title: Exposure to atrazine affects the expression of key genes in metabolic pathways integral to energy homeostasis in Xenopus laevis tadpoles.
abstract: In our laboratory, Xenopus laevis tadpoles exposed throughout development to 200 or 400 μg/L atrazine, concentrations reported to periodically occur in puddles, vernal ponds and runoff soon after application, were smaller and had smaller fat bodies (the tadpole's lipid storage organ) than controls. It was hypothesized that these changes were due to atrazine-related perturbations of energy homeostasis. To investigate this hypothesis, selected metabolic responses to exposure at the transcriptional and biochemical levels in atrazine-exposed tadpoles were measured. DNA microarray technology was used to determine which metabolic pathways were affected after developmental exposure to 400 μg/L atrazine. From these data, genes representative of the affected pathways were selected for assay using quantitative real time polymerase chain reaction (qRT-PCR) to measure changes in expression during a 2-week exposure to 400 μg/L. Finally, ATP levels were measured from tadpoles both early in and at termination of exposure to 200 and 400 μg/L. Microarray analysis revealed significant differential gene expression in metabolic pathways involved with energy homeostasis. Pathways with increased transcription were associated with the conversion of lipids and proteins into energy. Pathways with decreased transcription were associated with carbohydrate metabolism, fat storage, and protein synthesis. Using qRT-PCR, changes in gene expression indicative of an early stress response to atrazine were noted. Exposed tadpoles had significant decreases in acyl-CoA dehydrogenase (AD) and glucocorticoid receptor protein (GR) mRNA after 24 h of exposure, and near-significant (p=0.07) increases in peroxisome proliferator-activated receptor β (PPAR-β) mRNA by 72 h. Decreases in AD suggested decreases in fatty acid β-oxidation while decreases in GR may have been a receptor desensitization response to a glucocorticoid surge. Involvement of PPAR-β, an energy homeostasis regulatory molecule, also suggested changes in energy status. Despite, or possibly because of, these early gene changes, there were no differences in either absolute ATP levels or ADP:ATP ratios early in the exposure. However, livers from animals exposed to 200 μg/L atrazine had near-significant (p=0.06) increases in ADP:ATP ratios at the end of exposure suggesting tadpoles may have had difficulty maintaining energy homeostasis. Perturbations in the expression of genes regulating energy metabolism by 24 h into exposure to 400 μg/L atrazine was noteworthy, especially since these tadpoles were significantly smaller than controls by 72 h of exposure.
authors.affiliation: Great Lakes Environmental and Molecular Sciences Center, Department of Biological Sciences, Western Michigan University, Kalamazoo, MI 49008, USA. renee.zaya@wmich.edu
authors.forename: Renee M
authors.initials: RM
authors.lastname: Zaya
authors.forename: Zakariya
authors.initials: Z
authors.lastname: Amini
authors.forename: Ashley S
authors.initials: AS
authors.lastname: Whitaker
authors.forename: Charles F
authors.initials: CF
authors.lastname: Ide
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001280:Atrazine / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects*; D006540:Herbicides / Q000633:toxicity*; D006706:Homeostasis / Q000187:drug effects; D007814:Larva / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D053858:Metabolic Networks and Pathways / Q000187:drug effects*; D020411:Oligonucleotide Array Sequence Analysis; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D014982:Xenopus laevis / Q000378:metabolism / Q000502:physiology*
chemical_list: D006540:Herbicides; D014874:Water Pollutants, Chemical; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D001280:Atrazine