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Phenanthrene is a low molecular weight polycyclic aromatic hydrocarbon (PAH) that is composed of three fused benzene rings. PAHs are formed naturally through incomplete combustion of organic materials, and are environmental contaminants due to anthropogenic activities (e.g. oil extraction and refining, industrial and municipal effluents, fossil fuel burning). Fish exposed to PAHs such as phenanthrene have been reported to exhibit altered reproductive axis endpoints, however the mechanisms that underlie these responses are not fully characterized. To better understand effects at the mechanistic level, we applied transcriptomics to identify molecular pathways altered after acute exposure to phenanthrene on both a dose and temporal scale. Female fathead minnow (Pimephales promelas) were exposed to an average measured concentration of either 0, 29.8, 389 or 943 μg phenanthrene/L for 24, 48, and 72 h in a static-renewal bioassay. Ovaries were assessed for oocyte distribution as well as in vitro 17β-estradiol production and gene expression for transcripts related to steroidogenesis and estrogen signalling. In addition, the liver transcriptome was measured as this tissue is the primary source of the egg yolk precursor protein vitellogenin. Exposure to 29.8 μg phenanthrene/L increased proportions of the cortical alveolar stage in the ovaries after 48 h while the proportion of cortical alveolar oocyte were decreased in fish exposed to 943 μg phenanthrene/L for 48 h. Phenanthrene did not affect 17β-estradiol production at any time or dose, and did not affect transcripts associated with hormone synthesis nor signalling pathways. In the liver, the transcriptome showed fewer genes in common across time when compared to those transcripts affected by concentration at a single time point. Cholesterol metabolism was the only pathway perturbed in the liver following all comparisons in both the dose and time course experiments. Our data suggest that transcriptome networks associated with hepatic lipid metabolism are rapidly affected by phenanthrene, and this may indirectly reduce resources available for reproductive efforts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Phenanthrene is a tricyclic polycyclic aromatic hydrocarbon and environmental contaminant found in high concentrations around urban catchments and in the vicinity of oil extraction activities. Fish exposed to phenanthrene can exhibit altered reproductive hormone profiles and/or differences within gonadosomatic index and altered gamete proportions, but the mechanisms underlying these changes are not fully understood. In this study, we conducted a sub-chronic bioassay and measured transcriptional responses in the liver, the major tissue involved in generating lipids for oocyte growth. Adult male and female fathead minnow (Pimephales promelas) were exposed to an average measured concentration of 202 μg phenanthrene/L for a 7 week period. Condition factor was reduced in both males and females, while female fish also showed decreased gonadosomatic index relative to control females. In females exposed to phenanthrene, perinucleolar proportions were increased ∼1.9-fold relative to the control group whereas the proportions of vitellogenic oocytes decreased ∼8.8 fold. In males exposed to phenanthrene, spermatogonia proportions were increased ∼2.3 fold in testicular tissues compared to control fish. Thus, gametes were at an earlier stage of maturation in phenanthrene-treated fish compared to controls. However, no differences were detected in the production of 17β-estradiol or testosterone from the gonad in either sex. Catalase activity was also assessed in the liver as a measure of oxidative stress and this biomarker did not change in activity in either sex. In addition to endpoints in the ovary, the female hepatic transcriptome was measured, as this tissue produces lipids for oocyte maturation. Transcriptomic responses to phenanthrene exposure suggested a reduction in vitellogenin mRNA, and lipid metabolism and immune system pathways. Comparisons of hepatic transcriptome responses with Part A (72 h phenanthrene exposure) showed that energy homeostasis pathways were consistently altered following phenanthrene exposure over multiple durations and concentrations. We suggest that altered energy homeostasis may be adversely affecting reproductive efforts, as impaired reproduction has been observed in other studies investigating polycyclic aromatic hydrocarbons., (Copyright © 2018 Elsevier B.V. All rights reserved.)