Polycyclic aromatic hydrocarbons (PAHs) are a contaminant class characterized by fused aromatic rings. PAHs are formed through the incomplete combustion of organic materials and petrogenic sources. Despite the abundance and toxicity of alkyl substituted PAHs, most research and regulation focus on unsubstituted parent PAHs. Alkyl substitution of Benzo[a]pyrene (BaP), one of the most well-studied parent PAHs, drastically alters its bioactivity in zebrafish. In larval zebrafish exposed from 6 hours post-fertilization (hpf), BaP caused behavioral effects but no morphological effects up to 50 µM at 120 hpf. In contrast, 8-methylbenzo[a]pyrene caused a distinct fin duplication phenotype by 0.26 µM and additional morphological effects by 1 µM. Alkyl substitutions in different positions (7-, 6-, 9-, and 10-MBaP) did not elicit morphological effects at similar concentrations. This study characterized the morphological effects of 8-MBaP in zebrafish and investigated its mechanism(s) of action. Using knock-out lines we demonstrated that 8-MBaP toxicity is Ahr2 dependent and that Cyp1a served a protective role. To identify underlying transcriptomic changes, embryos were exposed to three concentrations of BaP, 6-MBaP, and 8-MBaP. Whole embryos/larvae were collected at 48 and 72 hpf which was before and during phenotype onset, respectively. Collecting RNA and morphological effects across concentration, time, and chemicals facilitated the identification of concentration-dependent transcriptional responses linked to the downstream morphological phenotypes unique to BaP methylation at the eight position. This study improves environmental and human health hazard assessment by identifying critical structural features and mechanisms of action contributing to the toxicity of PAH mixtures in the environment.
© The Author(s) 2025. Published by Oxford University Press on behalf of the Society of Toxicology.