Spatially resolved hepatic lipid metabolism perturbation and gut microbiota dysbiosis induced by a novel PFOA alternative in zebrafish (Danio rerio).

Abstract

The distribution of environmental pollutants and their associated metabolic perturbations within tissues is essential for understanding toxicological mechanisms. However, whether hexafluoropropylene oxide tetramer acid (HFPO-TeA), an emerging alternative to legacy PFASs, induces spatially heterogeneous metabolic disruption in aquatic organisms remains unclear. Here, adult zebrafish were exposed to HFPO-TeA to investigate hepatic phospholipid spatial distribution and gut microbial dysfunction. Matrix-assisted laser desorption/ionization quadrupole time-of-flight mass spectrometry imaging (MALDI-QTOF-MSI) revealed marked spatial perturbation of 14 putatively annotated lipid features, mainly including phosphatidylcholines (PCs), phosphatidylglycerols (PGs), phosphatidylethanolamines (PEs), diacylglycerols (DAGs), and phosphatidylinositols (PIs). Notably, these phospholipids exhibited region-specific down- or up-regulation, suggesting localized phospholipid remodeling within hepatic microdomains rather than spatially uniform disruption. These spatially resolved phospholipid alterations provide mechanistically relevant evidence for HFPO-TeA-induced hepatotoxicity and phospholipids with annotated spatial perturbation as candidate lipid-feature biomarkers for future validation. Moreover, HFPO-TeA reduced gut microbial diversity and reshaped the intestinal community, including decreases in Firmicutes at the phylum level and increases in Shewanella and Aeromonas at the genus level, taxa linked to lipid metabolism and phospholipid remodeling. Functional prediction further indicated suppressed microbial metabolic potential, with relative enrichment of genetic information processing and cellular maintenance functions. Together, the microbiome shift toward phospholipid remodeling-associated taxa and the spatially resolved hepatic perturbation of specific phospholipids provide convergent evidence that targeted phospholipid metabolic remodeling within tissue microregions may underlie HFPO-TeA-induced metabolic toxicity. This study provides a spatially resolved perspective on the multilevel metabolic toxicity of HFPO-TeA and demonstrates significant potential for revealing the health impacts of emerging PFAS alternatives.