Custom microarray construction and analysis for determining potential biomarkers of subchronic androgen exposure in the Eastern Mosquitofish (Gambusia holbrooki)

BMC Genomics

Research Article

Erica K Brockmeier¹  Nancy D Denslow²  David Moraga Amador³  Fahong Yu³  Timothy A Bargar⁴ 

[1] Department of Physiological Sciences, Center for Environmental and Human Toxicology, University of Florida, P.O. Box 110885, 2187 Mowry Road, 32611, Gainesville, FL, USA;Department of Physiological Sciences, Center for Environmental and Human Toxicology, University of Florida, P.O. Box 110885, 2187 Mowry Road, 32611, Gainesville, FL, USA;Genetics Institute, University of Florida, P.O. Box 103610, 2033 Mowry Road, 32610, Gainesville, FL, USA;Interdisciplinary Center for Biotechnology Research, University of Florida, P.O. Box 103622, 32611, Gainesville, FL, USA;Southeast Ecological Science Center, U.S. Geological Survey, 7920 NW 71st Street, 32653, Gainesville, FL, USA;

关键词: Gambusia holbrooki;    17β-trenbolone;    Androgen biomarker;    Aquatic toxicology;    Ecotoxicogenomics;    Microarray;    Gonopodium;   

DOI  :  10.1186/1471-2164-14-660

 received in 2013-06-26, accepted in 2013-09-25,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundThe eastern mosquitofish (Gambusia holbrooki) has the potential to become a bioindicator organism of endocrine disrupting chemicals (EDCs) due to its androgen-driven secondary sexual characteristics. However, the lack of molecular information on G. holbrooki hinders its use as a bioindicator coupled with biomarker data. While traditional gene-by-gene approaches provide insight for biomarker development, a holistic analysis would provide more rapid and expansive determination of potential biomarkers. The objective of this study was to develop and utilize a mosquitofish microarray to determine potential biomarkers of subchronic androgen exposure. To achieve this objective, two specific aims were developed: 1) Sequence a G. holbrooki cDNA library, and 2) Use microarray analysis to determine genes that are differentially regulated by subchronic androgen exposure in hepatic tissues of 17β-trenbolone (TB) exposed adult female G. holbrooki.ResultsA normalized library of multiple organs of male and female G. holbrooki was prepared and sequenced by the Illumina GA IIx and Roche 454 XLR70. Over 30,000 genes with e-value ≤ 10-4 were annotated and 14,758 of these genes were selected for inclusion on the microarray. Hepatic microarray analysis of adult female G. holbrooki exposed to the vehicle control or 1 μg/L of TB (a potent anabolic androgen) revealed 229 genes upregulated and 279 downregulated by TB (one-way ANOVA, p < 0.05, FDR α = 0.05, fold change > 1.5 and < −1.5). Fifteen gene ontology biological processes were enriched by TB exposure (Fisher’s Exact Test, p < 0.05). The expression levels of 17β-hydroxysteroid dehydrogenase 3 and zona pellucida glycoprotein 2 were validated by quantitative polymerase chain reaction (qPCR) (Student’s t-test, p < 0.05).ConclusionsCoupling microarray data with phenotypic changes driven by androgen exposure in mosquitofish is key for developing this organism into a bioindicator for EDCs. Future studies using this array will enhance knowledge of the biology and toxicological response of this species. This work provides a foundation of molecular knowledge and tools that can be used to delve further into understanding the biology of G. holbrooki and how this organism can be used as a bioindicator organism for endocrine disrupting pollutants in the environment.

【 授权许可】

Unknown   
© Brockmeier et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.