Genetic Evidence for Estrogenic Effects of Benzophenone-2 on Zebrafish Neurodevelopment and Its Signaling Mechanism

Estrogens play a crucial role in regulating various biological responses during the early stages of neurodevelopment. Benzophenone-2 (BP2), a widely used organic ultraviolet (UV) filter, has been proven as an estrogenic compound, whereas the estrogenic effects of BP2 on neurodevelopment remain largely unknown. Here, we investigated the neurodevelopmental toxicity of BP2 by exposing zebrafish embryos from 2 to 120 h postfertilization (hpf) at environmentally relevant concentrations. We demonstrated that early life exposure to BP2 induced multiple concentration-dependent impairments in the nervous system, including hypoactivity, abnormal brain morphology, impaired neurocyte proliferation, shortened axon, and increased neurocyte apoptosis. Moreover, metabolomic profiling revealed a decrease in dopamine (DA) and its metabolites in BP2-treated larvae. Using E2 treatment and morpholino knockdown assays, we provided strong genetic evidence that the BP2-induced behavioral disorders were associated with estrogen-dependent signaling, especially estrogen receptors 2a and 2b (<i>esr2</i>). Subsequently, transcriptomic profiling indicated that the activation of <i>esr2</i> further inhibited the expression of LIM homeobox transcription factor 1 β a (<i>lmx1ba)</i>, which is vital for normal neurodevelopment. Consistently, the overexpression of <i>lmx1ba</i> and inhibition of <i>esr2</i> obviously alleviated BP2-caused neurotoxicity, uncovering a seminal role of <i>esr2</i> and <i>lmx1ba</i> in BP2-induced neurodevelopmental toxicity. Our findings provide the first evidence in fish that BP2 can induce neurodevelopmental deficits and brain dysfunction and offer novel insights into the mechanisms of toxicity of BP2 as well as other emerging benzophenones.