Neurotoxicity Evaluation of Nanomaterials Using C. elegans: Survival, Locomotion Behaviors, and Oxidative Stress

Nanomaterials are broadly used in a variety of industries and consumer products. However, studies have demonstrated that many nanomaterials, including metal-containing nanoparticles and nanoplastics, have neurotoxic effects. Caenorhabditis elegans (C. elegans) is a widely used model organism with numerous advantages for research, including transparency, short life span, well-characterized nervous system, complete connectome, available genome, and numerous genetic tools. C. elegans has been extensively used to assess the neurotoxicity of multiple chemicals via survival assays, behavioral tests, neuronal morphology studies, and various molecular and mechanistic analyses. However, detailed protocols describing general assays in C. elegans to examine the neurotoxic effects of nanomaterials are limited. Here, we describe protocols for assessing nanomaterial neurotoxicity in C. elegans. We describe the steps for exposure and subsequent evaluation of survival, locomotion behavior, and oxidative stress. Survival and locomotion behavior are measured in wild-type N2 strains to assess acute neurotoxicity. Oxidative stress is used as an endpoint here since it is one of the most predominant and common changes induced by nanomaterials. VP596 nematodes, which express GFP upon activation of skn-1 (the worm homolog of Nrf2), are evaluated for assays of oxidative stress in response to test nanomaterials. These assays can be readily used to quickly examine the neurotoxicity of nanomaterials in vivo, laying the foundation for mechanistic studies of nanomaterials and their impacts on health and physiology. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Exposure of C. elegans to nanomaterials Basic Protocol 2: Survival assessment Basic Protocol 3: Assessment of locomotion behavior Basic Protocol 4: Analysis of oxidative stress.