Chad A Blanksma US EPA Great Lakes Toxicology and Ecology Division Histotechnologist

6201 Congdon Blvd. Duluth MN 55804 USA

blanksma.chad@epa.gov 0009-0001-8811-8083 An image of an individual vitellogenic oocyte from an adult fathead minnow. In the germinal vesicle, nucleoli and lampbrush chromosomes can be observed. Areas of cortical alveolar oocytes along with perinucleolar oocytes, a post-ovulatory follicle, and pigmented macrophage aggregates can also be found in this micrograph. Vitellogenic oocyte Germinal vesicle Nucleoli Lampbrush chromosomes Perinucleolar oocyte Cortical alveolar oocyte Post-ovulatory follicle Pigmented macrophage aggregate Adult Fathead minnow This information is released under the Creative Commons license - Attribution - CC BY (https://creativecommons.org/licenses/by/4.0/). The consumer of these data ("Data User" herein) is required to cite it appropriately in any publication that results from its use. The Data User should realize that these data may be actively used by others for ongoing research and that coordination may be necessary to prevent duplicate publication. The Data User is urged to contact the authors of these data if any questions about methodology or results occur. Where appropriate, the Data User is encouraged to consider collaboration or co-authorship with the authors. The Data User should realize that misinterpretation of data may occur if used out of context of the original study. While substantial efforts are made to ensure the accuracy of data and associated documentation, complete accuracy of data sets cannot be guaranteed. All data are made available "as is." The Data User should be aware, however, that data are updated periodically and it is the responsibility of the Data User to check for new versions of the data. The data authors and the repository where these data were obtained shall not be liable for damages resulting from any use or misinterpretation of the data. Thank you. 2012 Kingdom Animalia animals 202423 Subkingdom Bilateria triploblasts 914154 Infrakingdom Deuterostomia 914156 Phylum Chordata chordates 158852 Subphylum Vertebrata vertebrates 331030 Infraphylum Gnathostomata 914179 Superclass Actinopterygii ray-finned fishes 161061 Class Teleostei 161105 Superorder Ostariophysi 162845 Order Cypriniformes minnows 162846 Superfamily Cyprinoidea 638796 Family Leuciscidae 1198935 Subfamily Pogonichthyinae 1198937 Genus Pimephales bluntnose minnows 163515 Species Pimephales promelas Fathead Minnow 163517 Chad A Blanksma US EPA Great Lakes Toxicology and Ecology Division Histotechnologist

Duluth MN 55804 USA

blanksma.chad@epa.gov 0009-0001-8811-8083 Image collection device Diagnostic Instruments, Inc. SPOT Insight 4 Mp CCD Scientific Color Digital Camera Image collection software Diagnostic Instruments, Inc. SPOT Advanced Software Image analysis software Media Cybernetics Image-Pro Premier Graphics Editing Software Corel Corporation Corel Photo-Paint Sarah Y Skolness

Department of Biology, University of Nebraska at Omaha 6001 Dodge St. Omaha NE 68182 USA

Corresponding author Conazoles are designed to inhibit cytochrome P450 (CYP) 14α-demethylase, an enzyme key to fungal cell wall formation. In vertebrates, conazoles may inhibit other CYPs, potentially disrupting processes like sex steroid synthesis. Propiconazole is a current-use pesticide that is among the first chemicals being tested in the U.S. Environmental Protection Agency endocrine disruptor screening program. Fathead minnows (Pimephales promelas) were exposed to 0, 5, 50, 500, or 1000 µg propiconazole/l in a 21-day study that evaluated apical reproductive endpoints (fecundity, fertility, hatch); measures of endocrine function and steroid synthesis, such as cholesterol, vitellogenin (VTG), and sex steroid (testosterone [T], 17β-estradiol [E2]) concentrations in the plasma; and changes in gonadal expression of steroidogenic genes. Plasma E2 and VTG concentrations in females were reduced by exposure to propiconazole, and egg production was decreased in the 500 and 1000 µg/l treatment groups. These in vivo effects coincided with inhibition of E2 synthesis by ovary explants exposed to propiconazole in vitro. We also observed a compensatory response in females exposed to propiconazole, manifested as increased gonad weight and upregulation of genes coding for key steriodogenic proteins, including CYP19 (aromatase), CYP17 (hydroxylase/lyase), CYP11A (cholesterol side-chain-cleavage), and steroidogenic acute regulatory protein. Other than an increase in relative testis weight, effects on endocrine function in males were less pronounced than in females. This study provides important data relative to the potential endocrine activity of propiconazole in fish and, more generally, to the further delineation of pathways for the reproductive effects of steroid synthesis inhibitors in fish. Skolness SY, Blanksma CA, Cavallin JE, Churchill JJ, Durhan EJ, Jensen KM, Johnson RD, Kahl MD, Makynen EA, Villeneuve DL, Ankley GT. Propiconazole inhibits steroidogenesis and reproduction in the fathead minnow (Pimephales promelas). Toxicol Sci. 2013 Apr;132(2):284-97. doi: 10.1093/toxsci/kft010. Epub 2013 Jan 20. PMID: 23339182. C-2_mid-vitellogenic_oocyte_20x C-2_mid-vitellogenic_oocyte_20x.tif tif histology USEPA_GLTED_PP_Propi21d_CA2 female adult 1 ovary unspecified 1 4 microns Zeiss Axiovert 35