Expression of sodium-iodide symporter mRNA in the thyroid gland of Xenopus laevis tadpoles: developmental expression, effects of antithyroidal compounds, and regulation by TSH.
The uptake of iodide represents the first step in thyroid hormone synthesis by thyroid follicular cells and is mediated by the sodium-iodide symporter (NIS). In mammals, expression of NIS is stimulated by TSH and transcription of the NIS gene involves regulation by the thyroid-specific transcription factors Pax8 and Nkx2.1. In this study, we examined the mRNA expression of NIS, Pax8 and Nkx2.1 in the thyroid gland of Xenopus laevis tadpoles by semi-quantitative reverse transcriptase (RT)-PCR. During spontaneous metamorphosis, NIS mRNA expression was low in premetamorphic tadpoles, increased throughout prometamorphosis, and peaked at climax stage 60. Analysis of TSH beta-subunit (TSHbeta) mRNA in the pituitary of the same tadpoles revealed a close temporal relationship in the expression of the two genes during metamorphosis, suggesting a regulatory role of TSH in the developmental expression of NIS. Treatment of tadpoles with goitrogenic compounds (sodium perchlorate and ethylenethiourea) increased TSHbeta mRNA expression (approximately twofold) and caused thyroid gland hyperplasia, confirming that feedback along the pituitary-thyroid axis was operative. Analysis of gene expression in the thyroid gland revealed that goitrogen treatment was correlated with increased expression of NIS mRNA (approximately 20-fold). In the thyroid gland organ culture experiments, bovine TSH (bTSH; 1 mU/ml) strongly induced NIS mRNA expression. This effect was mimicked by co-culture of thyroid glands with pituitaries from stage 58 tadpoles and by agents that increase intracellular cAMP (forskolin, dibutyryl-cAMP). In addition, it could be shown that thyroid glands of X. laevis tadpoles express Pax8 and Nkx2.1 mRNA in a developmentally regulated manner and that ex vivo treatment of thyroid glands with bTSH, forskolin, and cAMP analogs increased the expression of Pax8 and Nkx2.1 mRNA. This is the first report on developmental profiles and hormonal regulation of thyroid gland gene expression in amphibian tadpoles and, together, results reveal a critical role of TSH in the regulation of NIS mRNA expression in the thyroid gland of X. laevis tadpoles.
Opitz
Robert
R
Department of Inland Fisheries, Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Mueggelseedamm 310, D-12587 Berlin, Germany.
Trubiroha
Achim
A
Lorenz
Claudia
C
Lutz
Ilka
I
Hartmann
Sabine
S
Blank
Tobias
T
Braunbeck
Thomas
T
Kloas
Werner
W
D000818:Animals; D013956:Antithyroid Agents / Q000494:pharmacology; D002417:Cattle; D005031:Ethylenethiourea / Q000494:pharmacology; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D007814:Larva; D008675:Metamorphosis, Biological; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009924:Organ Culture Techniques; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D010472:Perchlorates / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017670:Sodium Compounds / Q000494:pharmacology; D027981:Symporters / Q000235:genetics*; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*; D000074482:Thyroid Nuclear Factor 1; D013972:Thyrotropin / Q000494:pharmacology; D037322:Thyrotropin, beta Subunit / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis / Q000378:metabolism*; D000099209:Sodium-Iodide Symporters
D013956:Antithyroid Agents; D005031:Ethylenethiourea; D009687:Nuclear Proteins; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors; D010472:Perchlorates; D012333:RNA, Messenger; D017670:Sodium Compounds; D027981:Symporters; D000074482:Thyroid Nuclear Factor 1; D013972:Thyrotropin; D037322:Thyrotropin, beta Subunit; D014157:Transcription Factors; D029867:Xenopus Proteins; D000099209:Sodium-Iodide Symporters; C499500:Pax8 protein, Xenopus; C000614411:nkx2-1 protein, Xenopus; C031068:sodium perchlorate