A Photoactivatable Botulinum Neurotoxin for Inducible Control of Neurotransmission

Neuron. 2019 Mar 6;101(5):863-875.e6. doi: 10.1016/j.neuron.2019.01.002. Epub 2019 Jan 28.

Abstract

Regulated secretion is critical for diverse biological processes ranging from immune and endocrine signaling to synaptic transmission. Botulinum and tetanus neurotoxins, which specifically proteolyze vesicle fusion proteins involved in regulated secretion, have been widely used as experimental tools to block these processes. Genetic expression of these toxins in the nervous system has been a powerful approach for disrupting neurotransmitter release within defined circuitry, but their current utility in the brain and elsewhere remains limited by lack of spatial and temporal control. Here we engineered botulinum neurotoxin B so that it can be activated with blue light. We demonstrate the utility of this approach for inducibly disrupting excitatory neurotransmission, providing a first-in-class optogenetic tool for persistent, light-triggered synaptic inhibition. In addition to blocking neurotransmitter release, this approach will have broad utility for conditionally disrupting regulated secretion of diverse bioactive molecules, including neuropeptides, neuromodulators, hormones, and immune molecules. VIDEO ABSTRACT.

Keywords: Optogenetics; SNARE protein; botulinum toxin; iLID; neurotransmitter; secretion; synapse; tetanus toxin; vamp2; vesicle.

Publication types

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

MeSH terms

  • Animals
  • Arabidopsis Proteins / genetics
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Botulinum Toxins / genetics
  • Botulinum Toxins / pharmacology*
  • Botulinum Toxins / radiation effects
  • Caenorhabditis elegans
  • Cells, Cultured
  • Cryptochromes / genetics
  • Female
  • HEK293 Cells
  • Humans
  • Light
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Optogenetics / methods*
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacology
  • Recombinant Proteins / radiation effects
  • SNARE Proteins / metabolism
  • Synapses / metabolism
  • Synapses / physiology
  • Synaptic Transmission / drug effects*
  • Vesicle-Associated Membrane Protein 2 / metabolism

Substances

  • Arabidopsis Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • CIB1 protein, Arabidopsis
  • CRY2 protein, Arabidopsis
  • Cryptochromes
  • Recombinant Proteins
  • SNARE Proteins
  • Vesicle-Associated Membrane Protein 2
  • Botulinum Toxins