In situ hybridization to detect spatial gene expression in medaka

A. R. TOMPSETT, J. W. PARK, X. ZHANG, P. D. JONES, J. L. NEWSTED, D. W. T. AU, E. X. H. CHEN, R. YU, Shiu Sun Rudolf WU, R. Y. C. KONG, J. P. GIESY, M. HECKER

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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 languageEnglish
Pages (from-to)1257-1264
JournalEcotoxicology and Environmental Safety
Volume72
Issue number4
DOIs
Publication statusPublished - May 2009

Fingerprint

Oryzias
Fadrozole
Gene expression
In Situ Hybridization
Tissue
Gene Expression
Animals
Oviparity
Aromatase Inhibitors
Aromatase
Polymerase chain reaction
Radio
Fish
Oocytes
Testis
Real-Time Polymerase Chain Reaction
Ovary
Fishes
Animal Models
Genes

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.2008.10.013

Keywords

  • Histology
  • Fadrozole
  • Aromatase
  • CYP19
  • Hormones
  • Autoradiography
  • Oryzias latipes
  • RT-PCR
  • Gene expression
  • Endocrine disruption