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Requirements of follicle structure for thyroid hormone synthesis; cytoskeletons and iodine metabolism in polarized monolayer cells on collagen gel and in double layered, follicle-forming cells.

Thyroid cells take up iodine and synthesize thyroid hormones. Thyroid cell polarity plays an important role in the uptake of iodine. However, we do not know whether polarity itself is enough for thyroid hormone synthesis or whether follicle structure is required for it. Using polarized monolayer porcine thyroid cells, cultured on collagen-coated filters, and double layered, follicle-forming cells, we analyzed the relationships of iodine metabolism and cell polarity (and follicle formation). We demonstrated that follicle structure was required for thyroid hormone synthesis. A quick-freezing and deep-etching method revealed the three-dimensional ultrastructures of cytoskeletons in the thyroid cells. On the collagen gel, the thyroid cells are reorganized into polarized monolayer cells; the basal cell membranes are in contact with the collagen gel and the apical ones face the culture medium. Actin microfilaments predominate under the apical cell membranes and intermediate filaments in the basal cytoplasm. The arrangement of these cytoskeletons determines the polarity of the cells. When the cells are cultured as double layers, follicle structures are reconstructed between the two monolayers. When apical cell membranes are in contact with other apical ones or when the cells are cultured as double layers, the cells are reorganized into follicles; the basal cell membranes are in contact with the collagen gel, and the apical ones face the follicle cavities. Actin microfilaments predominate at the apical cell membranes and intermediate filaments in the basal cytoplasm. Polarized thyroid cells transport iodine from the basal compartments to the apical ones, but cannot organify iodine into thyroid hormones. However, follicle-forming cells, cultured as double layers, take up iodine and organify it into thyroid hormones. Polarity is important for iodine uptake, and follicle structure is required for thyroid hormone synthesis.

Animals↗

Prevalence of thyroid hormone autoantibodies in healthy subjects.

OBJECTIVE: The prevalence of thyroid hormone autoantibodies in patients with thyroid disorders has been well described, although the results have been variable. However, the prevalence of thyroid hormone autoantibodies in apparently healthy subjects remains unknown and its determination was the main objective of this study. SUBJECTS AND METHODS: Sera obtained from 880 healthy subjects (365 men and 515 women) were examined for thyroid hormone autoantibodies by immune precipitation using radiolabelled thyroid hormones or thyroid hormone analogues. RESULTS: Anti-triiodothyronine (T3) and anti-thyroxine (T4) antibodies were detected in none (0%) and in 3 (0.34%), respectively, of the 880 individuals studied using radiolabelled thyroid hormones. Similar tests in 385 healthy subjects using radiolabelled thyroid hormone analogues (polyaminocarboxy T3 and T4) showed the presence of anti-T3 and anti-T4 antibodies in 3 (0.78%) and in 4 (1.04%), respectively. None of the subjects had both anti-T3 and anti-T4 antibodies. Thus, the prevalence of anti-T3 or anti-T4 antibodies among healthy population was concluded to be as high as 1.8%. Radiolabelled thyroid hormone analogue binding to purified immunoglobulin G (IgG), with or without the addition of an excess of unlabelled thyroid hormones, confirmed specific binding of thyroid hormones to the thyroid hormone autoantibody-positive IgGs. The presence and class specificity of thyroid hormone autoantibodies was confirmed by the Ouchterlony immunodiffusion/autoradiography method, which showed biphenotypic heavy chain (IgG and IgA) and kappa (kappa) light chain specificities. Serum concentrations of free thyroid hormones and TSH in thyroid hormone autoantibody-positive sera were within the normal range. CONCLUSIONS: These results indicate that the occurrence of thyroid hormone autoantibodies among the healthy Japanese population is fairly common. However, interference of thyroid hormone autoantibodies in the radioimmunoassay of free thyroid hormones is exceptional.

Adult↗

Thyroid hormones, gonadal and adrenocortical steroids and the function of the islets of Langerhans.

Thyroid hormones, gonadal and adrenocortical steroids, are glucoregulatory hormones. Thyroid hormones increase the provision of glucose to meet the enhanced energy demands which they impose. Glucose tolerance is decreased, associated with increased hepatic glucose production, although the glucose-raising effects of thyroid hormones are partially offset by an increased rate of glucose utilization especially in the postabsorptive state. The insulin secretory capacity of the pancreatic B cells is reduced by an excess of thyroid hormones, and the onset of diabetes may be hastened as pancreatic insulin reserves are depleted. Natural estrogens can improve glucose tolerance through a beta-cytotropic effect and enhanced insulin sensitivity. Progesterone may produce similar effects in the absence of estrogens, but progestins appear to antagonize the effects of estrogens. Testosterone exerts only marginal effects on glucose tolerance. Glucocorticoids decrease glucose tolerance by increased hepatic glucose production and impaired peripheral glucose utilization. Glucocorticoids reduce insulin sensitivity and responsiveness in peripheral tissues. However, the diabetogenic influence of glucocorticoid excess is partly compensated by a beta-cytotropic effect and a condition of diabetes develops when the functional reserve of the endocrine pancreas becomes limiting.

Adrenal Cortex Hormones↗

[Thyroid transcription factors involved in the thyroid hormones biosynthesis].

The thyroid hormone biosynthesis is dependent on the level of the expression and the correct action of the basic proteins involved in this process: TSH receptor (TSH-R), sodium-iodine symporter (NIS), thyroglobulin (Tg), thyroid peroxidase (TPO) and iodothyronine deodinases type 1 (D1) and type 2 (D2). Transcription of the genes coding these proteins is regulated by TSH via cAMP cascade. Despite the thyroid-specific transcription factors are necessary for the transcription regulation of TSH-R, NIS, TG, TPO, DIO1 and DIO2 genes. Three thyroid transcription factors Titf1/Nkx2-1, Foxel and Pax8 seem to be essential for thyroid development, growth and function. Disturbances of the expression of these transcription factors and their co-operation could be responsible for the congenital thyroid gland disorders as well as in thyroid cancerogenesis.

Animals↗

Splicing of juvenile and adult tau mRNA variants is regulated by thyroid hormone.

The effect of thyroid hormone on the expression of tau transcripts was studied during postnatal brain development. The level of tau mRNA was only slightly changed postnatally in the cerebral hemispheres of hypothyroid rats, whereas the level of tau mRNA in the cerebellum was maintained at a higher level than in the euthyroid controls. As shown by in situ hybridization studies, such an alteration in tau mRNA expression can be ascribed to an effect of thyroid hormone on the rate of migration of the granule cells in the cerebellum; that tau mRNAs remain high in the cerebellum as long as the granule cells are migrating correlates with the observation that hypothyroidism slows the rate of migration of granule cells. RNase protection assays also showed that thyroid hormone deficiency delays the transition between the immature and mature tau transcripts in both brain regions. Thus, one of the effects of thyroid hormone is to regulate the splicing mechanism that allows replacement of the juvenile tau variants by the adult entities during neuronal differentiation.

Animals↗

Chicken ovalbumin upstream promoter transcription factor (COUP-TF) dimers bind to different GGTCA response elements, allowing COUP-TF to repress hormonal induction of the vitamin D3, thyroid hormone, and retinoic acid receptors.

Alignment of natural chicken ovalbumin upstream promoter transcription factor (COUP-TF) response elements shows that, in addition to the predominant direct repeat of the GGTCA motif with a 2-bp spacing, there are other functional COUP elements with variations in the GGTCA orientation and spacing. We systematically analyzed the binding of in vitro-synthesized COUP-TFs and showed that COUP-TF is capable of binding to oligonucleotides containing both direct repeats and palindromes and with different spacings of the GGTCA repeats. Subsequently, we analyzed four possible mechanisms proposed to explain how COUP-TF could bind to these spatial variations of the GGTCA repeat. We demonstrated that the functional DNA-binding form of COUP-TF is a dimer which requires two GGTCA half-sites to bind DNA. We demonstrated that the COUP-TF dimer undergoes a remarkable structural adaptation to accommodate binding to these spatial variants of the GGTCA repeats. A functional consequence of the promiscuous DNA binding of COUP-TF is its ability to down-regulate hormonal induction of target gene expression by other members of the steroid-thyroid hormone receptor superfamily such as the vitamin D3, thyroid hormone, and retinoic acid receptors. Our data indicate that COUP-TF may have an important role in hormonal regulation of gene expression by these receptors.

Animals↗

[Cellular mechanisms of thyroid hormone action].

KEY POINTS: Thyroid hormones affect cardiac myocytes as well as the smooth muscle and endothelial cells of the vascular wall. Free 3,53'-L-triiodothyronine (FT3) and its specific nuclear receptor modulate the transcription of various proteins, principally those involved in the myocyte contractile apparatus (myosin heavy chains), and the regulation of intracellular calcium flux (sarcoplasmic reticulum Ca2+ATPase). Thyroid hormones also have non-genomic effects that work rapidly, complement the effects described above, and are related to alterations in the properties of many channels and membrane receptors, especially in the sinoatrial mode. Thyroid hormones also affect the smooth muscle and endothelial cells of the vascular walls and reduce systemic vascular resistance. These effects on cardiac and vascular cells globally explain the cardiac manifestations (especially the inotropic and chronotropic effects) observed during dysthyroidism, particularly in hyperthyroidism where they are often in the forefront (positive inotropic and chronotropic effects).

Animals↗

Autoimmunity against thyroid hormones.

The presence of thyroid hormone autoantibodies (THAA) is a common phenomenon. More than 270 cases have been reported by the end of 1993 involving not only thyroidal but also nonthyroidal disorders. Clinically, THAA in a patient's serum produces variation in thyroid hormone metabolism and, in particular, may interfere with the radioimmunoassay (RIA) results of total or free thyroid hormone measurements, which can cause unusually high or low values of the hormones depending on the B/F separation method used. This in vitro interference can give clinicians confusing information about the patient's thyroid state. As a result, the patient may receive inappropriate treatment from physicians who are unaware of this disorder. The presence of THAA has been reported not only in humans but also in dogs, chickens, and rats. In this review article, clinical features of THAA and the mechanism of autoantibody production are discussed.

Adolescent↗

Re-evaluation of thyroid hormone status after long-term hormone therapy.

Once thyroid hormone is prescribed, it is usually continued throughout a persons's lifetime. It is therefore important that the diagnosis be based on the most definitive test available. In this study with a primary care population of 6,000 individuals, 24 patients were found to carry the diagnosis of hypothyroidism. A careful view of the records showed that the diagnosis was inadequately documented by current standards in 20 of these 24 individuals. A 21-day hormone withdrawal test was done in ten of the patients in whom the diagnosis was suspected, and 60 percent were found to be euthyroid. Although the number of patients was small, this does encourage the speculation that many patients diagnosed in the past as hypothyroid may in fact be euthyroid. The medical as well as economic impact of this high rate of misdiagnosis becomes apparent when the one contemplates the 15,000,000 prescriptions for thyroid hormone written annually in the United States.

Adult↗

Cellular binding proteins of thyroid hormones.

Cellular binding proteins of thyroid hormones are present in the cell nucleus, cytosol, cell membrane, and mitochondria. While nuclear binding is proven to mediate hormone action, the exact roles of the other binding sites remain to be established. Nuclear receptor associates with DNA, core histone, and nuclear matrix and preferentially distributes in transcriptionally active chromatin due to interaction with H1 histone. Of particular importance is the binding of nuclear receptor to specific DNA sequences of target genes, termed thyroid-responsive elements. The binding is stabilized by non-receptor nuclear protein. Upon binding thyroid hormone, nuclear receptor is activated through alterations in the steric configuration, leading to changes in the rate of transcription of the target genes. Multiple nuclear receptor forms exist with likely distinct functional roles. Cytosolic thyroid hormone binding proteins are also heterogeneous. One form is under the control of cell metabolism (NADP and NADPH) and it may have a role in transport of the hormone to mitochondria and nucleus. Membrane-linked thyroid hormone binding proteins may have dual functional roles: one is to mediate hormone action and the other is to support active uptake of hormones by cells. Mitochondrial function may be regulated by thyroid hormone through mitochondrial binding sites in cooperation with nuclear receptor-mediated pathway. Further studies are required to elucidate the exact functional roles of non nuclear thyroid hormone binding proteins.

Animals↗

Developing cholinergic basal forebrain neurons are sensitive to thyroid hormone.

The influence of thyroid hormone on the development of cholinergic neurons in nucleus basalis was assessed in hyperthyroid, hypothyroid, and euthyroid rats by use of CAT immunohistochemistry and single-section Golgi-impregnation histology. Animals were made either hyperthyroid by daily injections of 1.0 micrograms/gm body weight triiodothyronine starting at postnatal day (P) 3 or hypothyroid by providing 0.4% propylthiouracil in the diet of dams from P2. Compared to developing control rats, increased exposure to thyroid hormone resulted in accelerated expression of CAT in nucleus basalis neurons. Overshoot in cell body size, a normal developmental phenomenon of cholinergic neurons in the basal nuclear complex, occurred earlier in hyperthyroid brains and was of a greater magnitude than in controls. Furthermore, increased numbers of primary dendrites and dendritic branchpoints accompanied by dendritic and perisomal filopodia-like structures were observed for nucleus basalis neurons in hyperthyroid rats. These dendritic changes persisted throughout the second postnatal month. After the fifth postnatal week, cell body sizes of these hyperthyroid CAT-positive neurons began to decrease and by P50 were significantly less than controls or similarly treated animals at earlier ages. By P64, the number of cholinergic neurons in nucleus basalis was appreciably less than in age-matched controls. Hypothyroidism resulted in a delay of normal CAT expression that persisted throughout the third postnatal week. After this time, CAT staining increased until normal immunoreactivity was attained in cell bodies, fibers, and terminal regions by P35. A deficit in thyroid hormone during development prevented overshoot in perikaryal size and resulted in diminished cross-sectional areas throughout the cholinergic nucleus basalis at all ages examined. Hypothyroidism also prevented the normal overproduction of dendrites in those cells and produced stunted dendritic trees at all ages examined. These morphological abnormalities persisted throughout the second postnatal month. The effects of thyroid hormone on cholinergic projection neurons in the rat brain appeared relatively selective for cells in the basal nuclear complex because neither hypothyroid nor hyperthyroid treatment produced changes in the cell body areas of the phenotypically similar CAT-positive neurons of the pontomesencephalotegmental complex.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Studies on the mechanism by which thyroid hormones enhance alpha-lactalbumin activity in explants from mouse mammary glands.

Addition of 3,5,3'-triiodo-L-thyronine to cultures of mammary gland explants in serumfree medium containing insulin, hydrocortisone and prolactin results in a 3 to 5-fold increase in the activity of the milk-protein alpha-lactalbumin over that seen in the presence of the latter three hormones alone. The thyroid hormone does not act by substituting for any of the other hormones. It need not be present throughout the culture period but can act if added along with prolactin after insulin and hydrocortisone have induced formation of the rough endoplasmic reticulum. Delayed addition of the thyroid hormone also results in further stimulation of cells already responding maximally to insulin, hydrocortisone and prolactin. These effects of triiodothyronine are not blocked by progesterone at 1 microng per ml. They are, however, blocked by the addition of inhibitors of RNA (actinomycin D) or protein (cycloheximide or puromycin) synthesis, suggesting that the thyroid hormone increases the synthesis of the alpha-lactalbumin molecule itself. The thyroid hormone appears to act by altering the responsiveness of the mammary gland explants to prolactin, but not to insulin or hydrocortisone. In the presence of 10-9M triiodothyronine, enhanced alpha-lactalbumin activity is consistently obtained at prolactin concentrations as low as 4.5 x 10(-12)M whereas, in the absence of the thyroid hormone, ten times more prolactin (4.5 x 10(-11)M) is needed to obtain an increase in alpha-lactalbumin activity.

Animals↗

Molecular cloning and properties of a full-length putative thyroid hormone receptor coactivator.

Thyroid hormone receptors (TRs) are ligand-dependent transcription factors that regulate target gene transcription. The conserved carboxy-terminal region of the ligand-binding domain (AF-2) has been thought to play a critical role in mediating ligand-dependent transactivation by the interaction with coactivator(s). Using bacterially-expressed TR as a probe, far-Western-based expression cDNA library screening identified cDNAs that encode, in part, the recently reported partial steroid receptor coactivator-1 (SRC-1) sequence. Additional work, including 5' RACE, has characterized a full-length cDNA that encodes a approximately 160 kD protein as a putative thyroid hormone receptor coactivator (F-SRC-1). In vitro binding studies show that F-SRC-1 binds to a variety of nuclear hormone receptors in a ligand-dependent manner, along with TBP and TFIIB, suggesting that F-SRC-1 may play a role as a bridging molecule between nuclear hormone receptors and general transcription factors. Interestingly, AF-2 mutants also retain ligand-dependent interaction with F-SRC-1. Although F-SRC-1 recognizes the ligand-induced conformational changes of nuclear hormone receptors, our observations suggest that F-SRC-1 may bind directly with subregion(s) in nuclear hormone receptors other than the AF-2 region.

Amino Acid Sequence↗

Genetic analysis of thyroid hormone receptors in development and disease.

Thyroid hormone (T3) fulfills diverse functions in vertebrate development and physiology. These functions are thought to be mediated by two genes encoding the related T3 receptors. TR alpha and TR beta. The use of homologous recombination in embryonic stem cells to generate defined, single-gene mutations provides a powerful means to investigate the individual functions of TR alpha and TR beta in mice. We have shown that targeted inactivation of the TR beta gene results in goiter and elevated levels of thyroid hormone. Thyroid stimulating hormone (TSH), which is released by pituitary thyrotropes and is normally suppressed by increased levels of thyroid hormone, was present at elevated levels in homozygous mutant (Thrb-/-) mice. These findings suggest a unique role for TR beta that cannot be substituted by TR alpha in the T3-dependent feedback regulation of TSH transcription. Thrb-/- mice provide a recessive model for the human syndrome of resistance to thyroid hormone (RTH). Typically, RTH is associated with dominant mutations in TR beta. It is unknown whether TR alpha, TR beta, or other receptors are targets for inhibition in dominant RTH; however, the analysis of Thrb-/- mice suggests that antagonism of TR beta-mediated pathways underlies the disorder of the pituitary-thyroid axis. Thrb-/- mice also display defective maturation of auditory function, demonstrating that TR beta is essential for the development of hearing. Interestingly, hearing defects are generally absent in dominant RTH, indicating that in the auditory system, a dominant TR beta mutant cannot mimic the defect caused by loss of TR beta. This suggests the existence of tissue-specific mechanisms that modulate the activity of TR beta. These results define in vivo functions for TR beta and indicate that specificity in T3 signaling is conferred by distinct receptor genes.

Animals↗

A comparison of short-term changes in health-related quality of life in thyroid carcinoma patients undergoing diagnostic evaluation with recombinant human thyrotropin compared with thyroid hormone withdrawal.

CONTEXT: Thyroid carcinoma requires lifelong monitoring with serum thyroglobulin, radioactive iodine whole body scanning, and other imaging modalities. Levothyroxine (L-T4) withdrawal for thyroglobulin measurement and whole body scanning increases these tests' sensitivities but causes hypothyroidism. Recombinant human TSH (rhTSH) enables testing without L-T4 withdrawal. OBJECTIVE: Our objective was to examine the impact of short-term hypothyroidism on the health-related quality of life (HRQOL) of patients after rhTSH vs. L-T4 withdrawal. DESIGN, SETTING, AND PATIENTS: In this multicenter study, the SF-36 Health Survey was administered to 228 patients at three time points: on L-T4, after rhTSH, and after L-T4 withdrawal. INTERVENTIONS: INTERVENTIONS included administration of rhTSH on L-T4 and withdrawal from thyroid hormone. MAIN OUTCOME MEASURES: Mean SF-36 scores were compared during the two interventions and with the U.S. general population and patients with heart failure, depression, and migraine headache. RESULTS: Patients had SF-36 scores at or above the norm for the general U.S. population in six of eight domains at baseline on L-T4 and in seven of eight domains after rhTSH. Patients' scores declined significantly in all eight domains after L-T4 withdrawal when compared with the other two periods (P < 0.0001). Patients' HRQOL scores while on L-T4 and after rhTSH were at or above those for patients with heart failure, depression, and migraine in all eight domains. After L-T4 withdrawal, patients' HRQOL scores were significantly below congestive heart failure, depression, and migraine headache norms in six, three, and six of the eight domains, respectively. CONCLUSIONS: Short-term hypothyroidism after L-T4 withdrawal is associated with a significant decline in quality of life that is abrogated by rhTSH use.

Female↗

Characterization of the uridine diphosphate-glucuronosyltransferase-catalyzing thyroid hormone glucuronidation in man.

Increased thyroid hormone glucuronidation in rats caused by exposure to xenobiotics has stimulated a search for the individual uridine diphosphate-glucuronosyltransferases (UGTs) catalyzing this reaction in rats and man. Microsomal preparations from Crigler-Najjar liver, normal human liver, and kidney have been used to try to identify the UGT isoforms responsible for glucuronidation of the thyroid hormones. The predominant thyroid hormone released from the thyroid gland, T4, and the inactive rT3 are glucuronidated by cloned expressed bilirubin UGT1A1 and also phenol UGT1A9. Results from Crigler-Najjar microsomal samples indicate that UGT1A1 is the main contributor to thyroid hormone glucuronidation in the liver, with rT3 being the preferential substrate. In kidney microsomes thyroid hormone glucuronidation is more complex, suggesting that more than just the UGT1A9 isoform may be involved. Bioactive T3 is not significantly glucuronidated by these isoforms and other UGTs, and sulfotransferases may be involved.

Animals↗

Mice with a mutation in the thyroid hormone receptor beta gene spontaneously develop thyroid carcinoma: a mouse model of thyroid carcinogenesis.

The molecular genetic basis of thyroid carcinogenesis is not well understood. Most of the existing models of thyroid cancer only rarely show metastases, and this has limited progress in the understanding of the molecular events in thyroid cancer invasion and metastasis. We have recently generated a mutant mouse by introducing a dominant negative mutant thyroid hormone nuclear receptor gene, TRbetaPV, into the TRbeta gene locus. In this TRbetaPV mouse, the regulation of the thyroid-pituitary axis is disrupted, leading to a mouse with high levels of circulating thyroid-stimulating hormone and extensive hyperplasia of follicular epithelium within the thyroid. As TRbeta(PV/PV) mice, but not TRbeta(PV/+) mice, aged, metastatic thyroid carcinoma developed. Histologic evaluation of thyroids of 5-14-month-old mice showed capsular invasion (91%), vascular invasion (74%), anaplasia (35%), and metastasis to the lung and heart (30%). Previous models of thyroid cancer have focused on genes that control initial carcinogenesis, but this model provides an unusual opportunity to study the alterations in gene regulation that occur with clinically relevant changes during progression and metastasis in a predictable fashion.

Anaplasia↗

Immunohistochemistry of thyroid hormones as a functional parameter in thyroid pathology.

The authors study by means of immunoperoxidase method the pattern of thyroglobulin, triiodothyronine and thyroxine distribution in 58 cases of thyroid disorders: 15 euthyroid goiters, 10 Graves' disease, 7 Hashimoto's thyroiditis, 11 folliculo-papillary carcinomas (6 primary tumors and 5 lymph node metastases), 8 follicular carcinomas, 4 anaplastic carcinomas and 3 medullary carcinomas. Thyroglobulin, triiodothyronine and thyroxine were present in most of the thyroid disorders, excepting anaplastic and medullary carcinomas. Thyroglobulin and thyroxine were localized both in the follicular epithelium and in the colloid, whereas triiodothyronine was present especially in the follicular cells. The thyroid hormones distribution in benign lesions is rather similar. In carcinomas, the pattern of thyroglobulin, triiodothyronine and thyroxine is more heterogeneous, but generally the triiodothyronine distribution is similar to that of thyroglobulin. In some carcinomas, triiodothyronine and thyroxine showed a weak or negative immunostaining. The immunoperoxidase method is a valuable tool in the study of functional disturbances in the thyroid pathology and in the diagnosis of thyroid carcinoma metastases as well. Positive thyroid hormones staining clearly indicates the thyroid origin of metastases.

Histocytochemistry↗