Tracking AC Electric Stimulation-Induced Persistent Locomotion Behavior in the Nematode Caenorhabditis elegans

Persistent neural activity underlies fundamental brain functions such as memory, decision-making, and emotion. Despite its importance, experimental paradigms that enable quantitative analysis of persistent behavioral responses remain limited. Here, we describe a protocol to induce and measure a persistent locomotor response by applying a brief alternating current (AC) electric stimulus to the nematode Caenorhabditis elegans. This method reliably evokes a prolonged increase in locomotion speed that persists for minutes after stimulus termination and can be quantified by video tracking. Because C. elegans has a fully mapped connectome and is amenable to genetic and neurophysiological manipulation, this protocol provides a useful platform for dissecting the molecular and neural mechanisms underlying persistent behavioral responses. Electrically induced persistent locomotion serves as a simple, robust, and quantifiable behavioral readout for studying the regulation of neural persistence in vivo. Key features • A brief AC electric stimulus reliably induces a persistent locomotor response in C. elegans. • The assay uses a simple setup and provides reproducible behavioral data. • Video-tracking analysis enables quantitative measurement of locomotion speed and response persistence. • This protocol enables quantitative comparisons and analyses of behavioral changes evoked by stimulus characteristics and/or genetic mutations.