Silencing neurotransmission with membrane-tethered toxins

Nat Methods. 2010 Mar;7(3):229-36. doi: 10.1038/nmeth.1425. Epub 2010 Feb 7.


At synaptic terminals, high voltage-activated Ca(v)2.1 and Ca(v)2.2 calcium channels have an essential and joint role in coupling the presynaptic action potential to neurotransmitter release. Here we show that membrane-tethered toxins allowed cell-autonomous blockade of each channel individually or simultaneously in mouse neurons in vivo. We report optimized constitutive, inducible and Cre recombinase-dependent lentiviral vectors encoding fluorescent recombinant toxins, and we also validated the toxin-based strategy in a transgenic mouse model. Toxins delivered by lentiviral vectors selectively inhibited the dopaminergic nigrostriatal pathway, and transgenic mice with targeted expression in nociceptive peripheral neurons displayed long-lasting suppression of chronic pain. Optimized tethered toxins are tools for cell-specific and temporal manipulation of ion channel-mediated activities in vivo, including blockade of neurotransmitter release.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium Channel Blockers / pharmacology*
  • Calcium Channels, N-Type / drug effects
  • Cells, Cultured
  • Dopamine / metabolism
  • Humans
  • Integrases / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Pain / prevention & control
  • Rats
  • Rats, Wistar
  • Synaptic Transmission / drug effects*
  • omega-Conotoxins / metabolism
  • omega-Conotoxins / pharmacology*


  • CACNA1B protein, human
  • Calcium Channel Blockers
  • Calcium Channels, N-Type
  • omega-Conotoxins
  • voltage-dependent calcium channel (P-Q type)
  • ziconotide
  • Cre recombinase
  • Integrases
  • Dopamine