Archaerhodopsin variants with enhanced voltage-sensitive fluorescence in mammalian and Caenorhabditis elegans neurons

Nat Commun. 2014 Sep 15:5:4894. doi: 10.1038/ncomms5894.

Abstract

Probing the neural circuit dynamics underlying behaviour would benefit greatly from improved genetically encoded voltage indicators. The proton pump Archaerhodopsin-3 (Arch), an optogenetic tool commonly used for neuronal inhibition, has been shown to emit voltage-sensitive fluorescence. Here we report two Arch variants with enhanced radiance (Archers) that in response to 655 nm light have 3-5 times increased fluorescence and 55-99 times reduced photocurrents compared with Arch WT. The most fluorescent variant, Archer1, has 25-40% fluorescence change in response to action potentials while using 9 times lower light intensity compared with other Arch-based voltage sensors. Archer1 is capable of wavelength-specific functionality as a voltage sensor under red light and as an inhibitory actuator under green light. As a proof-of-concept for the application of Arch-based sensors in vivo, we show fluorescence voltage sensing in behaving Caenorhabditis elegans. Archer1's characteristics contribute to the goal of all-optical detection and modulation of activity in neuronal networks in vivo.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / physiology*
  • Animals
  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Caenorhabditis elegans / chemistry
  • Caenorhabditis elegans / metabolism
  • Gene Expression
  • Helminth Proteins / chemistry*
  • Helminth Proteins / genetics
  • Hippocampus / chemistry
  • Hippocampus / metabolism
  • Light
  • Nerve Tissue Proteins / chemistry*
  • Nerve Tissue Proteins / genetics
  • Neurons / chemistry*
  • Neurons / metabolism
  • Optogenetics / methods
  • Patch-Clamp Techniques
  • Primary Cell Culture
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Rats
  • Rats, Wistar
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Spectrometry, Fluorescence

Substances

  • Archaeal Proteins
  • Helminth Proteins
  • Nerve Tissue Proteins
  • Protein Isoforms
  • Recombinant Proteins
  • archaerhodopsin protein, Archaea