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All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins.
Hochbaum, Daniel R; Zhao, Yongxin; Farhi, Samouil L; Klapoetke, Nathan; Werley, Christopher A; Kapoor, Vikrant; Zou, Peng; Kralj, Joel M; Maclaurin, Dougal; Smedemark-Margulies, Niklas; Saulnier, Jessica L; Boulting, Gabriella L; Straub, Christoph; Cho, Yong Ku; Melkonian, Michael; Wong, Gane Ka-Shu; Harrison, D Jed; Murthy, Venkatesh N; Sabatini, Bernardo L; Boyden, Edward S; Campbell, Robert E; Cohen, Adam E.
Afiliación
  • Hochbaum DR; 1] Applied Physics Program, School of Engineering and Applied Science, Harvard University, Cambridge, Massachusetts, USA. [2].
  • Zhao Y; 1] Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada. [2].
  • Farhi SL; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Klapoetke N; 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
  • Werley CA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Kapoor V; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Zou P; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Kralj JM; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Maclaurin D; Department of Physics, Harvard University, Cambridge, Massachusetts, USA.
  • Smedemark-Margulies N; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Saulnier JL; Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
  • Boulting GL; Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
  • Straub C; Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
  • Cho YK; 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
  • Melkonian M; Institute of Botany, Cologne Biocenter, University of Cologne, Cologne, Germany.
  • Wong GK; 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.
  • Harrison DJ; Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.
  • Murthy VN; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
  • Sabatini BL; 1] Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
  • Boyden ES; 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
  • Campbell RE; 1] Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada. [2].
  • Cohen AE; 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.
Nat Methods ; 11(8): 825-33, 2014 Aug.
Article en En | MEDLINE | ID: mdl-24952910
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.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Rodopsina / Mamíferos / Neuronas Límite: Animals Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2014 Tipo del documento: Article
Texto completo
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XML
PubMed Links
Buscar en Google

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Rodopsina / Mamíferos / Neuronas Límite: Animals Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2014 Tipo del documento: Article