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Published September 1, 2002 | Published
Journal Article Open

Selective Electrical Silencing of Mammalian Neurons In Vitro by the Use of Invertebrate Ligand-Gated Chloride Channels


Selectively reducing the excitability of specific neurons will (1) allow for the creation of animal models of human neurological disorders and (2) provide insight into the global function of specific sets of neurons. We focus on a combined genetic and pharmacological approach to silence neurons electrically. We express invertebrate ivermectin (IVM)-sensitive chloride channels (Caenorhabditis elegans GluCl α and β) with a Sindbis virus and then activate these channels with IVM to produce inhibition via a Cl^− conductance. We constructed a three-cistron Sindbis virus that expresses the α and β subunits of a glutamate-gated chloride channel (GluCl) along with the green fluorescent protein (EGFP) marker. Expression of the C. elegans channel does not affect the normal spike activity or GABA/glutamate postsynaptic currents of cultured embryonic day 18 hippocampal neurons. At concentrations as low as 5 nM, IVM activates a Cl^− current large enough to silence infected neurons effectively. This conductance reverses in 8 hr. These low concentrations of IVM do not potentiate GABA responses. Comparable results are observed with plasmid transfection of yellow fluorescent protein-tagged (EYFP) GluCl α and cyan fluorescent protein-tagged (ECFP) GluCl β. The present study provides an in vitromodel mimicking conditions that can be obtained in transgenic mice and in viral-mediated gene therapy. These experiments demonstrate the feasibility of using invertebrate ligand-activated Cl^− channels as an approach to modulate excitability.

Additional Information

© 2002 Society for Neuroscience. Received March 25, 2002; Revised June 5, 2002; Accepted June 13, 2002. This work was supported by National Institutes of Health Grants NS 11756 and MH 49176, by the Sidney Stern and Plum Foundations, and by the William T. Gimbal Discovery fund in Neuroscience. We thank Doris Cully for generously supplying the C. elegans GluCl channel cDNAs and for discussion; Birgit Hirschberg, Charles Cohen, and Christof Koch for discussion; and Dong Ju for technical assistance.

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