Integrated multi-omics analysis identifies PI3K-Akt signaling as a candidate pathway in glufosinate-induced neurotoxicity.

Glufosinate-ammonium (GLA), a widely used phosphinic acid herbicide, has raised concerns regarding its potential neurotoxic effects. Although acute GLA exposure frequently induces seizures, the underlying molecular responses remain incompletely understood. In this study, we integrated network toxicology, hippocampal transcriptomics, targeted neurochemical analysis, and experimental validation to characterize the molecular responses associated with acute GLA exposure. Acute GLA exposure induced seizures accompanied by enhanced hippocampal spike activity and neuronal activation. Network analysis implicated amino acid metabolism in the molecular response to GLA exposure, whereas integrated transcriptomic analysis identified molecular responses associated with neuronal survival and cell fate. Integrated pathway analysis further prioritized PI3K-Akt signaling pathway as a candidate pathway for experimental evaluation. These predicted molecular responses and signaling alterations were subsequently evaluated experimentally. Targeted neurochemical analysis revealed only subtle alterations in amino acid metabolism and did not provide evidence supporting glutamate-mediated excitotoxicity. Quantitative PCR confirmed selected transcriptional changes related to cellular remodeling, while increased AKT phosphorylation together with a modest prolongation of the restricted mean seizure-free time following LY294002 treatment provided supportive evidence for the potential involvement of PI3K-Akt signaling. Collectively, this study provides a systems-level characterization of molecular responses to acute GLA exposure and highlights PI3K-Akt signaling as a candidate pathway for future mechanistic investigation.