Thyroid-stimulating hormone receptor and its role in Graves' disease

The thyroid-stimulating hormone (TSH, or thyrotropin) receptor (TSHR) mediates the activating action of TSH to the thyroid gland, resulting in the growth and proliferation of thyrocytes and thyroid hormone production. In Graves' disease, thyroid-stimulating autoantibodies can mimic TSH action and stimulate thyroid cells. This leads to hyperthyroidism and abnormal overproduction of thyroid hormone. TSHR-antibodies-binding epitopes on the receptor molecule are well studied. Mechanism of TSHR-autoantibodies production is more or less clear but a susceptibility gene, which is linked to their production, is still unknown. Genetic studies show no linkage between the TSHR gene and Graves' disease. Among three common polymorphisms in the TSHR gene, only the D727E germline polymorphism in the cytoplasmic tail of the receptor showed an association with the disease, and this association is weak. The absence of a strong genetic effect of the TSHR polymorphisms in such a common and complex disorder as Graves' disease may be explained by a high degree of evolutionary conservation in TSHR. This can be shown by naturally existing germline and somatic mutations in the TSHR gene that cause various types of nonautoimmune and hereditary thyroid disease.     

An observation that illustrates most T cell receptor structure-function relationships

The Standard model of the T cell receptor (TCR) structure-function relationships is based on an analogy with the B cell receptor. Here a single observation is analyzed to show why this appears to be untenable. The Standard model cannot account for allele-specific recognition of theMHC-encoded presenter of peptide (R) by the TCR nor can it adequately explain alloreactivity. The competing framework is based on the assumptions that (1) single V-domains recognize the alleles of R, (2) restrictive reactivity is peptide specific, whereas alloreactivity is peptide unspecific, and (3) the TCR is born in two conformations, which display reciprocal behaviors (see text). In any case, whatever position one takes regarding these two models, competing conceptualizations are of crucial value in guiding experimentation, not to mention creative thinking.     

Superantigens: structure-function relationships

Superantigens are a class of highly potent immuno-stimulatory molecules produced by Staphylococcus aureus and Streptococcus pyogenes. These toxins possess the unique ability to interact simultaneously with MHC class II molecules and T-cell receptors, forming a trimolecular complex that induces profound T-cell proliferation. The resultant massive cytokine release causes epithelial damage and leads to capillary leak and hypotension. The staphylococcal superantigens are designated staphylococcal enterotoxins A, B, C (and antigenic variants), D, E, and the recently discovered enterotoxins G to Q, and toxic shock syndrome toxin-1. The streptococcal superantigens include the pyrogenic exotoxins A (and antigenic variants), C, G-J, SMEZ, and SSA. Superantigens are implicated in several diseases including toxic shock syndrome, scarlet fever and food poisoning; and their function appears primarily to debilitate the host sufficiently to permit the causation of disease. Structural studies over the last 10 years have provided a great deal of information regarding the complex interactions of these molecules with their receptors. This, combined with the wealth of new information from genomics initiatives, have shown that, despite their common molecular architecture, superantigens are able to crosslink MHC class II molecules and T-cell receptors by a variety of subtly different ways through the use of various structural regions within each toxin.     

Erroneous thyroid-stimulating hormone radioimmunoassay results due to interfering antibovine thyroid-stimulating hormone antibodies.

Apparent elevation of serum thyroid-stimulating hormone (TSH) levels was found in a case of a patient receiving adequate replacement therapy after thyroidectomy for a follicular carcinoma. This was an artifact due to interference with the radioimmunoassay by circulating antibovine TSH antibodies. The double-antibody technic showed spuriously elevated levels, and the single-antibody technic showed low levels of serum TSH by radioimmunoassay in the presence of antibodies.     

致Graves病的人促甲状腺激素受体多肽位点筛查

目的通过检测人促甲状腺激素受体(thyroid-stimu lating hormone receptor,TSHR)多肽片段与人白细胞抗原(hum an leukocyte antigen,HLA)分子的结合力,寻找人TSHR作为自身抗原导致G raves病的多肽位点。方法应用ELISA方法检测31条人工合成的膜外区人TSHR多肽片段与8种HLA分子的结合,通过计算IC50值,确定能与G raves病易感基因HLA-DR3和HLA-DQA1*0501高亲和力结合的多肽片段,并检测它们与G raves病保护基因HLA-DR7及HLA-DQA1*0201的结合能力。结果合成的31条TSHR多肽片段与HLA-DR3及-DR7的结合能力相近,且半数以上的多肽片段能与HLA-DQA1*0501及-DQA1*0201高亲和力结合。人TSHR多肽片段与HLA-DR分子结合能力为DR2>DR5>DR18>DR3/DR7>DR1,而与HLA-DQ分子的结合则为DQA1*0201>DQA1*0501。人TSHR多肽片段183~198、195~210、248~263、301~320、343~362及357~376能同时与HLA-DR3及HLA-DQA1*0501高亲和力结合,IC50值均<1μmol/L;而它们与HLA-DR7及HLA-DQA1*0201的结合力低。结论膜外区人TSHR多肽片段183~198、195~210、248~263、301~320、343~362及357~376有可能为导致G raves病的自身抗原多肽位点。     

Abnormal structure-function relationships in hereditary dystonia

Primary torsion dystonia (PTD) is a chronic movement disorder manifested clinically by focal or generalized sustained muscle contractions, postures, and/or involuntary movements. The most common inherited form of PTD is associated with the DYT1 mutation on chromosome 9q34. A less frequent form is linked to the DYT6 locus on chromosome 8q21-22. Both forms are autosomal dominant with incomplete (∼30%) clinical penetrance. Extensive functional and microstructural imaging with positron emission tomography (PET) and diffusion tensor MRI (DTI) has been performed on manifesting and non-manifesting carriers of these mutations. The results are consistent with the view of PTD as a neurodevelopmental circuit disorder involving cortico-striatal-pallido-thalamocortical (CSPTC) and related cerebellar–thalamo-cortical pathways. Studies of resting regional metabolism have revealed consistent abnormalities in PTD involving multiple interconnected elements of these circuits. In gene carriers, changes in specific subsets of these regions have been found to relate to genotype, phenotype, or both. For instance, genotypic abnormalities in striatal metabolic activity parallel previously reported reductions in local D₂ receptor availability. Likewise, we have identified a unique penetrance-related metabolic network characterized by increases in the pre–supplementary motor area (SMA) and parietal association areas, associated with relative reductions in the cerebellum, brainstem, and ventral thalamus. Interestingly, metabolic activity in the hypermetabolic areas has recently been found to be modified by the penetrance regulating D216H polymorphism. The DTI data raise the possibility that metabolic abnormalities in mutation carriers reflect adaptive responses to developmental abnormalities in the intrinsic connectivity of the motor pathways. Moreover, findings of increased motor activation responses in these subjects are compatible with the reductions in cortical inhibition that have been observed in this disorder. Future research will focus on clarifying the relationship of these changes to clinical penetrance in dystonia mutation carriers, and the reversibility of disease-related functional abnormalities by treatment.     

Secretion of growth hormone and thyroid-stimulating hormone in patients with dementia

We studied the growth hormone (GH) response to GH-releasing hormone (GHRH) and the thyroid-stimulating hormone (TSH) response to thyrotropin-releasing hormone (TRH) in four groups of patients with dementia and examined whether GH and TSH secretion is altered in patients with Alzheimer's disease. The four groups included those with Alzheimer's disease (n=28), parkinsonism with dementia (n=10), progressive supranuclear palsy with dementia (n=10), and dementia of vascular origin (n=28). The results showed no differences among the four groups in GH response to GHRH (12.2 ± 2, 10.7 ± 2, 8.9 ±1.1, and 9.9 ± 1.9 g/ml, respectively); there was no correlation between GH response to GHRH and sex, stage of the disease, or cerebral atrophy. The proportion of patients with exaggerated, normal, or lower GH response was similar in the four groups. There were also no differences among the groups in terms of TSH response to TRH (9.2 ±0.9, 11.1 ± 1, 11.1 ± 1, and 10.3 ± 1 mU/ml, respectively), nor was there a correlation between TSH response to TRH and sex, stage of the disease, cerebral atrophy, or GH response to GHRH. The proportion of those with exaggerated, normal, or lower TSH response was similar in the four groups. Cerebrospinal somatostatin levels were similar in Alzheimer's disease and vascular dementia patients. These findings indicate that neither GH response to GHRH nor TSH response to TRH provides a useful diagnostic adjunt in Alzheimer's disease patients.Abbreviations AD Alzheimer's disease - PD parkinsonism with dementia - PSP progressive supranuclear palsy - VD dementia of vascular origin - GH growth hormone - GHRH growth hormone releasing hormone - TRH thyrotropin releasing hormone - TSH thyroid stimulating hormone Correspondence to: J.M. Gomez     

Serum thyroid-stimulating hormone in cerebrovascular disease

A thyrotropin-releasing hormone (TRH) test with serum thyroid-stimulating hormone (TSH) assays was performed in 22 euthyroid stroke patients without thyroid disease and the results were compared with those in 17 age-matched euthyroid controls. Basal and maximum TSH levels after TRH injection were significantly lower in the stroke group without elevation of basal serum thyroid hormone levels. There was a tendency towards an inverse relationship between TSH levels and the degree of pareses of the extremities. The test was repeated in 7 stroke patients 3-4 months after the onset of stroke with essentially the same results. The abnormal TSH parameters in stroke patients seem to be the result of the brain lesion per se.     

Recent structure-function relationships in normal and injured mammalian kidneys

Recent investigations have been aimed at understanding the ultrastructural-functional relationships within the kidney at organ, tubule, subcellular, and molecular levels. This has led to a redefinition and more precise segmentation of the renal tubule. For example, the connecting piece between distal tubule and collecting system has now been established. The use of immunocytochemical techniques, such as fluorescence, ferritin- or peroxidase-labelled immunoglobulin methods, has made it possible to identify proteins in the kidney especially in renal corpuscles. Two major noncollagenous glycoproteins, fibronectin and laminin, have now been identified in the glomerulus. The glycosaminoglycan, heparan sulfate, has been localized to the glomerular basement membrane and is thought to play an important role in charge perm-selectivity during glomerular filtration. Subtle changes in glomerular podocyte or endothelial cell structure are postulated by some to play a role in the pathogenesis of acute renal failure. The role of the mesangial cell in glomerular function is being studied in situ in homogeneous cell populations. These cells are capable of prostaglandin production and can contract in response to hormonal stimulation. The intimate positioning of short- and long-looped nephrons in the renal medulla and the unique nature of the pelvic epithelium correlates well with the purported role of urea recycling in the urinary concentrating mechanism. Determination of elemental concentration of soluble substances in various renal cell and extracellular compartments have been made using freeze-hydrated and freeze-dried cryosections of kidney tissue. The medullary and cortical ascending thick limbs of the distal tubule are morphologically and functionally distinct regions. Their response to hormonal stimulation and their enzymatic activities are quite different. Morphological studies of the collecting duct have provided new insight into the role this segment of the uriniferous tubule plays in fluid and electrolyte transport and urinary acidification.     

Rapid detection of a point mutation in thyroid-stimulating hormone beta-subunit gene causing congenital isolated thyroid-stimulating hormone deficiency.

Previous study showed that congenital isolated TSH deficiency in Japan is resulted exclusively from a G-A transition at nucleotide 145 in exon 2 of the TSH beta-subunit gene. All reported cases were from the inbred in Shikoku Island. We describe here a 10-year-old boy with hereditary TSH deficiency in the same area. The patient was born with a weight of 3,225 g to non-consanguineous parents. Evaluation at age 2 months revealed typical manifestations of cretinism without goiter. Serum T4, T3, and TSH values were 2.53 micrograms/dl, 107 ng/dl, and 0.5 microU/ml, respectively. A TRH stimulation test showed no increment of serum TSH value. Other anterior pituitary hormone levels were all within the normal range. Two oligonucleotide primers T1a and T1b were synthesized according to the sequence data. Amplified 169 bp nucleotides in exon 2 of the TSH beta gene with this primer set were digested with MaeI. Both the phenotypically normal brother and normal controls showed only the 169 bp fragment, whereas the proband showed 140 and 29 bp fragments and both parents showed three fragments; 169, 140, and 29 bp. These results were consistent with the point mutation of TSH beta gene in Japanese patients with congenital isolated TSH deficiency. Our PCR method with MaeI digestion contributes to the rapid detection of the homozygous patient and the heterozygous carrier.     

Comparative connective tissue structure-function relationships in biologic pumps

A complex connective tissue framework exists in mammalian hearts that surrounds and interconnects individual myocytes and fascicles of cells. Recent evidence suggests that this connective tissue plays a role in maintaining shape, modulating contractile forces, and mediating elastic recoil during cavity filling and contraction. In order to analyze the involvement of connective tissue in pump contraction and recoil, we examined silver impregnated connective tissue in rat hearts which spontaneously jet through fluid ex vivo by contracting their cavities forcefully and then sucking fluid for the next cycle, and compared them to frog hearts which beat actively under the same conditions, but do not demonstrate jet propulsion. A further analysis was carried out in unrelated but analogous models: the squid and octopus. The former jets rapidly through the ocean, while the latter moves sinuously along the seabed. We observed highly interconnected myocytes in the rat heart, whereas frog myocytes are individually wrapped by connective tissue but are not interconnected. The squid mantle muscle is surrounded by a complex connective tissue grid that is tethered to each muscle cell, whereas the octopus mantle muscle cells are surrounded by connective tissue but are not tethered. These observations suggest that myocyte connective tissue tethering may be necessary for muscle cavities to generate forceful and coordinated contractions sufficient for rapid ejection and suction of fluid.     

Maturation of the secretion of thyroid hormone and thyroid-stimulating hormone in the fetus

BACKGROUND. Data on human fetal thyroid function have largely been derived from histologic studies or studies of cord-blood samples obtained at hysterotomy or delivery. These data may not represent true normal values. Cordocentesis (ultrasound-guided blood sampling from the umbilical cord) is a technique that allows investigation of physiologic processes in fetuses not under stress. METHODS. We measured serum thyroid-stimulating hormone, total and free thyroxine (T4), total and free triiodothyronine (T3), and thyroxine-binding globulin in blood samples from 62 fetuses. The samples were obtained by cordocentesis (n = 58) or cardiocentesis (n = 4) at 12 to 37 weeks of gestation. Maternal serum samples were obtained immediately before fetal blood sampling. RESULTS. Fetal serum thyroid-stimulating hormone, thyroxine-binding globulin, and total and free T4 and T3 concentrations increased significantly with the length of gestation (P less than 0.001). The only significant association among these variables, independent of the length of gestation, was between thyroid-stimulating hormone and free T4 (P less than 0.0001). Maternal serum concentrations of these variables did not change during gestation, and there was no significant relation between fetal and maternal values. Most fetal serum concentrations of thyroid-stimulating hormone were higher, whereas most serum total and free T3 concentrations were lower than the respective values for normal adults. The fetal serum total T4, free T4, and thyroxine-binding globulin values reached the level of the mean adult values at approximately 36 weeks of gestation. CONCLUSIONS. The increases in fetal serum concentrations of thyroid-stimulating hormone, thyroxine-binding globulin, and total and free T4 and T3 during gestation reflect increasing maturation of the pituitary, thyroid, and liver. The finding of increasing fetal serum concentrations of thyroid-stimulating hormone in the presence of increasing thyroid hormone concentrations suggests that the sensitivity of the fetal pituitary gland to negative feedback is limited or is counterbalanced by increasing stimulation by thyrotropin-releasing hormone from the hypothalamus.     

Suppression of maternal pituitary thyroid-stimulating hormone during pregnancy

Maternal serum thyroid-stimulating hormone (TSH) has been estimatedin 38 pregnant women with three new specific non-radioactivemonoclonal immunoassays which can measure very low concentrations.Values for the Pharmacia-LKB ‘DELFIA’ fluoroimmunoassayand for the Amersham ‘Amerlite’ luminescence immunoassaywere statistically identical over the range 0.15–1.5 uIU/mlWHO 2nd IRP 80/558, but thereafter the Amersham values wereslightly lower. The Abbott ‘IMx’ assay system gaveslightly higher results commensurate with the quoted highernon-pregnant normal values. The Amersham assay was preferredon practical grounds. There was no relationship of TSH levelsto HCG levels, or to the length of pregnancy. Four women hadapparently zero TSH levels by the Amersham and Pharmacia-LKBassays, although the Abbott assay could detect very low concentrations.Together with women whose TSH levels were below the normal non-pregnantrange for each assay, there were a total of eight women (21%)with TSH levels below ‘normal’. This suppressionof maternal pituitary TSH levels during pregnancy was consideredto be due to the central feedback inhibitory thyrotrophic activityof HCG, as a separate placental thyrotrophic hormone is believednot to exist. The enlargement of the maternal thyroid glandand the increased production of thyroid hormones during normalpregnancy is likewise to be attributed to HCG rather than toan increase in TSH production.     

Structural determinants for G-protein activation and specificity in the third intracellular loop of the thyroid-stimulating hormone receptor

The selectivity of G-protein recognition is determined by the intracellular loops (ICLs) of seven-transmembrane-spanning receptors. In a previous study, we have shown that the N-terminal and central portions of ICL2 from F525 to D530 participate in dual -/-protein activation by the thyroid-stimulating hormone receptor (TSHR). ICL3 is another major determinant for G-protein activation. Therefore, the aim of our study was to identify important amino acids within ICL3 of the TSHR to gain insight in more detail about its specific function for - and -protein activation and selectivity. Single-alanine substitutions of residues in the N-terminal, middle, and C-terminal region of ICL3 were generated. N-terminal residues Y605 and V608 and C-terminal positions K618, K621, and I622 were identified as selectively important for activation, whereas mutations in the center of ICL3 had no effect on TSHR signaling. Our findings provide evidence for an amino acid pattern in the N- and C-terminal part of ICL3, which is involved in -mediated signaling. Furthermore, molecular modeling of interaction of TSHR ICL2 and 3 with suggests three potential contact sites: TSHR C-terminal ICL3 with β5-6 loop of , TSHR ICL2 residues I523-R531 with β2-3 loop and N-terminal helix of , and TSHR ICL2/transmembrane helix (TMH) 3+ICL3/TMH6 with C-terminal tail of .Maren Claus and Susanne Neumann contributed equally to this work.     

Molecular economy of nature with two thyrotropins from different parts of the pituitary: pars tuberalis thyroid-stimulating hormone and pars distalis thyroid-stimulating hormone

Thyrotropin (TSH) is classically known to be regulated by negative feedback from thyroid hormones and stimulated by thyrotropin-releasing hormone (TRH) from the hypothalamus. At the end of the 1990s, studies showed that thyrotroph cells from the pars tuberalis (PT) did not have TRH receptors and their TSH regulation was independent from TRH stimulation. Instead, PT-thyrotroph cells were shown to have melatonin-1 (MT-1) receptors and melatonin secretion from the pineal gland stimulates TSH-β subunit formation in PT. Electron microscopy examinations also revealed some important differences between PT and pars distalis (PD) thyrotrophs. PT-TSH also have low bioactivity in the peripheral circulation. Studies showed that they have different glycosylations and PT-TSH forms macro-TSH complexes in the periphery and has a longer half-life. Photoperiodism affects LH levels in animals via decreased melatonin causing increased TSH-β subunit expression and induction of deiodinase-2 (DIO-2) in the brain. Mammals need a light stimulus carried into the suprachiasmatic nucleus (which is a circadian clock) and then transferred to the pineal gland to synthesize melatonin, but birds have deep brain receptors and they are stimulated directly by light stimuli to have increased PT-TSH, without the need for melatonin. Photoperiodic regulations via TSH and DIO 2/3 also have a role in appetite, seasonal immune regulation, food intake and nest-making behaviour in animals. Since humans have no clear seasonal breeding period, such studies as recent ‘’domestication locus’’ studies in poultry are interesting. PT-TSH that works like a neurotransmitter in the brain may become an important target for future studies about humans.