Evaluation of atrazine neurodevelopment toxicity in vitro-application of hESC-based neural differentiation model

Atrazine is one of the widely used herbicides in the world and most of the current researches on atrazine neurodevelopment toxicity have focused on rodents or zebrafish models in vivo, resulting in relatively high cost, time consumption, and lower translational value to identify its hazard for the developing brain. Major international initiatives have pushed forward to convert the traditional animal-based developmental toxicity tests to in vitro assays using human cells to detect and predict chemical health hazards. In this study, we presented a human neural differentiation model based on human embryonic stem cells (hESC) that can be used to test toxicity at different stages of neural differentiation in vitro. hESC were differentiated into neural stem cells (NSC) and then terminally differentiated towards mixed neurons and glial cells for 21 days. Cell survival, proliferation, cell cycle, apoptosis, and gene expression levels were examined. Our results demonstrated that atrazine inhibited the proliferation of hESC and NSC, and showed different toxic sensitivity on these two kinds of cells. Also, atrazine blocked the NSC cell cycle G1 phase via down-regulating CCND1, CDK2, and CDK4, with no obvious effect on apoptosis. In addition, atrazine curbed EB spontaneous differentiation and NSC-induced neurons and glia cells differentiation. Atrazine altered genes expression levels of PAX6, TUBB3, NCAM1, GFAP, TH, NR4A1, and GRIA1. From the data we obtained, we recognized that the dopaminergic system was not the only target of atrazine neurotoxicity, glutamatergic neurons and astrocytes were also adversely affected.