An approach to detecting estrogenic endocrine disruption via choriogenin expression in an estuarine model fish...

A large body of work has established a link between endocrine disrupting compounds (EDCs) and a number of abnormalities in fishes. However, most EDC studies use several standard laboratory denizens to assess impacts, so assumptions about sensitivity are primarily based on these few species. Additionally, existing methods rely on obtaining sufficient plasma to measure EDC biomarkers. Our objectives were (a) to establish a new model species for estuarine fishes, (b) to evaluate endocrine impacts with a highly sensitive and specific biomarker, and (c) to develop a method for the analysis of this biomarker in small fish that do not possess sufficient blood plasma for protein measurement. As such, we created a polyclonal antibody (Ab) to the estrogen-responsive proteins chorion (Ch) and choriogenin (Chg) in Menidia beryllina, found throughout coastal North America and already utilized in EPA Whole Effluent Testing. We then validated the Ab by using it to measure the response to aqueous ethinylestradiol (EE2) through the development an ELISA using Menidia whole body homogenate (WBH). Sensitivity of the Ab to Menidia WBH is greater than that of the commercially available option. ELISA sensitivity, with a detection limit of 5 ng/ml and a working range of 22.6-1370.9 ng/ml, is comparable to ELISAs developed to measure plasma Chg. To our knowledge this is the first ELISA method developed for the detection of Chg using WBH. Including additional model species and methods allowing the evaluation of alternative sample matrices will contribute to an enhanced understanding of inter-species differences in EDC response.