Using Transcriptomics and Metabolomics to Understand Species Differences in Sensitivity to Chlorpyrifos in Japanese Quail and Double‐Crested Cormorant Embryos

Abstract

Modern 21st‐century toxicity testing makes use of omics technologies to address critical questions in toxicology and chemical management. Of interest are questions relating to chemical mechanisms of toxicity, differences in species sensitivity, and translation of molecular effects to observable apical endpoints. Our study addressed these questions by comparing apical outcomes and multiple omics responses in early–life stage exposure studies with Japanese quail (Coturnix japonica) and double‐crested cormorant (Phalacrocorax auritus), representing a model and ecological species, respectively. Specifically, we investigated the dose‐dependent response of apical outcomes as well as transcriptomics and metabolomics in the liver of each species exposed to chlorpyrifos, a widely used organophosphate pesticide. Our results revealed a clear pattern of dose‐dependent disruption of gene expression and metabolic profiles in Japanese quail but not double‐crested cormorant at similar chlorpyrifos exposure concentrations. The difference in sensitivity between species was likely due to higher metabolic transformation of chlorpyrifos in Japanese quail compared to double‐crested cormorant. The most impacted biological pathways after chlorpyrifos exposure in Japanese quail included hepatic metabolism, oxidative stress, endocrine disruption (steroid and nonsteroid hormones), and metabolic disease (lipid and fatty acid metabolism). Importantly, we show consistent responses across biological scales, suggesting that significant disruption at the level of gene expression and metabolite profiles leads to observable apical responses at the organism level. Our study demonstrates the utility of evaluating effects at multiple biological levels of organization to understand how modern toxicity testing relates to outcomes of regulatory relevance, while also highlighting important, yet poorly understood, species differences in sensitivity to chemical exposure. Environ Toxicol Chem 2021;40:3019–3033. © 2021 SETAC

Targeted Metabolomics to Assess Exposure to Environmental Chemicals of Concern in Japanese Quail at Two Life Stages

“…Overall, few effects of chemical exposure on the metabolome were observed at the doses that we tested. This was notable, given that significant changes were reported at the transcriptional and organismal level in response to several of the chemicals [ 11 , 13 ]. Here, we discuss these results and factors that may have contributed to this outcome.…”

“…Among them, 16 lipids showed a log2FC > |1.5| ( Table 1 ). We previously explored this result in a related study comparing transcriptomic and metabolomic responses to CPF in two avian species: Japanese quail and double-crested cormorant ( Phalacrocorax auritus ) [ 11 ]. We found that CPF had a high impact in Japanese quail ELS liver, disrupting hepatic metabolism, causing oxidative stress, and endocrine disruption (steroid and non-steroid hormones).…”

“…toxin, respectively [8,9]. Recently, a metabolomics study in Japanese quail embryos revealed effects on lipid and fatty-acid metabolism following chlorpyrifos exposure [11].…”

“…To obtain sufficient wet weight required for metabolomics analysis, livers from three to four individuals were pooled, constituting one replicate, and three replicates were collected per dose. Additional liver samples were collected for other analyses, such as analytical determination of the concentrations of test chemicals in tissue, transcriptomics, proteomics, and histology [ 11 , 13 ]. Tissue samples were flash frozen on dry ice and stored at −80 °C until further analysis.…”

“…In addition, amino acid, energy, fatty acid, and nucleoside metabolisms were disrupted in double-crested cormorant and chicken after exposure to Deep Water Horizon oil and T2-toxin, respectively [8,9]. Recently, a metabolomics study in Japanese quail embryos revealed effects on lipid and fatty-acid metabolism following chlorpyrifos exposure [11]. Here, we describe a proof-of-concept metabolomic study in the Japanese quail (Coturnix japonica) after exposure to anthropogenic contaminants at two life stages.…”