In situ

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• 作者：A.R. Tompsett ; J.W. Park ; X. Zhang ; P.D. Jones ; J.L. Newsted ; D.W.T. Au ; E.X.H. Chen ; R. Yu ; R.S.S. Wu ; R.Y.C. Kong ; J.P. Giesy ; M. Hecker
• 关键词：Histology ; Fadrozole ; Aromatase ; CYP19 ; Hormones ; Autoradiography ; Oryzias latipes ; RT-PCR ; Gene expression ; Endocrine disruption
• 刊名：Ecotoxicology and Environmental Safety
• 出版年：2009
• 出版时间：May 2009
• 年：2009
• 卷：72
• 期：4
• 页码：1257-1264
• 全文大小：1878 K

文摘

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.