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Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator.
Abdelfattah, Ahmed S; Zheng, Jihong; Singh, Amrita; Huang, Yi-Chieh; Reep, Daniel; Tsegaye, Getahun; Tsang, Arthur; Arthur, Benjamin J; Rehorova, Monika; Olson, Carl V L; Shuai, Yichun; Zhang, Lixia; Fu, Tian-Ming; Milkie, Daniel E; Moya, Maria V; Weber, Timothy D; Lemire, Andrew L; Baker, Christopher A; Falco, Natalie; Zheng, Qinsi; Grimm, Jonathan B; Yip, Mighten C; Walpita, Deepika; Chase, Martin; Campagnola, Luke; Murphy, Gabe J; Wong, Allan M; Forest, Craig R; Mertz, Jerome; Economo, Michael N; Turner, Glenn C; Koyama, Minoru; Lin, Bei-Jung; Betzig, Eric; Novak, Ondrej; Lavis, Luke D; Svoboda, Karel; Korff, Wyatt; Chen, Tsai-Wen; Schreiter, Eric R; Hasseman, Jeremy P; Kolb, Ilya.
Affiliation
  • Abdelfattah AS; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA. Electronic address: ahmed_abdelfattah@brown.edu.
  • Zheng J; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Singh A; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Huang YC; Institute of Neuroscience, National Yang Ming Chiao Tung University, Taipei, Taiwan.
  • Reep D; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Tsegaye G; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Tsang A; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Arthur BJ; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Rehorova M; Department of Physiology, Second Faculty of Medicine, Charles University, Prague, Czech Republic.
  • Olson CVL; Department of Physiology, Second Faculty of Medicine, Charles University, Prague, Czech Republic.
  • Shuai Y; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Zhang L; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Fu TM; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Milkie DE; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Moya MV; Department of Biomedical Engineering, Boston University, Boston, MA, USA.
  • Weber TD; Department of Biomedical Engineering, Boston University, Boston, MA, USA.
  • Lemire AL; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Baker CA; Allen Institute for Brain Science, Seattle, WA, USA.
  • Falco N; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Zheng Q; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Grimm JB; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Yip MC; George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
  • Walpita D; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Chase M; Portland, OR, USA.
  • Campagnola L; Allen Institute for Brain Science, Seattle, WA, USA.
  • Murphy GJ; Allen Institute for Brain Science, Seattle, WA, USA.
  • Wong AM; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Forest CR; George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
  • Mertz J; Department of Biomedical Engineering, Boston University, Boston, MA, USA.
  • Economo MN; Department of Biomedical Engineering, Boston University, Boston, MA, USA.
  • Turner GC; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Koyama M; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Lin BJ; Institute of Neuroscience, National Yang Ming Chiao Tung University, Taipei, Taiwan.
  • Betzig E; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; Departments of Molecular and Cell Biology and Physics, Howard Hughes Medical Institute, Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA, USA; Molecular Biophysics and Integrated Bioimagin
  • Novak O; Department of Physiology, Second Faculty of Medicine, Charles University, Prague, Czech Republic.
  • Lavis LD; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Svoboda K; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Korff W; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Chen TW; Institute of Neuroscience, National Yang Ming Chiao Tung University, Taipei, Taiwan. Electronic address: chentw@nycu.edu.tw.
  • Schreiter ER; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA. Electronic address: schreitere@janelia.hhmi.org.
  • Hasseman JP; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA. Electronic address: hassemanj@janelia.hhmi.org.
  • Kolb I; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA; GENIE Project Team, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA. Electronic address: kolbi@janelia.hhmi.org.
Neuron ; 111(10): 1547-1563.e9, 2023 05 17.
Article in En | MEDLINE | ID: mdl-37015225
ABSTRACT
The ability to optically image cellular transmembrane voltages at millisecond-timescale resolutions can offer unprecedented insight into the function of living brains in behaving animals. Here, we present a point mutation that increases the sensitivity of Ace2 opsin-based voltage indicators. We use the mutation to develop Voltron2, an improved chemigeneic voltage indicator that has a 65% higher sensitivity to single APs and 3-fold higher sensitivity to subthreshold potentials than Voltron. Voltron2 retained the sub-millisecond kinetics and photostability of its predecessor, although with lower baseline fluorescence. In multiple in vitro and in vivo comparisons with its predecessor across multiple species, we found Voltron2 to be more sensitive to APs and subthreshold fluctuations. Finally, we used Voltron2 to study and evaluate the possible mechanisms of interneuron synchronization in the mouse hippocampus. Overall, we have discovered a generalizable mutation that significantly increases the sensitivity of Ace2 rhodopsin-based sensors, improving their voltage reporting capability.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Rhodopsin / Angiotensin-Converting Enzyme 2 Type of study: Diagnostic_studies Limits: Animals Language: En Journal: Neuron Journal subject: NEUROLOGIA Year: 2023 Type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Rhodopsin / Angiotensin-Converting Enzyme 2 Type of study: Diagnostic_studies Limits: Animals Language: En Journal: Neuron Journal subject: NEUROLOGIA Year: 2023 Type: Article