A comparison of the effects of ivermectin and moxidectin on the nematode Caenorhabditis elegans

Abstract

The avermectins and the milbemycins are structurally related classes of 16-membered macrocyclic lactones (ML) that have a broad spectrum of activity. Most studies on the mode of action of ML have used the avermectin, ivermectin (IVM). IVM activates glutamate-gated chloride channels that contain alpha-type subunits, resulting in a hyperpolarization of the neuronal membrane, leading to a flaccid paralysis. IVM kills Caenorhabditis elegans at therapeutic concentrations, making it a useful model to examine mechanisms of IVM toxicity and resistance. There have been suggestions that the milbemycins may exert effects that are different from the avermectins, however this hypothesis has been challenged. Using IVM and the milbemycin, moxidectin (MOX), we demonstrate that while the two drugs have some similar effects on C. elegans, there are also some differences in worm response. Following exogenous exposure to a gradient of IVM and MOX, ranging from 0 to 5000 nM, quantitative and qualitative differences in response to the two anthelmintic drugs were observed in the pharyngeal pump rate, larval development and motility of wild-type and glutamate-gated chloride channel (GluCl) subunit knockout strains of C. elegans. After exposure to equimolar drug concentrations, differences between the anthelmintic effects were observed in the motility phenotype in the wild-type, GluCl subunit knockout strains and multi-gene knockout strain of C. elegans that exhibits a marked reduction in IVM sensitivity; and transcription profiles of genes coding for GluCl subunits in both the wild-type and glc-2 knockout strain. The glc-2 deletion strain showed increased motility in response to 2.5 nM MOX in the first 1.5 h of exposure, compared with wild-type nematodes, whereas this strain showed little change in motility in response to IVM. The pharyngeal pump rate in the glc-2 deletion strain was sensitive to equimolar concentrations of IVM and MOX. The triple avr-14/ avr-15/ glc-1 knockout caused a loss of initial stimulation of motility seen in the wild-type, by 2.5 nM IVM, to a reduction in motility, whereas the response to MOX was little changed between this triple knockout strain and wild-type C. elegans. The results suggest that there are significant differences in the response of C. elegans to IVM and MOX. The product of the glc- 2 gene may play a role in sensitivity to MOX, but not to IVM, while the products of avr-14, avr-15 and glc-1 may be important for the effects of IVM, but less so for MOX.