title: Proteomic analysis of zebrafish brain tissue following exposure to the pesticide prochloraz.
abstract: The hypothalamus-pituitary-gonadal (HPG) axis plays a central role in the maintenance of homeostasis and disruptions in its function can have important implications for reproduction and other critical biological processes. A number of compounds found in aquatic environments are known to affect the HPG axis. In the present study, we used two-dimensional electrophoresis to investigate the proteome of female and male zebrafish brain after 96 h exposure to the fungicide prochloraz. Prochloraz has known effects on a number of key HPG molecules, including antagonism of Cyp17 and Cyp19 (aromatase). Twenty-eight proteins were shown to be differentially expressed in the brains of females and 22 in males. Proteins were identified using LC-MS/MS and identities were examined relative to brain function in the context of changing steroid hormone levels. There was little overlap between sexes in proteins exhibiting differential expression. Proteins with known roles in metabolism, learning, neuroprotection, and calcium regulation were determined to be differentially regulated. Relationships between identified proteins were also examined using Ingenuity Pathway Analysis, and females were shown to exhibit enrichment of several metabolic pathways. We used differentially expressed proteins to establish a putative classifier consisting of three proteins that was able to discriminate prochloraz-exposed from control females. Putatively impacted brain functions and specific protein changes that were observed have the potential to be generalized to other that similarly impact steroid hormone levels.
authors.affiliation: Environmental Protection Agency, Office Research and Development, National Exposure Research Laboratory, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA. biales.adam@epa.gov
authors.forename: Adam D
authors.initials: AD
authors.lastname: Biales
authors.forename: David C
authors.initials: DC
authors.lastname: Bencic
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Villeneuve
authors.forename: Gerald T
authors.initials: GT
authors.lastname: Ankley
authors.forename: David L
authors.initials: DL
authors.lastname: Lattier
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004703:Endocrine System / Q000187:drug effects; D005260:Female; D029941:Fish Proteins / Q000378:metabolism; D005659:Fungicides, Industrial / Q000633:toxicity*; D006728:Hormones / Q000378:metabolism; D007030:Hypothalamo-Hypophyseal System / Q000187:drug effects; D007093:Imidazoles / Q000633:toxicity*; D008297:Male; D025341:Principal Component Analysis; D020543:Proteome / Q000187:drug effects*; D040901:Proteomics; D011897:Random Allocation; D012737:Sex Factors; D053719:Tandem Mass Spectrometry; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D029941:Fish Proteins; D005659:Fungicides, Industrial; D006728:Hormones; D007093:Imidazoles; D020543:Proteome; D014874:Water Pollutants, Chemical; C045362:prochloraz