PM21 New Advances in Estimating Effects of Toxicants on Populations
Exhibit Hall
8:00 AM - Monday
(PM304) Use of Vitellogenin mRNA QRT-PCR for Monitoring Endocrine Disruption in Feral and Cultured Fish.
Pina, Benjamin1, Quiros, L1, Casado, M1, Garcia-Reyero, N1, Raldua, D2, Cerda, J2, Barcelo, D3, 1 IBMB-CSIC, Barcelona, Spain2 Institut for Aquaculture - IRTA, St Carles de la Rapita, Barcelona, Spain3 IIQAB-CSIC, Barcelona, Spain
ABSTRACT- The presence of the female-specific yolk protein precursor vitellogenin in blood and liver from male fish is widely used as an indicator for endocrine disruption. We studied the induction of vitellogenin mRNA in liver from several species of fish, both maintained in fish tanks or captured in the wild using Quantitative Real-Time PCR (QRT-PCR). Our protocol requires minute amounts of liver samples (down to 50 mg), and can be applied to field samples, provided he appropriated RNA-stabilizing agent is used. QRT-PCR proved able to precise quantify vitellogenine mRNA levels in mummichog (Fundulus heteroclitus) and common carp (Cyprinus carpio), with a much higher sensitivity than the standard technique of immunodetection of the vitellogenin protein. In fish natural populations, the system is able no only to distinguish undisturbed males from contaminated ones and from females, but also between winter and summer populations. We are currently exploring expression of other environmentally significant genes such as Metallothionein or Cytochrome P450 with the same technique The use of mRNA quantitation techniques for the analysis of feral and cultured fishes from different species opens the possibility of a much precise detection, and further control, of the noxious effects of contaminants on the local fauna exposed to them.
Key words: endocrine disruption, quantitative real Time pcr, fish ecotoxicology, vitellogenin mrna