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Michael E. Baker,¹ Barbara Ruggeri,^2,3 L. James Sprague,² Colleen Eckhardt-Ludka,² Jennifer Lapira,² Ivan Wick,² Laura Soverchia,^2,3 Massimo Ubaldi,³ Alberta Maria Polzonetti-Magni,³ Doris Vidal-Dorsch,⁴ Steven Bay,⁴ Joseph R. Gully,⁵ Jesus A. Reyes,⁶ Kevin M. Kelley,⁶ Daniel Schlenk,⁷ Ellen C. Breen,⁸ Roman Šášik,^2,9 and Gary Hardiman^1,2

¹Department of Medicine and ²BioMedical Genomics Facility, School of Medicine, University of California, San Diego, La Jolla, California, USA; ³Department of Experimental Medicine and Public Health, University of Camerino, Camerino, Italy; ⁴Southern California Coastal Water Research Project, Costa Mesa, California, USA; ⁵Los Angeles County Sanitation Districts, Whittier, California, USA; ⁶Environmental Endocrinology Laboratory, California State University, Long Beach, California, USA; ⁷Department of Environmental Sciences, University of California, Riverside, California, USA; ⁸Division of Physiology, and ⁹Moore's Cancer Center, University of California, San Diego, La Jolla, California, USA

Abstract
Background: Endocrine disruptors include plasticizers, pesticides, detergents, and pharmaceuticals. Turbot and other flatfish are used to characterize the presence of chemicals in the marine environment. Unfortunately, there are relatively few genes of turbot and other flatfish in GenBank, which limits the use of molecular tools such as microarrays and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) to study disruption of endocrine responses in sentinel fish captured by regulatory agencies.

Objectives: We fabricated a multigene cross-species microarray as a diagnostic tool to screen the effects of environmental chemicals in fish, for which there is minimal genomic information. The array included genes that are involved in the actions of adrenal and sex steroids, thyroid hormone, and xenobiotic responses. This microarray will provide a sensitive tool for screening for the presence of chemicals with adverse effects on endocrine responses in coastal fish species.

Methods: We used a custom multispecies microarray to study gene expression in wild hornyhead turbot (Pleuronichthys verticalis) collected from polluted and clean coastal waters and in laboratory male zebrafish (Danio rerio) after exposure to estradiol and 4-nonylphenol. We measured gene-specific expression in turbot liver by qRT-PCR and correlated it to microarray data.

Results: Microarray and qRT-PCR analyses of livers from turbot collected from polluted areas revealed altered gene expression profiles compared with those from nonaffected areas.

Conclusions: The agreement between the array data and qRT-PCR analyses validates this multispecies microarray. The microarray measurement of gene expression in zebrafish, which are phylogenetically distant from turbot, indicates that this multispecies microarray will be useful for measuring endocrine responses in other fish.

Key words: Danio rerio, endocrine disruptors, flatfish, hornyhead turbot, microarray, multispecies array, nonylphenol, Pleuronichthys verticalis, xenobiotics, xenoestrogens, zebrafish. Environ Health Perspect 117:223–230 (2009) . doi:10.1289/ehp.11627 available via http://dx.doi.org/ [Online 28 August 2008]

Address correspondence to G. Hardiman, BIOGEM, School of Medicine, Department of Medicine, University of California San Diego, 9500 Gilman Dr., MC 0724, La Jolla, CA 92093 USA. Telephone: (858) 822-3792. Fax: (858) 822-6430. E-mail: ghardiman@ucsd.edu

Supplemental Material is available online at http://www.ehponline.org/members/2008/11627/suppl.pdf

The multispecies microarray was jointly developed in the laboratories of M.E.B. and G.H. We thank members of both laboratories for useful discussions. We also thank the Los Angeles County Sanitation Districts and Orange County Sanitation District for their assistance.

Partial funding for this project was provided by the Los Angeles County Sanitation Districts. BIOGEM was supported by National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases Award 1 P30 DK063491-03, a contract from the Orange County Sanitation District and the Los Angeles County Sanitation Districts administered via Southern California Coastal Water Research Project and University of Southern California Sea Grant Program (National Oceanic and Atmospheric Administration grant NA06OAR4170012, Catalog of Federal Domestic Assistance no. 11.417, project CE-17) .

The authors declare they have no competing financial interests.

Received 25 April 2008 ; accepted 27 August 2008.

Correction

Figure 7 was incorrect in the original article published online. It has been corrected here.