Effect of TSH and melatonin on thyroid activity in the rat.

Melatonin and TSH, injected separately, caused no change of the blood thyroxine level at 30 min after treatment. Simultaneous or subsequent administration of the two hormones induced an increase of the level. Thus, melatonin is capable of potentiating acute, thyroxine mobilizing effect of TSH.     

Non-neurogenic stimulation of adrenomedullary secretion during endotoxicosis in the one day old rat

In both newborn and twenty day old rats, bacterial (S. enteritidis) endotoxin caused a marked decrease in adrenal epinephrine and norepinephrine. Catecholamine release was prevented by ganglionic blockade in 20 day old, but not 1 day old animals, indicating that the release in the neonatal rat is "non-neurogenic." The amount released was physiologically significant: equivalent to 20 micrograms/kg by i.v. administration. Two possible mechanism known to promote non-neurogenic secretion are hypoxia or histamine release, both of which can occur during endotoxicosis.     

Effect of melatonin on steroid production by rat adrenals under in vitro superfusion conditions

In vitro superfusion of adrenals from male and female Wistar rats resulted in a gradual decline of the corticosterone content in the samples collected for a period of 80 minutes. ACTH administration either in the beginning or in the end of the perfusion period led to a marked increase in the level of this steroid. Contrary to this, progesterone content in the collected perfusates was constant throughout the experiments and was not influenced by ACTH. The quantities of testosterone secreted under these conditions were below the sensitivity of the assay. Perfusion with melatonin (200 pg/ml) did not affect corticosterone secretion, but resulted in a significant increase of the progesterone content in the recovered fractions. This effect was immediate and was followed by a decline in the progesterone concentration, observed during melatonin perfusion.     

Influence of VIP on thyroid hormone secretion

Since VIP occurs in intrathyroidal nerves its role in thyroid hormone secretion has been investigated. It has been found that VIP is a stimulator of iodothyronine secretion in mice. In this respect VIP has a weaker potency than TSH, but shows a similar time characteristic. Also, VIP and TSH potentiate each others effects. In contrast to the effect of TSH, that of VIP is uninfluenced by alpha-adrenoceptor blockade. VIP, like TSH, stimulates thyroid cyclic AMP production. Thus, VIP nerves might, together with TSH, adrenergic and cholinergic nerves and other peptides such as somatostatin, participate in the complex regulation of iodothyronine secretion. Beside this, VIP has also been found to stimulate calcitonin secretion in rats. Other intrathyroidal neuropeptides, such as substance P and CCK-4, have been found to be without effects on iodothyronine secretion, but, like VIP, to stimulate calcitonin secretion.     

Interaction between inhibin and melatonin on secretion of rat pituitary gonadotrophins

In presence of melatonin, the suppression of FSH levels by inhibin is reduced (31%) as compared to inhibin given alone (51%) to adult male rats. Likewise, inhibin decreases the prolactin enhancement following melatonin administration. These experiments demonstrate interaction of inhibin and melatonin on the secretion of pituitary gonadotrophins.     

Effect of melatonin on reproductive system in male rat, treated with estrogen in the first day of life.

The study aimed at demonstrating the possible stimulating or inhibitory effect of melatonin on reproductive system of male rats, the function of which was altered by single diethylstilbestrol dose introduced in the first day of life. The single estrogen dose in the first day of life induced in the studied rats, after they reached the adult age, inhibition of spermatogenesis as well as morphological and functional alterations in accessory sexual glands. This was associated with high level of LH gonadotrophin and with lowered testosterone level in the serum. Additional administration of melatonin between 45th and 84th day of life accentuated the above described changes induced by the single estrogen injection. Results of the studies demonstrated that melatonin in the experimental model not only failed to stimulate but provided an additional inhibitory effect on reproductive system in male rat.     

Influence of D-thyroxine on plasma thyroid hormone levels and TSH secretion.

Triiodothyronine (T3), thyroxine (T4), basal TSH and TSH after stimulation with TRH were determined in healthy subjects and patients treated with D-thyroxine (DT4). After a dosage of 6 mg DT4 the D/L T4 plasma concentration rose about 4-fold 4 hours after application and was only moderately elevated 14 hours later. To achieve constantly elevated T4 levels 3 mg DT4 were applied in the further experiment every 12 hours. The D/L T4 plasma concentration rose 2.5-4-fold and there was a small but significant increase of the D/L T3 plasma concentration. 74 hours after onset of treatment basal TSH was below detectable limits and the increase of TSH 30 min after injection of 200 mug TRH (TRH test) was only about 15% compared to zero time. The time course of TSH suppression was investigated after treatment with DT4 and LT4 (single dosage of 3 mg). TRH-tests were performed before, 10, 26, 50 and 74 hours after the first dosage of D or LT4. There was no difference in the time course of basal TSH and TSH stimulated by TRH. In 10 patients on DT4 long-term therapy, basal and stimulated TSH were found to be below the detectable limits of 0.4 mug/ml. Our results show that (1) plasma half-life of DT4 is less than 1 day, (2) TSH suppression after D and LT4 treatment is very similar, and (3) in patients on long-term DT4 treatment, TSH plasma concentration is below detectable limits even after stimulation with TRH.     

Influence of pimozide on a TRH induced TSH secretion in the rat during food deprivation.

Adult Wistar rats food deprived for 3 days had lower basal levels of TSH compared to normal fed animals. An increase of these lower levels to normal values was obtained following a prolonged (injections during 3 consecutive days) or acute treatment (single injection) with pimozide (1 mg/injection). Blood samples obtained after the last or an only injection of pimozide contained profound increased prolactin levels. Prolactin increase was more than 100-fold in fed and more than 30-fold in starved rats following prolonged pimozide treatment and more than 25-fold and 10-fold following a single injection of pimozide. An injection of 250 ng of TRH increased plasma concentrations of TSH in all groups, but this increase was more pronounced in fasted rats injected with pimozide during 3 consecutive days. It is concluded that fasting results in a dopaminergic inhibition of the sensitivity of the thyrotrophs to a TRH challenge.     

Direct action of melatonin on testosterone and cyclic GMP production using rat testis tissue in vitro.

When $\text{Escherichia}\text{coli}$ is growing in steady state with doubling times of 40 minutes or 20 hours, during shift up conditions, or under conditions of relaxed control of RNA synthesis, all rRNA cistrons are transcribed equally.     

Production and secretion of C-19 steroids by rat and guinea pig adrenals

The concentrations of C-19 steroids were measured in guinea pig and rat adrenals before and after castration as well as after stimulation with adrenocorticotropin hormone (ACTH). Characterization of adrenal C-19 steroids was also carried out by isolation with high-performance liquid chromatography and gas chromatography/mass spectrometry (GC/MS). From radioimmunoassay (RIA) data, androstenedione (4-DIONE) and 11 beta hydroxyandrostenedione (11 beta-DIONE) were the major C-19 steroids found in guinea pig adrenals, and castration induced a decrease of 4-DIONE levels only while all other C-19 steroids remained unchanged. In rat adrenals, the major C-19 steroids were 4-DIONE and testosterone, and they were also markedly inhibited after castration. With the exception of 11 beta-DIONE, all other C-19 steroids in circulation were eliminated after castration in both animals species. After ACTH administration in the guinea pig, adrenal 4-DIONE and 11 beta-DIONE levels were markedly stimulated, while an increase of only 11 beta-DIONE was observed in plasma. In the rat, ACTH had a small stimulatory effect on adrenal 52-androstane-3 alpha, 17 beta-diol (3 alpha-DIOL) and plasma 11 beta-DIONE levels. Analysis of guinea pig adrenal steroids by GC/MS confirmed the presence of C-19 steroids in adrenals (namely, 4-DIONE and 11 beta-DIONE) while, in the rat, this could not be confirmed. Our data indicate that production of C-19 steroids occurs in guinea pig adrenals, and 11 beta-DIONE is the major C-19 steroid as well as the only C-19 steroid secreted into the circulation. In the rat, the production of C-19 steroids detected by RIA is not supported by GC/MS data.     

Influence of exogenous melatonin and altered day length on reproductive performance of Polypay ewes

Two experiments were designed to evaluate postlight treatment use of melatonin as a method of overcoming photorefractory response by measuring the lambing percentage of early-postpartum Polypay ewes bred either late in the breeding season or during anestrus. In Trial 1, pregnant ewes (n = 140) were assigned to one of three groups: 1) ambient photoperiod (control), 2) extended light (20 h) from October 21 to December 27, and 3) extended light as in Group 2, followed by supplemental feeding of 10 mg melatonin/head (hd) daily until April 6. Breeding started on February 3 and ended April 9. A greater percentage of ewes given extended light plus melatonin (54%) and extended light alone (45%) lambed than controls (24%) (P = 0.06). In Trial 2, pregnant ewes (n = 158) were assigned to groups as in Trial 1, except extended light was given to Groups 2 and 3 from January 1 to March 11, and Group 3 ewes were supplemented with 10 mg melatonin daily from March 12 to June 10. Breeding started April 18 and ended June 10. Lambing percentage was increased (P < 0.01) by extended light plus melatonin (54%) compared to controls (6%) or ewes given only extended light (10%). The shift from artificially extended light to the shorter ambient light with or without melatonin enhanced the lambing percentage of early postpartum ewes on an accelerated lambing program during breeding late in the season. However, only the more pronounced shift from long days to short days, accomplished with extended light plus melatonin, was effective in inducing out-of-season breeding in Polypay ewes.     

Influence of thyroid status on Ca2+ mobilization and amylase secretion in rat pancreatic acini

Thyroid hormones influence Ca2+ homeostasis in both skeletal and cardiac muscle. Since secretory cells, like muscle cells, store and use Ca2+ in stimulus-response coupling, we have studied the effects of thyroid status on Ca2+ mobilization and secretion in a model secretory tissue, the pancreatic acinar cell. Hyperthyroidism was induced by rats by daily, subcutaneous injections of triiodothyronine for 8 days and hypothyroidism by adding 6-n-propyl-2-thiouracil to the drinking water for 14 days. Pancreatic acini were prepared by collagenase digestion of pancreatic tissue from hyper- and hypo-thyroid animals and from euthyroid controls. Ca2(+)-mobilization was assessed using Quin-2 fluorescence and secretion by assaying amylase release. The data indicate that the amount of Ca2+ mobilized by the muscarinic agonist carbachol or by cholecystokinin octapeptide increases with increasing thyroid hormone concentrations. Only in hypothyroidism was this change in Ca2+ homeostasis reflected by a parallel change in amylase secretion. This implies the existence of some compensatory mechanism which stabilizes secretory rate in the face of stimulus-evoked increases in intracellular Ca2+ concentration.     

Heparin-releasable lipase activity of rat adrenals, ovaries and testes.

The presence of NaCl-resistant, neutral triacylglycerol hydrolase (lipase) activity in rat adrenal gland, ovary and testis was studied. Both adrenals and ovaries but not testes were found to contain such a lipase. The activity of the enzyme in the adrenal gland was lowered during cortisol treatment and hypothyroidism. An elevated adrenal lipase activity was found during hyperthyroidism. Pseudo-pregnant and lactating rats had higher ovarian lipase activities than cyclic rats. Ovarian lipase activity in lactating rats was positively correlated with the serum concentrations of progesterone and of 20 alpha-hydroxyprogesterone and negatively correlated with the high-density-lipoprotein non-esterified cholesterol concentration. The lipase activity of adrenals and of ovaries was largely releasable from these organs by heparin and could be inhibited by an antibody against heparin-releasable liver lipase. This indicated that the lipase is extracellularly located and is similar to 'liver' lipase. A possible role of this lipase in adrenals and ovaries is discussed.