Anxiety caused by chronic exposure to methylisothiazolinone in zebrafish: behavioral analysis, brain histology and gene responses.

Abstract

Methylisothiazolinones (MIT) are a class of preservatives and biocides extensively utilized in everyday products, industrial processes, and medical and healthcare applications. However, reports have indicated that MIT may cause skin irritation and neurotoxicity. Given its pervasive use, the neurotoxic potential of MIT has garnered increasing attention. Recent in vitro cellular experiments have demonstrated that MIT inhibits synaptic growth, although the neurotoxic effects and underlying mechanisms at the organismal level remain largely unexplored. In this study, it was found for the first time that long-term exposure to MIT resulted in anxiety, brain tissue inflammation, and a reduction in the number of Nissl bodies in the brain. Additionally, transcriptomic analysis indicated that exposure to 300 μg/L MIT induced a greater number of differentially expressed genes compared to 30 μg/L MIT, relative to the control group. Enrichment analysis, trend analysis, and GSEA analysis collectively identified the involvement of Steroid hormone metabolism, oxidative metabolism, and the Hedgehog pathway in MIT-induced neurotoxicity. Furthermore, a subsequent reduction in green fluorescence was observed in the MLS-EGFP zebrafish strain larvae of the HD group, suggesting that high dosage of MIT exerts an inhibitory effect on mitochondrial activity. This study confirmed the neurotoxic effects of MIT and investigated the potential genetic networks behind anxiety behavior. These findings contributed to the identification of key brain genes involved in the detection and monitoring of MIT, offering new insights into the neuroendocrine toxicity of other imidazolidinone compounds.