Dorsal Unpaired Median Neurons ofLocusta migratoriaExpress Ivermectin- and Fipronil-Sensitive Glutamate-Gated Chloride Channels

Abstract

Together with type A GABA and strychnine-sensitive glycine receptors, glutamate-gated chloride channels (GluCl) are members of the Cys-loop family of ionotropic receptors, which mediate fast inhibitory neurotransmission. To date, GluCls are found in invertebrates only and therefore represent potential specific targets for insecticides, such as ivermectin and fipronil. In this study, we identified the functional expression of GluCls in dorsal unpaired median (DUM) neurons of the metathoracic ganglion of Locusta migratoria using electrophysiological and molecular biological techniques. In whole cell patch-clamped DUM neurons, glutamate-induced changes in both their membrane potentials (current-clamp) and currents (voltage-clamp) were dependent on the chloride equilibrium potential. On continuous application of glutamate, the glutamate-elicited current response became rapidly and completely desensitized. Application of glutamate in the presence of 10 μM fipronil or 100 μM picrotoxin reversibly decreased GluCl-mediated currents by 87 and 39%, respectively. Furthermore, 1 μM ivermectin induced a persistent chloride current, suggesting the expression of ivermectin-sensitive GluCl α subunits. A degenerate PCR/RACE strategy was used to clone the full-length L. migratoria LmGlClα subunit. Finally, RT-PCR experiments demonstrated the presence of LmGluClα transcripts in locust DUM neurons. Our results provide the first direct evidence of a functional ivermectin-sensitive GluCl channel on the cell surface of DUM neurons of L. migratoria.