Single-cell transcriptomics unveils atrazine's impact on neurons and microglia in C57BL/6 mice

BACKGROUND: Atrazine (ATZ), a widely used herbicide, is implicated in neurodegenerative risks, yet its neurotoxic mechanisms remain unclear. This study investigates how environmentally relevant ATZ exposure disrupts neuron-microglia interactions to drive Parkinson's disease (PD)-like pathology. METHODS: C57BL/6 mice received 28-day oral ATZ (10 mg/kg/day). Behavioral phenotyping (open field, pole climb, wire hanging tests) assessed motor deficits. Midbrain tissues underwent histopathology and single-cell RNA sequencing (scRNA-seq). Intercellular communication networks were reconstructed using the CellChat algorithm, with a focus on neuron-microglia signaling pathways. Quantitative real-time PCR (qPCR) was employed to validate the transcriptomic accuracy of scRNA-seq (n = 6/group). RESULT: ATZ induced PD-like motor dysfunction (e.g., mean speed in OFT, P < 0.0001) and neuronal damage. scRNA-seq identified dysregulated calcium homeostasis genes (Atp2b1, Camk2a, Gabbr2) and neurotoxic markers (Mapt, St18) in neurons, alongside M1 microglia polarization via TLR/TNF/IL-17 signaling. Microglial-derived CX3CL1 was found to competitively disrupt neuronal CX3CL1-CX3CR1 signaling, exacerbating neuroinflammation. qPCR indicated high accuracy of the scRNA-seq data. CONCLUSION: Environmental ATZ exposure triggers PD-like neurodegeneration through dual mechanisms: (1) neuronal calcium dysregulation inducing oxidative stress and (2) microglia-driven neuroimmune dysfunction via CX3CL1 signaling. This study provides the novel mechanistic evidence linking ATZ to PD-like pathology via neuron-microglia crosstalk, highlighting the need for re-evaluating global ATZ exposure guidelines.