1204763. View