Exploration of genetically encoded voltage indicators based on a chimeric voltage sensing domain

Deciphering how the brain generates cognitive function from patterns of electrical signals is one of the ultimate challenges in neuroscience. To this end, it would be highly desirable to monitor the activities of very large numbers of neurons while an animal engages in complex behaviors. Optical imaging of electrical activity using genetically encoded voltage indicators (GEVIs) has the potential to meet this challenge. Currently prevalent GEVIs are based on the voltage-sensitive fluorescent protein (VSFP) prototypical design or on the voltage-dependent state transitions of microbial opsins. We recently introduced a new VSFP design in which the voltage-sensing domain (VSD) is sandwiched between a fluorescence resonance energy transfer pair of fluorescent proteins (termed VSFP-Butterflies) and also demonstrated a series of chimeric VSD in which portions of the VSD of Ciona intestinalis voltage-sensitive phosphatase are substituted by homologous portions of a voltage-gated potassium channel subunit. These chimeric VSD had faster sensing kinetics than that of the native Ci-VSD. Here, we describe a new set of VSFPs that combine chimeric VSD with the Butterfly structure. We show that these chimeric VSFP-Butterflies can report membrane voltage oscillations of up to 200 Hz in cultured cells and report sensory evoked cortical population responses in living mice. This class of GEVIs may be suitable for imaging of brain rhythms in behaving mammalians.

[1]  W. N. Ross,et al.  A large change in dye absorption during the action potential. , 1974, Biophysical journal.

[2]  A. Grinvald,et al.  Simultaneous recording from several neurones in an invertebrate central nervous system , 1977, Nature.

[3]  R. D. Woods,et al.  Geophysical Characterization of Sites , 1994 .

[4]  A. Grinvald,et al.  Imaging Cortical Dynamics at High Spatial and Temporal Resolution with Novel Blue Voltage-Sensitive Dyes , 1999, Neuron.

[5]  T. Knöpfel,et al.  Design and characterization of a DNA‐encoded, voltage‐sensitive fluorescent protein , 2001, The European journal of neuroscience.

[6]  A. Grinvald,et al.  Interaction of sensory responses with spontaneous depolarization in layer 2/3 barrel cortex , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[7]  A. Grinvald,et al.  Spatiotemporal Dynamics of Sensory Responses in Layer 2/3 of Rat Barrel Cortex Measured In Vivo by Voltage-Sensitive Dye Imaging Combined with Whole-Cell Voltage Recordings and Neuron Reconstructions , 2003, The Journal of Neuroscience.

[8]  A. Grinvald,et al.  Spontaneously emerging cortical representations of visual attributes , 2003, Nature.

[9]  Amiram Grinvald,et al.  VSDI: a new era in functional imaging of cortical dynamics , 2004, Nature Reviews Neuroscience.

[10]  Walther Akemann,et al.  Engineering and Characterization of an Enhanced Fluorescent Protein Voltage Sensor , 2007, Neuroscience Research.

[11]  Thomas Knöpfel,et al.  In vivo calcium imaging from genetically specified target cells in mouse cerebellum , 2007, NeuroImage.

[12]  Yasushi Okamura,et al.  Improving membrane voltage measurements using FRET with new fluorescent proteins , 2008, Nature Methods.

[13]  Walther Akemann,et al.  Engineering of a Genetically Encodable Fluorescent Voltage Sensor Exploiting Fast Ci-VSP Voltage-Sensing Movements , 2008, PloS one.

[14]  F. Bezanilla,et al.  S4-based voltage sensors have three major conformations , 2008, Proceedings of the National Academy of Sciences.

[15]  Thomas Knöpfel,et al.  Red-shifted voltage-sensitive fluorescent proteins. , 2009, Chemistry & biology.

[16]  Walther Akemann,et al.  Spectrally-Resolved Response Properties of the Three Most Advanced FRET Based Fluorescent Protein Voltage Probes , 2009, PloS one.

[17]  Walther Akemann,et al.  Frontiers in Molecular Neuroscience Molecular Neuroscience Review Article Second and Third Generation Voltage-sensitive Fl Uorescent Proteins for Monitoring Membrane Potential , 2022 .

[18]  Walther Akemann,et al.  Imaging brain electric signals with genetically targeted voltage-sensitive fluorescent proteins , 2010, Nature Methods.

[19]  Walther Akemann,et al.  Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP , 2010, European Biophysics Journal.

[20]  Thomas Knöpfel,et al.  Genetically encoded optical indicators for the analysis of neuronal circuits , 2012, Nature Reviews Neuroscience.

[21]  K. Schulten,et al.  An emerging consensus on voltage-dependent gating from computational modeling and molecular dynamics simulations , 2012, The Journal of general physiology.

[22]  Walther Akemann,et al.  Genetically engineered fluorescent voltage reporters. , 2012, ACS chemical neuroscience.

[23]  Vincent A. Pieribone,et al.  Single Action Potentials and Subthreshold Electrical Events Imaged in Neurons with a Fluorescent Protein Voltage Probe , 2012, Neuron.

[24]  Thomas Knöpfel,et al.  Transfer of Kv3.1 voltage sensor features to the isolated Ci-VSP voltage-sensing domain. , 2012, Biophysical journal.

[25]  Walther Akemann,et al.  Imaging neural circuit dynamics with a voltage-sensitive fluorescent protein. , 2012, Journal of neurophysiology.

[26]  H. Mutoh,et al.  Probing neuronal activities with genetically encoded optical indicators: from a historical to a forward-looking perspective , 2013, Pflügers Archiv - European Journal of Physiology.

[27]  Michael Z. Lin,et al.  Improving FRET dynamic range with bright green and red fluorescent proteins , 2012, Nature Methods.

[28]  D. Maclaurin,et al.  Optical recording of action potentials in mammalian neurons using a microbial rhodopsin , 2011, Nature Methods.

[29]  Yasushi Okamura,et al.  Improved detection of electrical activity with a voltage probe based on a voltage‐sensing phosphatase , 2013, The Journal of physiology.

[30]  T. Knöpfel,et al.  Optogenetic reporters , 2013, Biology of the cell.

[31]  Dougal Maclaurin,et al.  Mechanism of voltage-sensitive fluorescence in a microbial rhodopsin , 2013, Proceedings of the National Academy of Sciences.

[32]  Bradley J. Baker,et al.  Fluorescent Protein Voltage Probes Derived from ArcLight that Respond to Membrane Voltage Changes with Fast Kinetics , 2013, PloS one.

[33]  D. J. Harrison,et al.  Bright and fast multi-colored voltage reporters via electrochromic FRET , 2014, Nature Communications.

[34]  F. Rodríguez-Ropero,et al.  Biophysical characterization. , 2014, Advances in experimental medicine and biology.

[35]  Michael Z. Lin,et al.  High-fidelity optical reporting of neuronal electrical activity with an ultrafast fluorescent voltage sensor , 2014, Nature Neuroscience.

[36]  Mark J. Schnitzer,et al.  Imaging neural spiking in brain tissue using FRET-opsin protein voltage sensors , 2014, Nature Communications.