Development, standardization and refinement of procedures for evaluating effects of endocrine active compounds on development and sexual differentiation of Xenopus laevis

Springer Science and Business Media LLC - Tập 390 - Trang 2031-2048 - 2008
Ilka Lutz1, Werner Kloas1,2, Timothy A. Springer3, Larry R. Holden4, Jeff C. Wolf5, Henry O. Krueger3, Alan J. Hosmer6
1Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Berlin, Germany
2Mathematisch-Naturwissenschaftliche Fakultät I, Institut für Biologie, Berlin, Germany
3Wildlife International, Ltd, Easton, USA
4Sielken and Associates Consulting, Inc., Bryan, USA
5Experimental Pathology Laboratories, Inc., Sterling, USA
6Syngenta Crop Protection, Inc., Greensboro, USA

Tóm tắt

Xenopus laevis has been introduced as a model to study effects of endocrine-active compounds (EAC) on development and sexual differentiation. However, variable and inconsistent data have raised questions about the reliability of the test methods applied. The current study was conducted in two laboratories to develop, refine, and standardize procedures and protocols. Larvae were exposed in flow-through systems to 17β-estradiol (E2), at concentrations from 0.2 to 6.0 μg E2 L−1 in Experiment 1A, and 0.015 to 2.0 μg E2 L−1 in Experiment 1B. In both studies survival (92%, 99%) and percentage of animals that completed metamorphosis (97%, 99%) indicated reproducible biological performance. Furthermore, minor variations in husbandry led to significant differences in snout-to-vent length, weight, and gonad size. In Experiment 1A, almost complete feminization occurred in all E2 treatment groups whereas a concentration response was observed in Experiment 1B resulting in an EC50 of 0.12 μg E2 L−1. The final verified protocol is suitable for determining effects of EAC on development and sexual differentiation in X. laevis.

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