title: Genetically Targeted All-Optical Electrophysiology with a Transgenic Cre-Dependent Optopatch Mouse.
abstract: Recent advances in optogenetics have enabled simultaneous optical perturbation and optical readout of membrane potential in diverse cell types. Here, we develop and characterize a Cre-dependent transgenic Optopatch2 mouse line that we call Floxopatch. The animals expressed a blue-shifted channelrhodopsin, CheRiff, and a near infrared Archaerhodopsin-derived voltage indicator, QuasAr2, via targeted knock-in at the rosa26 locus. In Optopatch-expressing animals, we tested for overall health, genetically targeted expression, and function of the optogenetic components. In offspring of Floxopatch mice crossed with a variety of Cre driver lines, we observed spontaneous and optically evoked activity in vitro in acute brain slices and in vivo in somatosensory ganglia. Cell-type-specific expression allowed classification and characterization of neuronal subtypes based on their firing patterns. The Floxopatch mouse line is a useful tool for fast and sensitive characterization of neural activity in genetically specified cell types in intact tissue.
  
  
  
  Optical recordings of neural activity offer the promise of rapid and spatially resolved mapping of neural function. Calcium imaging has been widely applied in this mode, but is insensitive to the details of action potential waveforms and subthreshold events. Simultaneous optical perturbation and optical readout of single-cell electrical activity ("Optopatch") has been demonstrated in cultured neurons and in organotypic brain slices, but not in acute brain slices or in vivo Here, we describe a transgenic mouse in which expression of Optopatch constructs is controlled by the Cre-recombinase enzyme. This animal enables fast and robust optical measurements of single-cell electrical excitability in acute brain slices and in somatosensory ganglia in vivo, opening the door to rapid optical mapping of neuronal excitability.
authors: Department of Chemistry and Chemical Biology and.
authors: Shan
authors: S
authors: Lou
authors: Department of Chemistry and Chemical Biology and.
authors: Yoav
authors: Y
authors: Adam
authors: Department of Chemistry and Chemical Biology and.|Department of Physics, Harvard University, Cambridge, Massachusetts 02138.
authors: Eli N
authors: 0000-0002-8896-2911
authors: EN
authors: Weinstein
authors: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115.
authors: Erika
authors: E
authors: Williams
authors: Department of Chemistry and Chemical Biology and.
authors: Katherine
authors: K
authors: Williams
authors: Department of Chemistry and Chemical Biology and.
authors: Vicente
authors: V
authors: Parot
authors: Department of Chemistry and Chemical Biology and.
authors: Nikita
authors: 0000-0002-8037-7996
authors: N
authors: Kavokine
authors: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115.
authors: Stephen
authors: S
authors: Liberles
authors: Allen Institute for Brain Science, Seattle, Washington 98107, and.
authors: Linda
authors: L
authors: Madisen
authors: Allen Institute for Brain Science, Seattle, Washington 98107, and.
authors: Hongkui
authors: H
authors: Zeng
authors: Department of Chemistry and Chemical Biology and cohen@chemistry.harvard.edu.|Department of Physics, Harvard University, Cambridge, Massachusetts 02138.|Howard Hughes Medical Institute, Chevy Chase, Maryland 20815.
authors: Adam E
authors: AE
authors: Cohen
mesh_terms: D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D018390:Gene Targeting; D019426:Integrases / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D062308:Optogenetics / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics; D056969:Voltage-Sensitive Dye Imaging / Q000379:methods*
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; C045073:Cre recombinase; D019426:Integrases
keywords: optogenetics; optopatch; transgenic mice; voltage imaging