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2L - Immunotoxicity - genotoxicity - ED (WEP/130) Application of Hybridisation Protection Assays for studying endocrine disruption in fish. Riley, Gavin1, Tyler, Charles1, Thomas-Jones, Emma2, 1 University of Exeter, Exeter, UK, United Kingdom2 Molecular Light Technology, Cardiff, Wales, United Kingdom ABSTRACT- It is now widely acknowledged that there are chemicals present in the environment that can interfere with hormone function and cause detrimental effects on the physiology of organisms, from invertebrates to humans. Most reported examples of endocrine disruption are alterations in reproductive function. Altered sexual reproduction as a consequence of endocrine disruption is now widely reported in fish. Furthermore, fish have been instrumental in developing our understanding of the mechanisms of sexual disruption as a consequence of endocrine disruption. Biochemical, cell and tissue-level responses have predominated in studies on endocrine disruption and molecular analysis has been lacking. A battery of techniques for molecular analyses is now available ranging from Q-PCR for quantifying single gene responses to arrays (for multiple gene responses). This presentation details the development of a Hybridisation Protection Assay - that utilises a chemiluminescent-labelled oligonucleotide probe - for quantifying responses of specific mRNA targets to the effects of endocrine disrupting chemicals. The technique quantifies specific transcript abundance directly from total RNA isolates without the requirement for any initial amplification step. Data on the use of the HPA system in the study of vitelline envelope protein responses to 17 Key words: Hybridisation Protection Assay, Endocrine Disruption, Fathead Minnow, Rainbow Trout |
-oestradiol and 17
ethinyl oestradiol in the rainbow trout (Oncorhynchus mykiss) will be presented together with validation work for probes designed to target aromatase P450 (A) and (B) in the fathead minnow (Pimephales promelas).