Quantitative measurement of fathead minnow vitellogenin mRNA using hybridization protection assays.
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 17beta-estradiol demonstrating the utility of this technique for the identification and monitoring of toxic responses to xenobiotics.
Molecular Light Technology Research Limited, 5 Chiltern Close, Cardiff Industrial Park, Cardiff CF14 5DL, Wales, United Kingdom.
Emma
E
Thomas-Jones
Neal
N
Walkley
Ceri
C
Morris
Peter
P
Kille
Jennifer
J
Cryer
Ian
I
Weeks
J Stuart
JS
Woodhead
D000818:Animals; D003530:Cyprinidae / Q000235:genetics / Q000378:metabolism*; D004781:Environmental Exposure / Q000032:analysis; D004784:Environmental Monitoring / Q000379:methods*; D004958:Estradiol / Q000494:pharmacology / Q000633:toxicity; D007274:Injections, Intraperitoneal; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000032:analysis / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D014158:Transcription, Genetic / Q000187:drug effects*; D014819:Vitellogenins / Q000096:biosynthesis* / Q000235:genetics
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D014819:Vitellogenins; D004958:Estradiol