Evaluation of the effect of acetochlor on thyroid hormone receptor gene expression in the brain and behavior of Rana catesbeiana tadpoles

Abstract

The thyroid hormones (THs) including 3,5,3'-triiodothyronine (T3), are important regulators of growth and development of the brain in vertebrates. Previous studies showed that acetochlor, a widely used herbicide, accelerates T3 -induced frog tadpole metamorphosis and elevates the T3 -dependent accumulation of the mRNAs encoding the TH receptors, TRalpha and beta, in the tail. Here we show that acetochlor affects the expression of these TR isoforms in the brain of Rana catesbeiana tadpoles. Premetamorphic tadpoles exposed to 10 nM acetochlor with and without 100 nM T(3) for 4 days showed substantial increases in TRalpha and TRbeta transcript levels and significant decreases in the TRalpha/TRbeta ratios in their brains. This change in TR ratios is recapitulated with 10 nM acetochlor in R. catesbeiana tadpole brains during prometamorphosis, a period in which THs are endogenously produced. Tail fin biopsies revealed an elevation in TRalpha and beta mRNA levels compared to control animals when exposed to 1 and 10 nM acetochlor for 6 days. When subsequently reared in clean water for 59 days, no alterations in metamorphic hallmarks (forelimb emergence, mouth development, tail regression) were noted compared to the controls. Since alterations in TR ratios/levels may impact brain development, we tested the escape behavior in premetamorphic tadpoles exposed to 10 nM acetochlor for 4 days. We did not detect any statistically significant differences that would indicate that acetochlor affects escape behavior. However, since the gene expression data suggest that brain function may be affected, additional studies examining different behaviors upon acetochlor exposure at environmentally-relevant concentrations are warranted.