Tri-iso-butyl phosphate (TiBP) exposure induces neurotoxicity by triggering oxidative stress accompanied by neurotransmitter system disruptions and apoptosis in zebrafish larvae

The current research sheds light on the biological toxicity of organophosphate flame retardants (OPFRs), yet it overlooks the neurotoxicity and potential molecular mechanisms of tris(1,3-dichloro-2-propyl) phosphate (TiBP), a prominent constituent of the OPFRs. To address this, we utilized zebrafish larvae as a model to investigate TiBP's acute toxicity and neurotoxic effects, along with the associated molecular pathways. Our findings revealed that the 96h and 120h LC50 values for TiBP were 56.51 mg/L and 48.85 mg/L, respectively. Gradient exposure based on the 120h LC50 demonstrated that TiBP induced developmental toxicity, characterized by elevated heart rate, reduced body length, and diminished eye distance. Additionally, a decrease in swimming activity was observed in the light test, along with the inhibition of the neuro crest cell development in Tg (HuC:eGFP) and Tg (sox10: eGFP) zebrafish larvae following TiBP exposure, as well as the alterations of neurogenesis and ACh-related genes. Expression of key neurodevelopment genes, including mbpa, gap43, nestin, ngfra, was significantly downregulated. Furthermore, heightened anxiety-like behaviors in open field and phototaxis tests were observed, concomitant with neurotransmitter imbalances. Specifically, there was an increase in DA levels, a decrease in GABA, and an upregulation of AChE activity. These disruptions were primarily mediated through transcriptional dysregulation of neurotransmitter synthesis, transport, and reception. Upon exposure to TiBP, zebrafish larvae exhibited a concentration-dependent increase in both ROS level and apoptosis. An upregulation of antioxidant enzymes and their transcription levels indicated the presence of oxidative stress in the larvae. The induction of ddit3 was congruent with the observed apoptosis, suggesting that it may be triggered by oxidative stress via the ERs-CHOP pathway. In summary, our study indicates that oxidative stress is a pivotal molecular event in the neurotoxicity induced by TiBP, implicating the disruption of the GABAergic, dopaminergic, and cholinergic systems, as well as triggering apoptosis.