Role Of TSH Research Articles

Expression of the Human TSH Receptor in a Human Thyroid Carcinoma Cell Line That Lacks an Endogenous TSH Receptor: Growth Inhibition by cAMP

The role of TSH and its receptor in controlling growth of thyroid carcinomas is far from well understood. In order to study this subject further we established a new human thyroid carcinoma cell line. We transfected human thyroid carcinoma cells lacking an endogenous TSH receptor with the human TSH receptor cDNA. Transfected cells, designated HTC-TSHr, expressed the TSH receptor mRNA and synthesized a functional TSH receptor with a TSH binding affinity in the order of magnitude of normal thyroid cells. In response to TSH stimulation HTC-TSHr cells accumulated cAMP, indicating a functional TSH receptor-adenylate cyclase system. However, HTC-TSHr cells did not concentrate iodide and lacked thyroglobulin immunoreactivity, although they did express low amounts of thyroglobulin mRNA. Proliferation of HTC-TSHr cells was inhibited by dibutyryl-cAMP and forskolin and also by TSH via the re-expressed TSH receptor.

Thyroid and growth factors

Although TSH is the major regulator of thyroid growth and function, the role of TSH in thyroid cell growth in vitro is controversial. Recent studies have shown that a number of growth factors, cytokines, vitamins, hormones and other reagents including iodide, lithium or retinoids modulate proliferation of cultured thyroid cells. Insulin-like growth factors (IGFs) stimulate growth of thyroid cells from a variety of species and also enhance expression of differentiated functions stimulated by TSH. Epidermal growth factor (EGF), fibroblast growth factor (FGF), and Transforming growth factor (TGF)-alpha are also mitogens to thyroid cells, but they are inhibitory to differentiated functions. In contrast, TGF-beta is a potent inhibitor to thyroid cell growth and functions. Indirect evidence suggests that these growth factors are involved in growth regulation of normal and neoplastic thyroid cell growth in autocrine/paracrine manner.

Thyrotropin Stimulates the Expression of an Acidic Ribosomal Protein, PO, Messenger Ribonucleic Acid in Cultured Rat Thyroid (FRTL) Cells

Acidic ribosomal proteins, P0, P1, and P2 in eukaryotic 60S subunits play an important role in polypeptide chain elongation during the translational step. To investigate the role of TSH in protein synthesis in the thyroid, we examined the effect of TSH on ribosomal P-protein biogenesis in FRTL cells. First, we investigated the influence of TSH on P0-protein gene expression. RNA slot blot hybridization revealed that the effect of TSH on P0-protein mRNA accumulation in the quiescent FRTL cells was time- and dose-related. This stimulatory effect of TSH was mimicked by (Bu)2cAMP. Nuclear run-off transcription assays revealed that TSH increased the transcriptional activity of the P0-protein mRNA without an increase in beta-actin transcriptional activity. On the contrary, the stability of P0-protein mRNA decreased after the addition of TSH. Cycloheximide markedly inhibited TSH-induced P0-protein mRNA accumulation in FRTL cells. Second, using a synthetic oligonucleotide probe, we have shown that TSH also increased P2-protein mRNA levels in FRTL cells. Furthermore, immunodetection of P-proteins using anti-P-protein antibody showed that TSH significantly increased the amount of P-proteins in the cells. These results suggest that TSH can increase the amount of acidic ribosomal P-proteins at least in part through an increase in the level of P-protein gene transcripts. This effect occurs via a transcriptional mechanism and requires ongoing protein synthesis. Thus, TSH might play an important role in ribosome biogenesis in FRTL cells.

Compensatory changes in thyroid blood flow are only partially mediated by thyrotropin

It has been shown that the compensatory growth of the thyroid gland and the compensatory increase in hormone secretion that occur after hemithyroidectomy are preceded by a dramatic increase in thyroid blood flow (BF). These alterations in the thyroid remnant may be due to the concomitant increase in plasma thyrotropin (TSH) concentrations. It has been suggested, however, that the compensatory thyroid growth may also involve a neural reflex. In this study we have investigated the role of TSH in mediating the compensatory alterations in thyroid BF and mass after subtotal thyroidectomy. Male Sprague-Dawley rats were anesthetized with ether for surgical or sham hemithyroidectomy. One-half of the hemithyroidectomized rats (HTX) received no further treatment; in the other one-half of the HTX rats (Clamp), plasma TSH levels were maintained at levels comparable with those in sham-operated animals by initiating constant thyroid hormone replacement beginning at the time of hemithyroidectomy. Plasma samples for TSH, 3,5,3'-triiodothyronine, and thyroxine radioimmunoassays were obtained 2, 7, 14, and 21 days after surgery. Thyroid BF was determined at 1, 2, and 3 wk after surgery by the reference sample version of the radioactive microsphere technique (141Ce, 15 microns diameter). Plasma TSH levels and thyroid lobe weight were significantly elevated in HTX rats but not in Clamp rats. Thyroid BF was markedly increased in HTX rats. Thyroid BF was also significantly increased in Clamp rats despite the suppression of the rise in plasma TSH concentration, but this increase was less than that in HTX rats. Neither hemithyroidectomy nor Clamp treatments had any effect on arterial blood pressure or BF to other tissues (e.g., kidney).(ABSTRACT TRUNCATED AT 250 WORDS)

Insulin and insulin-like growth factor (IGF I) modulate the effects of bTSH on 3H-thymidine incorporation in human thyroid cells in primary culture.

The role of TSH in thyroid cell growth and the pathogenesis of goiter has become a matter of recent debate, since many investigators have failed to demonstrate a growth-promoting effect of TSH in human thyroid cells in culture. While those studies have focused on the action of TSH in human thyroid cells, the influence of assay conditions and cofactors has received scant attention. In the present study, we have therefore undertaken to elucidate the effects of insulin and insulin-like growth factor (IGF I) on 3H-thymidine uptake in human thyroid cells, particularly with respect to their relation to the actions of bTSH. We could demonstrate a considerable, dose-dependent stimulation of 3H-thymidine incorporation in the cells by bTSH that was dependent on the presence of insulin or IGF I; bTSH alone was ineffective in that respect. The concentrations of insulin and IGF I required to facilitate the TSH response were of a magnitude at which both peptides were totally ineffective by themselves. At concentrations of insulin or IGF I that produced a maximum stimulation of 3H-thymidine incorporation, the addition of bTSH did result in a slight decrease rather than a further increase of that stimulation. We conclude from these findings, first, that TSH appears to be a growth factor for human thyroid cells under the conditions described, and, second, the effects of TSH on thyroid cell proliferation are under the control of cofactors like insulin and IGF I.

Thyrotropin Binding to Rat Thyroid Membranes: Reduced Capacity Associated with Goitrogenesis*

To investigate a possible role of TSH in the regulation of its own receptor, a sensitive assay of [125I]TSH tropin binding to rat thyroid membranes was used. With 1 mM MgCl2 in the buffer, Scatchard analysis of displacement of TSH gave a curvilinear plot with a high affinity, low capacity (K1, 3.4 nM; Q1, 3.1 pmol/microgram) and a low affinity, high capacity binding site (K2, 0.54 microM; Q2, 1.2 X 0.1 nmol/microgram). Feeding rats propylthiouracil led to a decrease in [125I]TSH binding (expressed either per U protein or per wet wt of tissue) that was related to the duration of treatment. Evaluation by Scatchard analysis showed that this was due to a loss of binding sites, e.g. a 50-60% decrease after 1 month of propylthiouracil treatment; affinity was actually slightly increased in the goitrous tissue. This change in the number of TSH-binding sites was readily reversed in association with the suppression of TSH in vivo either by injections of T3 for 3 days or more by feeding a normal diet for 1 month. Thus, the data are compatible with TSH, that is in high concentration in the serum of rats fed propylthiouracil, exerting a downregulatory influence on its own receptors.

Regulation of thyroidal deiodinase activity.

In previous work with human multinodular goiters we had observed widely differing deiodinase activities in different tissue samples of the same thyroid. Since all areas of a single gland must be exposed to an identical plasma TSH level, the findings suggested that TSH is not the only factor regulating deiodinase activity in a goitrous gland. The present investigation was designed to define the relative role of TSH and non-TSH-dependent factors in regulating deiodinase activity of the rat thyroid. After 7 days of goitrogen treatment, the deiodinase activity per g thyroid and per mg DNA was greatly increased with perchlorate (C1O4-) and 6-propyl-2- thiouracil plus C1O4-, and only slightly increased with propylthiouracil alone and with a low iodine diet. Despite the continuous rise of serum TSH with further treatment, deiodinase activity per mg DNA declined to well below peak levels after 1 more week and remained at this only slightly higher than normal value for many weeks irrespective of the goitrogenic re...

Role of TSH in the Changes in Thyroidal Metabolism of [35S]Methimazole in Phenobarbital and Thyroxine-Treated Rats

The effect of phenobarbital (PB) and/or thyroxine on the thyroidal accumulation and oxidation of [35S]methimazole (MMI) and serum TSH levels was studied in rats. PB treatment increased the accumulation of MMI and the serum TSH levels, but concurrent administration of T4 reversed these effects. It was concluded that increased TSH secretion in PB-treated animals was likely to be the major mechanism involved in the increased MMI accumulation. PB also increased the intrathyroidal oxidation of MMI to sulphate. However, in contrast to the PB effect on accumulation, concurrent T4 administration only partially reversed the effect on oxidation. The results suggested that the increased oxidation of MMI in PB-treated animals was due to a direct effect of PB or possibly a combination of this direct effect and the indirect TSH effect. Possible mechanisms postulated for a direct effect were thyroidal microsomal enzyme induction and/or changes in thyroidal protein binding of MMI.

Compensatory thyroid hypertrophy after hemithyroidectomy in rats.

Thyroid enlargement occurs in association with a variety of circumstances characterized by an impaired capacity of the gland to secrete adequate amounts of hormone. To elucidate the factors responsible for such compensatory thyroid growth, particularly the role of TSH, we have observed the response of the serum TSH, T3 and T4 concentrations following hemithyroidectomy in the rat, and have attempted to correlate changes in these functions with changes in the weight and histology of the thyroid remnant. Hemithyroidectomy was performed in male Sprague-Dawley rats weighing 150 to 370 g, sham-operated animals serving as controls. As compared to findings in sham-operated animals, serum T4 concentrations declined promptly after hemithyroidectomy. In Experiment I serum T4 concentrations remained low for about 10 days and then returned to initial values. In Experiment II serum T4 concentrations remained lower than initial T4 values or values found in sham-operated animals until 34 days after hemithyroidectomy. Serum T3 concentrations were not significantly altered after hemithyroidectomy in either group but tended to be lower in the hemithyroidectomized animals. Serum TSH concentrations increased within 3 days after hemithyroidectomy and, for as long as 21 weeks, remained at values higher than those present preoperatively or those seen in sham-operated animals. Thyroid lobe weight increased following removal of the contralateral lobe and this increase was also sustained throughout the duration of the experiments. Biochemical and histological observations indicated that enlargement of the residual lobe was due to hypertrophy rather than hyperplasia.

Aspects ultrastructuraux et synthèse de la thyroglobouline dans les cultures de thyroides avant et après transplantation chez des rats hypophysectomisés

Two aspects concerning the study of thyroglobulin synthesis and of the ultrastructure of thyroid glands in organotypic cultures have been studied: (1) the chronology of ultrastructural alterations in thyroid cultures leading to the loss of thyroglobulin synthesis; and (2) the role of TSH in the re-establishment of the characteristic features in the cultured cells following transplantation. Alterations of the endoplasmic reticulum start on the very first day of culture. The ribosomes detach themselves from the membranes which then disappear. On the third and fourth day, significant alterations are observed on mitochondria and nuclei. The lysosomes disappear, while the Golgi apparatus is hardly affected. All the above modifications are reversible, since it is possible to re-establish the characteristic features of the endoplasmic reticulum and the synthesis of thyroglobulin after transplantation of the cultured glands into normal animals. The reappearance of phenotypic traits of the thyroid cells can also be obtained after transplantation into hypophysectomized rats. However, the level of thyroglobulin synthesis in these transplants, which is similar to that in the host gland, is lower than in grafts into normal animals. This results shows that TSH is not required for differentiation and for manifestation of the phenotypic traits of the thyroid gland. It seems therefore that the role of TSH is not to stimulate differentiation, but to maintain and regulate the level of specific processes in the already differentiated thyroid cell.