title: Atrazine exposure elicits copy number alterations in the zebrafish genome.
abstract: Atrazine is an agricultural herbicide used throughout the Midwestern United States that frequently contaminates potable water supplies resulting in human exposure. Using the zebrafish model system, an embryonic atrazine exposure was previously reported to decrease spawning rates with an increase in progesterone and ovarian follicular atresia in adult females. In addition, alterations in genes associated with distinct molecular pathways of the endocrine system were observed in brain and gonad tissue of the adult females and males. Current hypotheses for mechanistic changes in the developmental origins of health and disease include genetic (e.g., copy number alterations) or epigenetic (e.g., DNA methylation) mechanisms. As such, in the current study we investigated whether an atrazine exposure would generate copy number alterations (CNAs) in the zebrafish genome. A zebrafish fibroblast cell line was used to limit detection to CNAs caused by the chemical exposure. First, cells were exposed to a range of atrazine concentrations and a crystal violet assay was completed, showing confluency decreased by ~60% at 46.3μM. Cells were then exposed to 0, 0.463, 4.63, or 46.3μM atrazine and array comparative genomic hybridization completed. Results showed 34, 21, and 44 CNAs in the 0.463, 4.63, and 46.3μM treatments, respectively. Furthermore, CNAs were associated with previously reported gene expression alterations in adult male and female zebrafish. This study demonstrates that atrazine exposure can generate CNAs that are linked to gene expression alterations observed in adult zebrafish exposed to atrazine during embryogenesis providing a mechanism of the developmental origins of atrazine endocrine disruption.
authors.affiliation: School of Health Sciences, Purdue University, West Lafayette, IN, 47909, United States. Electronic address: swirbisk@purdue.edu.
authors.forename: Sara E
authors.initials: SE
authors.lastname: Wirbisky
authors.affiliation: School of Health Sciences, Purdue University, West Lafayette, IN, 47909, United States. Electronic address: jfreema@purdue.edu.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Freeman
mesh_terms: D000818:Animals; D001280:Atrazine / Q000633:toxicity*; D001921:Brain / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D002460:Cell Line; D055028:Comparative Genomic Hybridization; D047108:Embryonic Development / Q000187:drug effects; D052244:Endocrine Disruptors / Q000633:toxicity; D005260:Female; D018628:Gene Dosage / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D016678:Genome / Q000187:drug effects*; D006540:Herbicides / Q000633:toxicity*; D008297:Male; D009153:Mutagens / Q000633:toxicity*; D009994:Osmolar Concentration; D010053:Ovary / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013737:Testis / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000633:toxicity; D015027:Zebrafish / Q000196:embryology / Q000502:physiology*
chemical_list: D052244:Endocrine Disruptors; D006540:Herbicides; D009153:Mutagens; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical; D001280:Atrazine
keywords: Array comparative genomic hybridization; Atrazine; Copy number alterations; Transcriptomics; Zebrafish