Proteomic profiling of valproic acid-treated zebrafish embryos highlights dysregulation in energy and purine metabolism, and microtubule dynamics: Implications for autism spectrum disorder.

Valproate (VPA), a widely used anticonvulsant, is also employed to establish experimental autism spectrum disorder (ASD) models. This study aims to elucidate mechanisms underlying VPA's effects in ASD by analyzing proteomic profiles and oxidant-antioxidant dynamics in zebrafish embryos, uncovering the cellular pathways driving these changes. Zebrafish embryos were exposed to two concentrations of VPA (10 μM and 25 μM) for 72 h post-fertilization (hpf), and LC-MS/MS analyses were performed. Differentially expressed proteins (DEPs) were subjected to bioinformatic analysis to identify associated cellular pathways, and their biological significance was evaluated. Oxidant-antioxidant parameters and locomotor activities were determined. High-dose induced more pronounced proteomic changes, while most of the identified proteins in both groups, including key metabolic enzymes such as adenylate kinase 1 (Ak1), adenosine monophosphate deaminase 1 (Ampd1), pyruvate kinase (Pkmb) and creatine kinase (Ck), exhibited a dose-dependent decrease. Functional enrichment analyses revealed that these alterations were primarily associated with purine metabolism, energy metabolism, and microtubule dynamics. In addition, malondialdehyde, nitric oxide, and glutathione S-transferase, increased in a dose-dependent manner, whereas superoxide dismutase decreased. Decreased average speed, distance swam, and explored areas were found in both VPA treated groups, reflecting early sensorimotor dysfunction. Our findings demonstrate that VPA induces dose-dependent proteomic alterations in zebrafish embryos, with metabolic pathways and cytoskeletal dynamics being particularly affected. Extent of molecular disruptions appears to correlate with VPA concentration, suggesting potential implications for energy homeostasis and cellular structure. Understanding these effects could provide valuable insights into the developmental toxicity mechanisms of VPA and its broader biological significance.