title: All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins.
abstract: All-optical electrophysiology-spatially resolved simultaneous optical perturbation and measurement of membrane voltage-would open new vistas in neuroscience research. We evolved two archaerhodopsin-based voltage indicators, QuasAr1 and QuasAr2, which show improved brightness and voltage sensitivity, have microsecond response times and produce no photocurrent. We engineered a channelrhodopsin actuator, CheRiff, which shows high light sensitivity and rapid kinetics and is spectrally orthogonal to the QuasArs. A coexpression vector, Optopatch, enabled cross-talk-free genetically targeted all-optical electrophysiology. In cultured rat neurons, we combined Optopatch with patterned optical excitation to probe back-propagating action potentials (APs) in dendritic spines, synaptic transmission, subcellular microsecond-timescale details of AP propagation, and simultaneous firing of many neurons in a network. Optopatch measurements revealed homeostatic tuning of intrinsic excitability in human stem cell-derived neurons. In rat brain slices, Optopatch induced and reported APs and subthreshold events with high signal-to-noise ratios. The Optopatch platform enables high-throughput, spatially resolved electrophysiology without the use of conventional electrodes.
authors: 1] Applied Physics Program, School of Engineering and Applied Science, Harvard University, Cambridge, Massachusetts, USA. [2].
authors: Daniel R
authors: DR
authors: Hochbaum
authors: 1] Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada. [2].
authors: Yongxin
authors: 0000000341885725
authors: Y
authors: Zhao
authors: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Samouil L
authors: SL
authors: Farhi
authors: 1] The MIT Media Laboratory, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA. [3] Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA. [4] McGovern Institute for Brain Research, MIT, Cambridge, Massachusetts, USA.
authors: Nathan
authors: N
authors: Klapoetke
authors: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Christopher A
authors: CA
authors: Werley
authors: Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Vikrant
authors: V
authors: Kapoor
authors: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Peng
authors: P
authors: Zou
authors: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Joel M
authors: JM
authors: Kralj
authors: Department of Physics, Harvard University, Cambridge, Massachusetts, USA.
authors: Dougal
authors: D
authors: Maclaurin
authors: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Niklas
authors: N
authors: Smedemark-Margulies
authors: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Jessica L
authors: JL
authors: Saulnier
authors: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Gabriella L
authors: GL
authors: Boulting
authors: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Christoph
authors: C
authors: Straub
authors: 1] The MIT Media Laboratory, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA. [3] Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA. [4] McGovern Institute for Brain Research, MIT, Cambridge, Massachusetts, USA.
authors: Yong Ku
authors: YK
authors: Cho
authors: Institute of Botany, Cologne Biocenter, University of Cologne, Cologne, Germany.
authors: Michael
authors: M
authors: Melkonian
authors: 1] Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada. [2] Department of Medicine, University of Alberta, Edmonton, Alberta, Canada. [3] Beijing Genomics Institute-Shenzhen, Shenzhen, China.
authors: Gane Ka-Shu
authors: GK
authors: Wong
authors: Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.
authors: D Jed
authors: DJ
authors: Harrison
authors: Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Venkatesh N
authors: VN
authors: Murthy
authors: 1] Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors: Bernardo L
authors: BL
authors: Sabatini
authors: 1] The MIT Media Laboratory, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA. [3] Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA. [4] McGovern Institute for Brain Research, MIT, Cambridge, Massachusetts, USA. [5].
authors: Edward S
authors: ES
authors: Boyden
authors: 1] Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada. [2].
authors: Robert E
authors: 000000030604092X
authors: RE
authors: Campbell
authors: 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Physics, Harvard University, Cambridge, Massachusetts, USA. [3] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors: Adam E
authors: AE
authors: Cohen
mesh_terms: D000818:Animals; D019020:Directed Molecular Evolution; D008322:Mammals / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D012243:Rhodopsin / Q000502:physiology*; D009435:Synaptic Transmission
chemical_list: D011994:Recombinant Proteins; D012243:Rhodopsin