Abstract
A whole-animal tissue section in situ hybridization (ISH) system with radio-labeled probes was developed to detect differential gene expression among tissues of the small, oviparous teleost fish, Japanese medaka (Oryzias latipes). Because of its tissue- and gender-specific expression, gonadal aromatase (CYP19a) was selected as a model gene to demonstrate the potential of the system. The ISH system was validated with a 7 d exposure to the model aromatase inhibitor, fadrozole. Fadrozole did not affect the magnitude of gene expression in testes, but significantly up-regulated CYP19a gene expression in ovaries. These results were confirmed with quantitative real-time-polymerase chain reaction (RT-PCR). Histological evaluation revealed that females exposed to 100 μg/L fadrozole lacked mature oocytes. Male gonadal morphology was normal in all treatments. The ISH method developed in this study allowed tissue-specific resolution of gene expression in a whole animal model, as well as the ability to analyze cellular morphological detail in the same organism. Copyright © 2008 Elsevier Inc. All rights reserved.
Original language | English |
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Pages (from-to) | 1257-1264 |
Journal | Ecotoxicology and Environmental Safety |
Volume | 72 |
Issue number | 4 |
DOIs | |
Publication status | Published - May 2009 |
Citation
Tompsett, A. R., Park, J. W., Zhang, X., Jones, P. D., Newsted, J. L., Au, D. W. T., . . . Hecker, M. (2009). In situ hybridization to detect spatial gene expression in medaka. Ecotoxicology and Environmental Safety, 72(4), 1257-1264. doi: 10.1016/j.ecoenv.2008.10.013Keywords
- Histology
- Fadrozole
- Aromatase
- CYP19
- Hormones
- Autoradiography
- Oryzias latipes
- RT-PCR
- Gene expression
- Endocrine disruption