Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging

Sci Rep. 2017 Feb 13:7:42398. doi: 10.1038/srep42398.

Abstract

We report development of the first genetically encoded bioluminescent indicator for membrane voltage called LOTUS-V. Since it is bioluminescent, imaging LOTUS-V does not require external light illumination. This allows bidirectional optogenetic control of cellular activity triggered by Channelrhodopsin2 and Halorhodopsin during voltage imaging. The other advantage of LOTUS-V is the robustness of a signal-to-background ratio (SBR) wherever it expressed, even in the specimens where autofluorescence from environment severely interferes fluorescence imaging. Through imaging of moving cardiomyocyte aggregates, we demonstrated the advantages of LOTUS-V in long-term imaging are attributable to the absence of phototoxicity, and photobleaching in bioluminescent imaging, combined with the ratiometric aspect of LOTUS-V design. Collectively LOTUS-V extends the scope of excitable cell control and simultaneous voltage phenotyping, which should enable applications in bioscience, medicine and pharmacology previously not possible.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cells, Cultured
  • Electrophysiological Phenomena
  • Gene Expression*
  • Genes, Reporter*
  • Humans
  • Induced Pluripotent Stem Cells / metabolism
  • Kinetics
  • Luminescent Measurements* / methods
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism*
  • Models, Molecular
  • Molecular Imaging* / methods
  • Optogenetics* / methods
  • Protein Conformation

Substances

  • Luminescent Proteins