The neurotoxicity of Chlorantraniliprole to Helicoverpa armigera larvae.

Chlorantraniliprole (CAP) is a mainstream diamide insecticide widely used against lepidopteran pests such as Helicoverpa armigera. Given that most insecticides act via neurotoxic mechanisms, we reasoned that CAP, despite its well-established action on ryanodine receptors (RyRs), may also disrupt nervous system function through additional targets. Insect neurotoxicity is a coordinated process involving neurons and glial cells: voltage-gated sodium (Nav) and potassium (Kv) channels are common insecticide targets, however, their involvement in CAP-induced neurotoxicity has not been established, and glial contributions are poorly understood. We elucidated CAP's multi-target mechanisms by constructing the first single-cell transcriptomic atlas of the H. armigera ventral nerve cord (VNC) using scRNA-seq, validated by whole-cell patch-clamp electrophysiology and RT-qPCR. Results showed that CAP modulated the kinetics of Nav channel in a concentration-dependent manner and suppressed Kv channel currents, confirming both as novel non-canonical targets. scRNA-seq identified VNC cell types, identifying NKCC (perineurial glia-enriched) and UGT39B1 (subperineurial glia-enriched); both were upregulated after 24 h LC30 CAP exposure. These results indicate that CAP modulates Nav and Kv channels at relatively high concentrations. As potential functional targets, these channels supplement the primary RyR-mediated action of CAP, and imply a partial mechanistic similarity to pyrethroids via regulation of Nav channels. Moreover, the VNC atlas facilitated the identification of glial-specific markers (NKCC and UGT39B1), clarifying glial involvement. Collectively, our work advances multi-target understanding of diamide insecticides, provides candidate synergists, and supports "neuron-glial dual-target" strategies for managing H. armigera resistance.