Channelrhodopsin-2-XXL, a powerful optogenetic tool for low-light applications

Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):13972-7. doi: 10.1073/pnas.1408269111. Epub 2014 Sep 8.

Abstract

Channelrhodopsin-2 (ChR2) has provided a breakthrough for the optogenetic control of neuronal activity. In adult Drosophila melanogaster, however, its applications are severely constrained. This limitation in a powerful model system has curtailed unfolding the full potential of ChR2 for behavioral neuroscience. Here, we describe the D156C mutant, termed ChR2-XXL (extra high expression and long open state), which displays increased expression, improved subcellular localization, elevated retinal affinity, an extended open-state lifetime, and photocurrent amplitudes greatly exceeding those of all heretofore published ChR variants. As a result, neuronal activity could be efficiently evoked with ambient light and even without retinal supplementation. We validated the benefits of the variant in intact flies by eliciting simple and complex behaviors. We demonstrate efficient and prolonged photostimulation of monosynaptic transmission at the neuromuscular junction and reliable activation of a gustatory reflex pathway. Innate male courtship was triggered in male and female flies, and olfactory memories were written through light-induced associative training.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Evoked Potentials, Visual*
  • Female
  • Male
  • Mutation, Missense*
  • Neurons / metabolism*
  • Rhodopsin / genetics
  • Rhodopsin / metabolism*
  • Synaptic Transmission*

Substances

  • Rhodopsin