W9 AM Biomonitoring
Wednesday, 16 November 2005: 8:00 AM - 11:40 AM in 339-340

(VAN-1117-803845) A simple, universal assay for vitellogenin in fish mucus and blood.

Van Veld, P¹, Fisher, D², Johnston, L¹, Rice, C³, Rutan, B¹, Sullivan, C¹, Yonkos, L², ¹ College of William and Mary, Virginia Institute of Marine Science, Gloucester Point, VA, USA² University of Maryland, Wye Research and Education Center, Queenstown, MD, USA³ Clemson University, Department of Biological Sciences, Pendleton, SC, USA

ABSTRACT- Vitellogenins are high molecular weight phosphoproteins that serve as precursors to egg yolk proteins in all oviparous and ovoviviparous vertebrates. Though normally produced in females, expression of vitellogenin in male fish has become a widely used biomarker of exposure to environmental estrogens. Vitellogenin is usually measured in blood by antibody-based methods that often require development of species-specific antibodies. In the present paper we describe a universal assay that is based on high molecular weight and extensive phosphoserine content of mucosal and plasma vitellogenins. Blood and mucosal proteins were separated by electrophoresis in polyacrylamide gels, stained with a commercially available fluropore dye and visualized on a uv transilluminator. The method allowed sensitive detection of vitellogenin in females and estradiol-treated males in all species tested. Quantitative analysis indicated that the phosphoprotein assay was at least as sensitive as antibody-based methods but offers the advantage of performing analyses on multiple species simultaneously.

Key words: vitellogenin, universal assay, fish, mucus blood