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Toxicological Sciences 57, 264-274 (2000)
Copyright © 2000 by the Society of Toxicology

Reproductive and Developmental Toxicology

Stage- and Species-Specific Developmental Toxicity of All-Trans Retinoic Acid in Four Native North American Ranids and Xenopus laevis

Sigmund J. Degitz1, Patricia A. Kosian, Elizabeth A. Makynen, Kathleen M. Jensen and Gerald T. Ankley

U.S. Environmental Protection Agency, Mid-Continent Ecology Division, 6201 Congdon Boulevard, Duluth, Minnesota 55804

Within the last decade, there have been increasing reports of malformed amphibians across North America. Recently, it has been suggested that hind-limb malformations are a consequence of xenobiotic disruption of developmental pathways regulated by retinoids. To assess the validity of this hypothesis, the developmental toxicity of all-trans retinoic acid (RA) was examined in Xenopus laevis and four North American anurans, at several life stages. To determine the effects of RA on embryogenesis, mid-blastula stage embryos were exposed to 0, 6.25, 12.5, 25, or 50 ng RA/ml for 24 h. To evaluate the effects of RA on hind-limb development, early- and mid-limb bud stage tadpoles were exposed to RA concentrations of 0, 250, 500, 750, 1000, or 1250 ng RA/ml for 24 h. Mid-blastula RA exposure resulted in a concentration-dependent increase in dysmorphogenesis and mortality in the three species examined (R. clamitans, R. septentrionalis and X. laevis). RA exposure at stage 51 in X. laevis and stage 28 in R. sylvatica resulted in concentration-dependent increases in reductions and deletions of the hind limb. However, RA was ineffective at inducing hind-limb abnormalities in stages 26 and 28 of R. pipiens, stage 28 in R. clamitans, or stage 48 in X. laevis tadpoles. These results indicate that mid-blastula stage embryos are more sensitive to RA-induced dysmorphogenesis and mortality than limb-bud stage tadpoles. The significance of these findings is discussed in the context of the possible occurrence of retinoid mimics in the environment.

Key Words: all-trans retinoic acid (RA); hindlimb malformation; Rana clamitans; Rana pipiens; Rana septentrionalis; Rana sylvatica; Xenopus laevis.


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