Thiophanate-methyl induces notochord toxicity by activating the PI3K-mTOR pathway in zebrafish (Danio rerio) embryos

Thiophanate-methyl (TM), a typical pesticide widely used worldwide, was detected in rivers, soil, fruits, and vegetables. Thus, it is urgent to identify the potential harm of TM residual to non-target organisms and its molecular mechanisms. We used zebrafish (Danio rerio) in this study to evaluate TM toxicity. TM exposure induced developmental toxicity, including inhibited hatchability, reduced heart rates, restrained spontaneous locomotion, and decreased body length. Furthermore, we observed obvious toxicity in the notochord and detected increased expression levels of notochord-related genes (shha, col2a, and tbxta) by in situ hybridization in zebrafish larvae. In addition, calcein staining, alkaline phosphatase (ALP) activity analysis, and anatomic analysis indicated that TM induced notochord toxicity. We used rescue experiments to verify whether the PI3K-mTOR pathway involved in the notochord development was the cause of notochord abnormalities. Rapamycin and LY294002 (an inhibitor of PI3K) relieve notochord toxicity caused by TM, including morphological abnormalities. In summary, TM might induce notochord toxicity by activating the PI3K-mTOR pathway in zebrafish.