Maria Sepulveda Purdue University marisep@purdue.edu 0000-0003-0473-5760 Maria Sepulveda Purdue University marisep@purdue.edu 0000-0003-0473-5760 Madison Van Slette Arizona State University Undergraduate Research Assistant Jennifer Freeman 0000-0002-3669-9111 Collaborator Ting-Yu Chu Arizona State University Histology Slide Scanner The herbicide atrazine, a suspected endocrine disrupting chemical (EDC), frequently contaminates potable water supplies. Studies suggest alterations in the neuroendocrine system along the hypothalamus-pituitary-gonadal axis; however, most studies address either developmental, pubertal, or adulthood exposures, with few investigations regarding a developmental origins hypothesis. In this study, zebrafish were exposed to 0, 0.3, 3, or 30 parts per billion (ppb) atrazine through embryogenesis and then allowed to mature with no additional chemical exposure. Reproductive function, histopathology, hormone levels, offspring morphology, and the ovarian transcriptome were assessed. Embryonic atrazine exposure resulted in a significant increase in progesterone levels in the 3 and 30ppb groups. A significant decrease in spawning and a significant increase in follicular atresia in the 30ppb group were observed. In offspring, a decrease in the head length to body ratio in the 30ppb group, along with a significant increase in head width to body ratio in the 0.3 and 3ppb groups occurred. Transcriptomic alterations involved genes associated with endocrine system development and function, tissue development, and behavior. This study provides evidence to support atrazine as an EDC causing reproductive dysfunction and molecular alterations in adults exposed only during embryogenesis and morphological alterations in their offspring. danio rerio zebrafish atrazine endocrine disruption ovary follicles 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. 2013 2015 Subphylum Craniata 89593 Superclass Actinopterygii ray-finned fishes 7898 Class Actinopteri 186623 Subclass Neopterygii 41665 Infraclass Teleostei teleost fishes 32443 Superorder Cypriniphysae 186627 Order Cypriniformes carps and others 7952 Suborder Cyprinoidei 30727 Family Danionidae 2743709 Subfamily Danioninae 2743711 Genus Danio 7954 Species Danio rerio zebrafish 7955 Maria Sepulveda Purdue University marisep@purdue.edu 0000-0003-0473-5760 Adult female zebrafish were euthanized in buffered MS-222 (Ethyl 3-aminobenzoate methanesulfonate, Sigma, St. Louis, MO) (4 mg/mL). A sample of ovarian tissue was collected from both ovaries and fixed in Davidson’s Fixative overnight at room temperature and then transferred to histology grade 70% EtOH (Sigma). Hematoxylin and eosin (H & E) sections were prepared following standard procedures and sections examined under a light microscope (10–40X) for staging and evaluation of any abnormalities. All follicles were counted and classified as perinuclear, cortical alveoli, early vitellogenic, late vitellogenic, post-ovulatory, or atretic. Frequencies of occurrence of each stage were then calculated from each section and averaged. MS_DR_1B_003_7 MS_DR_1B_003_7.ome.tif ome.tif Fish ID: 1B-7; Life Stage: 3-reproductively mature; Mature oocytes: 43; Immature oocytes: 52; Perinucleolar oocytes: 58; Atretic oocytes: 2; Total oocytes: 155; Publication: Wirbisky SE, Weber GJ, Sepúlveda MS, Lin TL, Jannasch AS, Freeman JL. An embryonic atrazine exposure results in reproductive dysfunction in adult zebrafish and morphological alterations in their offspring. Sci Rep. 2016 Feb 19; 6:21337. doi: 10.1038/srep21337. PMID: 26891955; PMCID: PMC4759560. histology MS_DR_1B_003_7 female adult 1 ovary unspecified 003_7 Atrazine, 0.3 ug/L, aqueous, exposed as embryos 1-72 hours post fertilization (hpf) 5 microns Davidson’s Fixative overnight at room temperature and then transferred to histology grade 70% EtOH Olympus CX33