Correlation between 3′,5′, c-AMP levels and thyrotropin in separated rat pituitary thyrotropic cells

Summary The correlation between 3,5, c-AMP levels, TSH content and secretion of separated thyrotropic cells was studied. Incubation of the separated cells with 1, 10 and 100 ng of TRH does not change the 3,5, c-AMP levels, despite the significant rises of the TSH level. Dibutyryl c-AMP causes rise in TSH content, with no indication of its secretion. PGE₂ 10^–5 increased 3, 5, c-AMP levels with no change in the content or secretion of TSH in separated thyrotropic cells.     

Immunohistochemical individualization of adenohypophyseal thyrotropic cells in the monkey Macaca irus, using a human anti-beta-TSH antibody

Using the indirect immunofluorescence technique and an antiserum against the specific beta subunit of human TSH, selective immunocytochemical staining was localized in a cell population of the pars distalis of the monkey Macaca irus. These thyrotropic cells, which were positive to aldehyde fuchsin, Alcian blue, and PAS, were localized on the one hand in the ventro-medial zone of the pars distalis, especially alongside the large vessels of this region, and on the other in the zona tuberalis, i. e. at the bottom of the pars tuberalis. Some cells, sparsely distributed, were seen in the posterior part of the lateral lobes of the pars distalis, in the vicinity of the pars intermedia. No thyrotropic cells were encountered in pars tuberalis. Pars intermedia and posterior lobe contained no cells immunoreactive to beta-hTSH. In females, the thyrotropic cells were more numerous and larger than in males. Generally the thyrotropic cells were round or oval in shape, sometimes they were polygonal.     

Interaction between dehydroepiandrosterone, glucose-6-phosphate dehydrogenase, and cyclic adenosine-3',5'-monophosphate in neoplastic and normal human mammary tissue.

When total DHEA, G-6-PDH activity, and c-AMP were determined in human neoplastic mammary tissue and corresponding normal tissue the G-6-PDH activity in the former tissue greatly exceeded that found in normal tissue. On the other hand, a remarkable decrease of total DHEA and c-AMP could be detected in cancer tissue, hinting at the participation of DHEA in the intracellular regulation of G-6-PDH and c-AMP levels.     

Respective roles of circulating T4 and T3 in control of TSH secretion in severely iodide-deficient rats

Summary After a 6-month iodide deficiency, Wistar male rats were submitted to a normal iodine diet (20 and 50 g of¹²⁷I daily). Plasma T₃, T₄ and TSH were determined by RIA from 0 to 140 days of iodide refeeding. A highly significant correlation was found between plasma TSH and T₄ concentrations, but not between plasma TSH and T₃ levels. These data suggest that an increase in plasma T₃ alone, up to the normal value, is not able to inhibit TSH secretion. It is only when a certain plasma T₄ concentration is also reached, resulting in further T₃ formation through deiodination, that TSH secretion is inhibited.Dedicated to the memory of Prof. C. Simon.This work was supported by grants from the CNRS (Equipe de Recherche Associée no 234) and from the INSERM (A.T.P. 49.77.8).Acknowledgments. The authors wish to thank Mrs M. Chartier for valuable technical assistance. They are indebted to the rat pituitary distribution programme of NIAMDD, NIH, Bethesda, for their gift of rat TSH reagents.     

Antiproliferative effects of 4',5'-unsaturated adenine nucleosides on leukemia L1210 cells in vitro

Several 4',5'-unsaturated adenine nucleosides were shown to have antiproliferative activity against L1210 leukemia cells in vitro. The active nucleosides were cytotoxic to the L1210 cells as demonstrated by Trypan Blue uptake. The cytotoxicity was not induced by alterations in the ribonucleoside and deoxyribonucleoside triphosphate levels of the L1210 cells.     

Free and sulfoconjugated dehydroepiandrosterone, cyclic adenosime-3',5'-monophosphate, and free estriol in maternal and cord blood.

When free DHEA, its sulfatide, and sulfate were assayed in maternal plasma as well as in umbilical cord arterial and venous plasma, rather high concentrations were found in either fraction from cord arterial plasma, reflecting the fetal contribution not only of free DHEA and DHEA sulfate, but also of the lipophile steroid sulfatide. Since high DHEA levels were associated with elevated c-AMP concentrations, a certain interrelationship of both parameters is indicated. In the course of delivery, a rapid decrease of free estriol in maternal plasma was observed. Higher concentration of free estriol in umbilical venous plasma pointed at its placental biosynthesis from fetal precursors.     

Renal effects of 8-substituted derivatives of adenosine 3',5'-cyclic monophosphate in dogs

The renal effects of nine, 8-substituted derivatives of 3',5'-cyclic monophosphate were studied in anesthetized mongrel dogs. Infusion of the compounds into a renal artery resulted in an increase of renal blood flow without any effect on blood pressure. Diuretic and natriuretic effects are evident with 6 of the 9 derivatives. As these 8-substituted analogues exert renal effects in a manner similar to that seen with the parent nucleotide, cyclic adenosine 3',5'-monophosphate, they may serve as useful pharmacological agents in vivo.     

IgG purification to measure the level of an iodinated thyroglobulin peptide, the 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine in human serum

Antibodies reacting with 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine (I2Tyr-I2Tyr) were elicited in rabbits by immunization with an oxidized yeast conjugate coupled with I2Tyr-I2Tyr. Ion-exchange chromatography was used to purify immunoglobulins, in order to improve the specificity in measurement of I2Tyr-I2Tyr level in patient serum. IgG binding capacity versus I2Tyr-I2Tyr was considerably increased after immunoglobulin purification.     

Control of pituitary secretion of melanotropin in an anuran amphibian by thyrotropin releasing factor (TRH). Study in vitro]

Intermediate lobes from Rana esculenta pituitary glands were continuously superfused for 7 hrs at 28 degrees C with amphibian culture medium. alpha-MSH release was measured by use of a sensitive double antibody radio-immunoassay system. alpha-MSH secretion was inhibited by low temperatures. A large increase in alpha-MSH release was observed when Thyrotropin Releasing Hormone (TRH) at doses ranging from 10(-9) to 10(-7) molar was added to the superfusion medium. Since large amounts of TRH are to be found in the hypothalamus of amphibians but have no effect over pituitary TSH secretion, the action of TRH over alpha-MSH release may have a physiological significance.     

Inhibition of TSH-stimulated thyroid hormone release and potentiation of TRH-stimulated TSH release by indomethacin in perifusion systems of rat thyroids and pituitaries

Using indomethacin (Ind), a prostaglandin synthesis inhibitor, in vivo experiments in rats and in vitro experiments with perifusion systems of rat thyroids and pituitaries were conducted. After 35 days of intragastric infusion of Ind, serum TSH levels were markedly increased, the thyroid was swollen and, as a consequence, T3 and T4 levels were normal. The T3 release from perifused rat thyroids under continuous stimulation with 10 mU/ml TSH was inhibited significantly (p less than 0.01) by 1.0 X 10(-6) M Ind. On the other hand, the TSH release from perifused rat pituitaries under TRH stimulation was enhanced conspicuously by Ind. It was concluded that Ind decelerated thyroid hormone release from the thyroid and accelerated TSH release from the pituitary in perifusion systems.     

Pancreastatin,a chromogranin A-derived peptide,inhibits leptin and enhances UCP-2 expression in isolated rat adipocytes

Leptin, the ob gene product, is an adipocyte-secreted hormone that centrally regulates weight by decreasing caloric intake and increasing energy expenditure. Expression of leptin is regulated by dietary status, insulin, glucocorticoids and catecholamines. Pancreastatin (PST), a chromogranin A-derived peptide, correlates with catecholamine levels, and may play a role in the physiology of stress, modulating endocrine secretion and metabolism. Thus, PST has been found to exert a lipolytic and anti-insulin effect in white adipocytes. The aim of the present work was to investigate a possible role of PST modulating the expression of key genes involved in lipid storage and metabolism: leptin, PPAR-2, UCP-1 and UCP-2. We incubated isolated rat epididymal adipocytes with 100 nM PST for 16 and 24 h. Leptin, UCP-2 and UCP-1 mRNA levels were assessed by RT-PCR, followed by Southern blot. Leptin secretion was also measured by ELISA. PST inhibited leptin expression and secretion at 16-h incubation, but this effect was no longer observed after 24 h. On the other hand, PST stimulated the expression of UCP-2 after 16 h. However, the effect was still significant after 24 h. The inhibitory effect of PST on leptin expression and secretion and the stimulation of UCP-2 expression were prevented by blocking PKC. UCP-1 and PPR-2 expression did not change after PST stimulation. Leptin differentially regulates the expression of key genes in the rat adipocyte, upregulating the expression of UCP-2 and inhibiting the expression and secretion of leptin by a mechanism that involves PKC activity. These effects may contribute to the metabolic action of catecholamines in physiological and pathophysiological conditions with increased sympathetic activity.Received 5 September 2003; received after revision 6 October 2003; accepted 14 October 2003     

Inhibition of TSH-stimulated thyroid hormone release and potentiation of TRH-stimulated TSH release by indomethacin in perifusion systems of rat thyroids and pituitaries

Summary Using indomethacin (Ind), a prostaglandin, synthesis inhibitor, in vivo experiments in rats and in vitro experiments with perifusion systems of rat thyroids and pituitaries were conducted. After 35 days of intragastric infusion of Ind, serum TSH levels were markedly increased, the thyroid was swollen and, as a consequence, T₃ and T₄ levels were normal. The T₃ release from perifused rat thyroids under continuous stimulation with 10 mU/ml TSH was inhibited significantly (p<0.01) by 1.0×10^–6 M Ind. On the other hand, the TSH release from perifused rat pituitaries under TRH stimulation was enhanced conspicuously by Ind. It was concluded that Ind decelerated thyroid hormone release from the thyroid and accelerated TSH release from the pituitary in perifusion systems.     

Inhibition of the 3' to 5' exonuclease activity of the DNA polymerase I of Escherichia coli by deoxyribonucleotides

The 3' à 5' exonuclease activity of E. coli DNA-polymerase I is inhibited by nucleotides and deoxynucleotides at concentrations (< 1 mM) where polymerase activity is not affected. This inhibitory effect depends on the nature of the excised deoxynucleotide, excision of purines being much less inhibited than that of pyrimidines. It does not depend on the purine or pyrimidine nature of the inhibitor.     

A histochemical demonstration of the Na+ + K+-ATPase activity in the thyroid and the effect of cyclic adenosine monophosphate (c-AMP).

With a suitable modification of the Farquhar and Palade technique the Na+ + K+-ATPase activity in guinea-pig thyroid is demonstrated. The addition of c-AMP (5 X 10(-6) M or 1.5 X 10(-5) M) to the incubation media produced an apparent intensification of the Na+ + K+ -ATPase activity in the thyroid.     

Correlation of immunogenicity with suppression of lymphocyte adenosine 3',5'-monophosphate-dependent protein kinase.

Cyclic-AMP-dependent protein kinase activity was depressed in whole spleen as well as in isolated splenic lymphocytes from 3-methylcholanthrene (MCA), R3230 AdCa mammary adenocarcinoma, N-hydroxy-2-acetylaminofluorene, and 4-dimethylaminoazobenzene (DMAAB) tumor-bearing Fischer rats as compared to control animals. The magnitude of depression increased with the immunogenicity of the tumor. The depressed enzyme activity was the result of a reduced Vmax for adenosine 3',5'-monophosphate (cAMP)-stimulated histone phosphorylation.     

On the time course of thyrotropin suppression by high doses of thyroid hormones

Summary Basal and stimulated TSH decreased progressively. Basal TSH was suppressed below the detection limit of 0.4 U/ml after 74 h in 2 of the T₃ and all of the T₄ treated indiduals. At this time, in both groups 3 individuals could be significantly stimulated by TRH (about 5% of the pretreatment stimulation). There was no significant difference in the time course of suppression obtained by T₃ or T₄, though plasma T₃ levels in the T₄ treated group were considerably lower.Supported by the Deutsche ForschungsgemeinschaftThe excellent technical assistance of MissM. Knöpfle, MissU. Neun and Mr.G. Magin is gratefully acknowledged.     

Reciprocal changes in serum free thyroxine fraction and thyrotropin level after administration of thiocyanate to rats

Summary The administration of thiocyanate to rats caused a significant increase of serum free thyroxine fraction, which coincided with the significant decrease of TSH level. The other components (AFT₄, T₄, T₃) in serum at this time were decreased or unchanged. The finding suggests the role of free thyroxine fraction in feed-back regulation of TSH secretion.Acknowledgment. We thank Dr A. Parlow and the NIAMDD, Rat Pituitary Hormone Distribution Program, for material for rat TSH immunoassay; Dr J. Nauman (Inst. Postgrad. Med., Warsaw, Poland) for T₃ antibody, and to Ing. J. Sadlo, Mrs M. t'astná and Miss R Fajkoová for technical assistance.     

Melatonin biosynthesis in the mammalian pineal gland

Summary Rhythmic production of melatonin by the mammalian pineal occurs in response to noradrenergic stimulation which produces a cascade of biochemical events within the pinealocyte. In the rat, massive changes in NAT activity result from an increase in intracellular c-AMP levels produced by a synergistic interaction whereby an ₁ activation amplifies -adrenergic stimulation. The intracellular events mediating this effect are described. A major aspect of the temporal control of melatonin production is the programmed down-regulation of responses to noradrenergic stimulation once the initial surge of c-AMP is produced. Noradrenergic activation of the gland also influences other enzymic functions, including tryptophan hydroxylase and HIOMT activities, and produces a dramatic increase in intracellular c-GMP levels. Other neurotransmitters and neuropeptides, e.g. VIP, may also influence pineal function and comparisons are, made between the rat, the subject of the bulk of experimental studies, and other species.     

An immunofluorescent method for identification of isolated thyrotropic cells.

A sensitive and specific immunofluorescent method for identification of thyrotropic cells was developed. The TSH-producing cells were found to be heterogenic in their morphology and intensity of staining.