Search Results

Abstract

The aims of the present study were to determine the influence of brief subclinical hypothyroidism on the isoforms of serum thyrotropin (TSH) and to examine the net charge of TSH in different metabolic states. Sera were obtained from euthyroid subjects (n=7) and from patients with subclinical hypothyroidism (n=8) before and 30 min after the intravenous administration of 200 μg thyrotropin-releasing hormone (TRH). The TSH from human pituitary extracts (IRP 68/38), basal and TRH-stimulated serum TSH was immunoconcentrated and further analysed by isoelectric focusing (IEF) and lentil lectin affinity chromatography. TSH immunoreactivity was determined in each specimen or fraction with an automated highly sensitive chemiluminometric TSH assay. We found that basal TSH in subclinical hypothyroidism, and TRH-released TSH in euthyroidism and in subclinical hypothyroidism is distributed in a similar neutral to acidic pattern, which significantly differs from the more alkaline to neutral isoform pattern of intrapituitary TSH (P<0·05). IEF analysis of pituitary standard TSH revealed 3 major peaks (pI values 7·5; 6·6; 5·8) whereas in most euthyroid or subclinically hypothyroid subjects 5 peaks were found. Lentil lectin affinity chromatography revealed that TRH-released TSH in euthyroid subjects has more core fucose residues than TSH from patients with subclinical hypothyroidism (64·6 ±6·7 vs 12·5 ±2·7%, P<0·0001).

Thus pituitary standard TSH seems to be less mature material than circulating TSH. Perhaps no alteration in the IEF pattern of TSH was detected during early hypothyroidism because sialylation of TSH was increasing as sulfatation was decreasing. Nevertheless, a change in the core fucose content of TSH was detectable by lentil analysis.

Journal of Endocrinology (1995) 144, 561–567

ABSTRACT

A screen of a range of bacteria normally found in gut flora identified eight with the ability to bind TSH specifically. These included the previously reported Yersinia enterocolitica, Gram-positive, Gramnegative, pathogenic and commensal organisms. Eleven preparations of TSH-receptor autoantibodies strongly able to displace ¹²⁵I-labelled TSH from the mammalian TSH receptor differed in their ability to displace the tracer from binding to bacterial extracts. None could displace the tracer from E. coli 06–1, four displaced ¹²⁵I-labelled TSH from E. coli V21/1 and five displaced the tracer from Y. enterocolitica. Of those immunoglobulin preparations which did react with the bacterial protein, their apparent potency compared with that of TSH in displacing tracer from bacterial binders was an order of magnitude greater than with the mammalian receptor. This is consistent with the autoantibodies having a relatively better fit with the bacterial antigen than with the receptor when compared with TSH. The bacterial-binding activity and mammalian receptor-binding activities in each of two samples co-chromatographed on a Remazol yellow GGL–Sepharose affinity column strongly indicated that the same immunoglobulin species reacts with both antigens. These results are consistent with the proposal that a bacterial protein is the primary immunogen for the TSH-receptor antibodies in at least some patients with Graves' disease.

Journal of Endocrinology (1989) 121, 571–577

ABSTRACT

Mitotic and labelling indices were studied in the thyroid follicular cells of the male rat within the first hour after a single injection of TSH or TSH vehicle, using the metaphase arrest agent vincristine sulphate. There was a significant increase in metaphase index over control values 5, 15, 30 and 60 min after injection of TSH. There was no significant change in the tritiated thymidine labelling index in TSH-treated rats in comparison with vehicle-injected controls. None of the metaphase figures was labelled, showing that G2 in this tissue is longer than 2 h.

Journal of Endocrinology (1989) 121, 27–30

Abstract

Inositol 1,4,5-trisphosphate (InsP₃) 3-kinase phosphorylates the Ca²⁺-mobilizing second messenger InsP₃ to inositol 1,3,4,5-tetrakisphosphate (InsP₄). InsP₃ 5-phosphatase dephosphorylates InsP₃ to inositol 1,4-bisphosphate (InsP₂). We compared the effects of TSH added to a culture of FRTL-5 thyroid cells on the activity of InsP₃ 5-phosphatase and InsP₃ 3-kinase. InsP₃ 3-kinase activity was decreased at a physiological concentration of TSH. Inhibition of this activity started after 3 h of incubation with TSH and was maximal after 24 h. In contrast, InsP₃ 5-phosphatase activity was not affected by TSH under the same conditions. The inhibitory effect of TSH on InsP₃ 3-kinase was characterized as follows: a) inhibition of activity was mimicked by both dibutyryl cyclic AMP and forskolin; b) activity obtained by mixing lysates of TSH-stimulated and non-stimulated cells was the sum of each activity measured separately; c) inhibition persisted after a crude lysate of TSH-stimulated cells had been subjected to SDS/polyacrylamide gel electrophoresis and the extraction of InsP₃ 3-kinase activity. The data suggest that TSH reduced the activity of InsP₃ 3-kinase in FRTL-5 cells either by a phosphorylation/dephosphorylation mechanism, or by affecting expression of the enzyme.

Journal of Endocrinology (1995) 144, 527–532