Silencing neurotransmission with membrane-tethered toxins.
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.
Molecular Neurobiology group, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
Sebastian
S
Auer
Annika S
AS
Stürzebecher
René
R
Jüttner
Julio
J
Santos-Torres
Christina
C
Hanack
Silke
S
Frahm
Beate
B
Liehl
Inés
I
Ibañez-Tallon
D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D020864:Calcium Channels, N-Type / Q000187:drug effects; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism; D006801:Humans; D019426:Integrases / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010146:Pain / Q000517:prevention & control; D051381:Rats; D017208:Rats, Wistar; D009435:Synaptic Transmission / Q000187:drug effects*; D020960:omega-Conotoxins / Q000378:metabolism / Q000494:pharmacology*
D002121:Calcium Channel Blockers; D020864:Calcium Channels, N-Type; D004298:Dopamine; D019426:Integrases; D020960:omega-Conotoxins; C500545:CACNA1B protein, human; C078452:ziconotide; C045073:Cre recombinase