Quantitative measurement of fathead minnow vitellogenin mRNA using hybridization protection assays

• Published in: Environmental toxicology and chemistry 2003-05, Vol.22 (5), p.992-995
• Main Authors: Thomas-Jones, Emma, Walkley, Neal, Morris, Ceri, Kille, Peter, Cryer, Jennifer, Weeks, Ian, Woodhead, J. Stuart
• Format: Article
• Language: English
• Subjects: Agnatha. Pisces; Animal, plant and microbial ecology; Animals; Applied ecology; Biological and medical sciences; Cyprinidae - genetics; Cyprinidae - metabolism; Ecotoxicology, biological effects of pollution; Effects of pollution and side effects of pesticides on vertebrates; Endocrine disruption; Environmental Exposure - analysis; Environmental Monitoring - methods; Estradiol - pharmacology; Estradiol - toxicity; estrogenic activity; Fathead minnow; Freshwater; Fundamental and applied biological sciences. Psychology; Gene expression; Hybridization protection assay; Injections, Intraperitoneal; Luminescent Measurements; Nucleic Acid Hybridization - methods; Oligonucleotide Probes - analysis; Oligonucleotide Probes - chemistry; Pimephales promelas; Reproducibility of Results; RNA, Messenger - analysis; RNA, Messenger - genetics; Techniques; Transcription, Genetic - drug effects; Vitellogenin; Vitellogenins - biosynthesis; Vitellogenins - genetics
• ISSN: 0730-7268; 1552-8618
• DOI: 10.1002/etc.5620220506

We have developed a novel test system for the quantitative assessment of gene transcription. The procedure involves the use of chemiluminescent‐labeled oligonucleotide probes in a hybridization protection assay (HPA) format. We have used this technology to measure changes in vitellogenin mRNA to demonstrate the impact of estrogen exposure in the juvenile fathead minnow (Pimephales promelas). Marked changes in mRNA expression were observed in response to intraperitoneal injection of 17β‐estradiol demonstrating the utility of this technique for the identification and monitoring of toxic responses to xenobiotics.