Light generation of intracellular Ca(2+) signals by a genetically encoded protein BACCS

Nat Commun. 2015 Aug 18:6:8021. doi: 10.1038/ncomms9021.

Abstract

Ca(2+) signals are highly regulated in a spatiotemporal manner in numerous cellular physiological events. Here we report a genetically engineered blue light-activated Ca(2+) channel switch (BACCS), as an optogenetic tool for generating Ca(2+) signals. BACCS opens Ca(2+)-selective ORAI ion channels in response to light. A BACCS variant, dmBACCS2, combined with Drosophila Orai, elevates the Ca(2+) concentration more rapidly, such that Ca(2+) elevation in mammalian cells is observed within 1 s on light exposure. Using BACCSs, we successfully control cellular events including NFAT-mediated gene expression. In the mouse olfactory system, BACCS mediates light-dependent electrophysiological responses. Furthermore, we generate BACCS mutants, which exhibit fast and slow recovery of intracellular Ca(2+). Thus, BACCSs are a useful optogenetic tool for generating temporally various intracellular Ca(2+) signals with a large dynamic range, and will be applicable to both in vitro and in vivo studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Calcium / metabolism
  • Calcium Channels / genetics
  • Calcium Channels / metabolism*
  • Calcium Signaling / physiology*
  • Chlorocebus aethiops
  • Electrophysiological Phenomena
  • Genes, Transgenic, Suicide
  • Genetic Engineering / methods*
  • HEK293 Cells
  • Humans
  • Light*
  • Luminescent Proteins
  • Mice
  • Olfactory Mucosa / metabolism

Substances

  • Calcium Channels
  • Luminescent Proteins
  • Calcium