Characteristics of 3,5,3'-triiodothyronine (T3)-uptake system of tadpole red blood cells: effect of endocrine-disrupting chemicals on cellular T3 response.
We characterized the 3,5,3'-L-triiodothyronine (T3)- uptake system on the plasma membrane of Rana catesbeiana tadpole red blood cells (RBCs) in the presence of a variety of inhibitors and potentially competing amino acids. Saturable [125I]T3 uptake was inhibited by phloretin, monodansylcadaverine, bromosulfophthalein, sodium taurocholate and tryptophan. Saturable uptake obeyed simple Michaelis-Menten kinetics with an apparent Km of 110 nM and a Vmax of 2.5 pmol/min per 10(6) cells at 23 degrees C. These results suggested that a large proportion of T3 transported into RBCs was mediated by the aromatic amino acid transporter (System T)-linked transporter. To investigate the effect of endocrine-disrupting chemicals (EDCs) on [125I]T3 uptake, RBCs were incubated with [125I]T3 in the presence of each chemical. Among the test chemicals, di-n-butyl phthalate, n-butylbenzyl phthalate and the miticide, dicofol, were the most powerful inhibitors of [125I]T3 uptake, with an IC50 of 2.2 microM, which was one order of magnitude greater than that for T3 (IC50, 0.14 microM), and diethylstilbestrol and ethinylestradiol were modest inhibitors. Tributyltin accelerated saturable initial [125I]T3 uptake by 2-fold at 3.2 microM. When RBCs were cultured with 10 nM T3 at 25 degrees C for 2 days in the presence of monodansylcadaverine, ethinylestradiol, ioxynil or dicofol at the defined concentrations, these compounds inhibited significantly the induction of the thyroid hormone receptor alpha gene by T3. However, not all chemicals competed with T3 binding to the receptor at the same concentrations. Our results raise the possibility that the T3-uptake system on the plasma membrane of the tadpole RBCs could be a candidate target site for some EDCs and can modulate cellular T3 response.
Shimada
N
N
Department of Biology, Faculty of Science, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
Yamauchi
K
K
D000818:Animals; D001580:Benzopyrenes / Q000494:pharmacology; D002478:Cells, Cultured; D003864:Depression, Chemical; D004010:Dicofol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology; D007306:Insecticides / Q000494:pharmacology; D007457:Iodine Radioisotopes / Q000378:metabolism; D007814:Larva; D010795:Phthalic Acids / Q000494:pharmacology; D011892:Rana catesbeiana; D013974:Thyroxine / Q000494:pharmacology; D014220:Trialkyltin Compounds / Q000494:pharmacology; D014284:Triiodothyronine / Q000378:metabolism*
D001580:Benzopyrenes; D004967:Estrogens; D007306:Insecticides; D007457:Iodine Radioisotopes; D010795:Phthalic Acids; D014220:Trialkyltin Compounds; D014284:Triiodothyronine; C011559:tributyltin; C032279:phthalic acid; D013974:Thyroxine; D004010:Dicofol