The effects of prolonged, intracerebroventricular prolactin treatment on luteinizing hormone secretion, catecholaminergic activity and estrous behavior in ewes.

Searching for the role of prolactin (PRL) in controlling gonadotropic axis activity in sheep, we studied the effects of prolonged, intracerebroventricular (i.c.v.) PRL infusion on luteinizing hormone (LH) secretion and catecholaminergic activity in the hypothalamic infundibular nuclei/median eminence (IN/ME) in sexually active ewes during the periovulatory period. Three groups of animals received the following treatments: 1). i.c.v. infusion of PRL at a dose of 200 microg/day (Lower dose, n = 5); 2). i.c.v. infusion of PRL at a dose of 400 microg/day (Higher dose, n = 6), and 3). i.c.v. infusion of the vehicle (control, n = 5). Each dose of PRL was infused in a pulsatile manner, 4 x 50 microg/h and 4 x 100 microg/h, in 30-min intervals, respectively, during four consecutive days before oncoming ovulation. The estrous behavior of ewes following treatments was also monitored as a determinant of the GnRH/LH surge. Two series of blood collections were made in every ewe, the first on the day preceding the infusion (day 0 of the experiment), the second on the day after the infusion (day 5 of the experiment). In addition, on day 5 of the experiment, perfusions of the IN/ME were made by the push-pull method, either in control or lower dose-treated animals. It was shown that a significant (p < 0.01, p < 0.001) increase in tonic LH secretion during the periovulatory period remained in ewes irrespective of the kind of infusion. No statistical differences were found in LH pulse frequency, amplitude, or in the length of the pulse when compared with values from day 0 and 5 of the experiment within each group. A significant (p < 0.001) increase in IN/ME perfusate concentrations of dopamine and noradrenaline metabolites was noted in PRL-treated ewes in comparison with those in the control. The estrous behavior in PRL-treated animals was delayed for a few days, 3.80 +/- 0.80 days at the lower dose (p < 0.01), and 2.83 +/- 0.98 days at the higher dose (p < 0.05) in comparison with the control, 0.20 +/- 0.20 days. These data indicate that maintenance of an increased PRL concentration within the central nervous system (CNS) for a few days before oncoming ovulation has no inhibitory effect on tonic LH secretion. A few-day shift of the preovulatory GnRH/LH surge, as determined by estrous behavior, might, however, be a consequence of the PRL-induced increase in catecholamine turnover in the IN/ME.

Short-term modulation of prolactin secretion by melatonin in anestrous ewes following dopamine- and opiate receptor blockade.

This study tested the hypothesis that the dopaminergic and opioidergic systems are not involved in the short-term stimulatory action of melatonin (MLT) on the secretion of prolactin in anestrous ewes. Thus, MLT should stimulate prolactin release after blockade of either dopamine (DA) or opiate receptors with specific antagonists at the level of the pituitary gland and central nervous system (CNS), respectively. During afternoon intracerebroventricular (icv.) infusion of MLT, the mean plasma prolactin concentration increased significantly (P < 0.001) as compared with the concentrations noted before and during the infusion of the vehicle (veh.). As a result of subcutaneous (s.c.) injection of sulpiride (SULP, DA antagonist), an increase in plasma prolactin concentration was observed, followed by a gradual decrease during the icv. infusion of the vehicle. MLT infused icv. significantly increased (P < 0.001) the secretion of prolactin in SULP + MLT-treated ewes, as compared with the concentration of prolactin noted during infusion of the vehicle in SULP + veh.-treated ewes. Naloxone (NAL, opioid antagonist) infused icv. did not significantly affect the secretion of prolactin, however, a significant (P < 0.01) increase in the concentration was observed after the infusion. In MLT + NAL-treated ewes, the plasma prolactin concentration increased significantly (P < 0.001) during the infusion, as compared with the concentration noted before and that in NAL-alone infused ewes. These results demonstrate that melatonin stimulates prolactin release after the pharmacological exclusion of the dopaminergic input with the DA antagonist sulpiride and also despite the presence of DA activity in the hypothalamus after NAL treatment. Secondly, endogenous opioid peptides are not a major component of this melatonin action.

Daily GnRH and LH secretion in ewes is not modified by exogenous melatonin during seasonal anestrus.

The effect of central, short-term melatonin administration on daily GnRH and LH secretion was studied in ewes during seasonal anestrus. Melatonin, in a total dose of 32 micrograms and the vehicle were perfused for 4 hours into the mediobasal hypothalamus/median eminence (MBH/ME). The mean GnRH concentration during perfusion with melatonin decreased significantly (P < 0.05), as compared to the concentration during the preceding perfusion with the vehicle only. This change resulted from high variations in GnRH concentration noted during the initial phase of perfusion rather than from an action of melatonin. Melatonin perfused into the MBH/ME did not significantly affect LH secretion. A higher dose of melatonin and vehicle were also infused intracerebroventricularly (icv.) in either intact (300 micrograms for 3 hours) or ovariectomized (OVX) ewes (400 micrograms for 4 hours, 100 micrograms/100 microliters/h). In the intact animals, melatonin did not significantly affect LH secretion. Interestingly, melatonin significantly decreased (P < 0.05) the number of LH peaks in OVX ewes. These results demonstrate that melatonin delivered for a few hours directly into the central nervous system did not affect either daily hypothalamic GnRH release or pituitary LH secretion in intact ewes during seasonal anestrus, but did modify pulsatile LH secretion in ewes deprived of the negative feedback of estradiol.

Melatonin modulation of the daily prolactin secretion in intact and ovariectomized ewes. Relation to a phase of the estrous cycle and to the presence of estradiol.

The aim of this study was to investigate whether melatonin might modulate the daily prolactin secretion in the ewe during a period of ovarian activity and, if so, whether this modulatory action of melatonin was related to the presence of estradiol in the organism. Ewes in the late follicular and luteal phase, as well as overiectomized ewes without (OVX) and after 7 days of estradiol injections (OVX+E2) were examined. Melatonin was infused into the third brain ventricle (100 microgram/100 microliter/h) from 14.00 to 18.00 h. The concentration of prolactin increased significantly during the infusion of melatonin in late follicular-phase ewes, but not in luteal-phase ewes, as compared to the concentration before the infusion: range from 204.0 +/- 31.7 to 272.2 +/- 50.1 ng/ml vs. range from 68.2 +/- 31.8 to 94.7 +/- 33.1 ng/ml (mean +/- SEM, n = 4, p < 0.01) and to the concentration noted during control infusions: range from 130.0 +/- 58.0 to 179.3 +/- 55.6 ng/ml (mean +/- SEM, n = 4, p < 0.05). In ovariectomized ewes, the concentration of prolactin during infusion of melatonin increased significantly, unrelated to the presence of estradiol, as compared to the concentration before infusion: range from 136.7 +/- 20.3 to 260.0 +/- 11.6 ng/ml vs. range from 41.6 +/- 2.6 to 152.3 +/- 14.6 ng/ml in OVX ewes (mean +/- SEM, n = 4, p < 0.01) and range from 161.5 +/- 66.5 to 250.2 +/- 24.3 ng/ml vs. range from 61.2 +/- 1.7 to 159.2 +/- 43.3 ng/ml in OVX+E2 ewes (mean +/- SEM, n = 4, p < 0.01). Concentrations during infusion of melatonin in OVX and OVX+E2 ewes were also significantly higher than during the control infusions: range from 7.2 +/- 1.7 to 22.2 +/- 4.1 ng/ml (mean +/- SEM, n = 4, p < 0.001). These results indicate that melatonin may affect the daily secretion of prolactin in ewes during the breeding season, and suggest that the variable response of prolactin to the melatonin signal in intact and ovariectomized ewes relates to the interaction between both ovarian steroids - estradiol and progesterone - and the prolactin-releasing factor.

Urinary excretion of oestrone sulphate and cortisol in early pregnant gilts treated with glucocorticoids.

The aim of this study was to determine rates of urinary excretion of oestrone sulphate and cortisol in early pregnant gilts that were untreated or treated with either dexamethasone, corn oil or hydrocortisone. Twenty Polish Landrace gilts were used. They were grouped immediately after mating as follows: Experiment I--Group 1 (5 gilts), control animals and Group 2 (5 gilts), injected i.m. with dexamethasone (30 micrograms/kg) at 12-h intervals from day 13 to day 22 of pregnancy; Experiment II--Group 3 (5 gilts), injected i.m. with corn oil at 12-h intervals from day 13 to day 22 of pregnancy and Group 4 (5 gilts), injected i.m. with hydrocortisone acetate (250 mg) at 12-h intervals from day 11 to day 20 of pregnancy. Gilts were placed in metabolic cages, and 24-h urine aliquots were collected from day 6 to day 32 of pregnancy. On days 34-36 of pregnancy gilts were slaughtered and clinical data were collected. Rates of urinary excretion of oestrone sulphate and cortisol were determined by enhanced chemiluminescence immunoassays. The urinary excretion of oestrone sulphate expressed in nmol/24 h and mumol/mol creatinine were significantly correlated. There was no correlation between cortisol expressed in nmol/24 h and mumol/mol creatinine (p > 0.5). A first significant increase of urinary oestrone sulphate excretion, expressed in nmol/24 h, on days 13-14 and a second one on days 19-20 of gestation occurred in control untreated and oil-treated gilts. The urinary excretion of oestrone sulphate reached maximum values between days 25 and 32 of gestation. In dexamethasone-treated gilts cortisol excretion significantly decreased on day 16, i.e. 3 days after injections of dexamethasone had commenced. The treatment with hydrocortisone resulted in a significantly increased cortisol excretion after the last injection of hydrocortisone. There were no relations between levels of urinary oestrone sulphate excretion expressed in nmol/24 h and the number of foetuses. When the urinary excretion of oestrone sulphate was expressed in mol/mol creatinine we found a positive relation between concentrations on day 20 of pregnancy and the number of foetuses. In one untreated gilt with a relatively high urinary excretion of cortisol (more than 200 nmol/24 h) a lower number of foetuses was found at autopsy. In conclusion, both dexamethasone and hydrocortisone treatment seemed to delay the first observed peak in oestrone sulphate in gilts without affecting the embryonic survival and the number of viable foetuses.

Natural and melatonin-stimulated changes in the circadian rhythm of prolactin secretion in the ewe during seasonal anestrus.

The relations between the circadian rhythms of melatonin and prolactin, and the effect of melatonin infused into the third ventricle or the mediobasal hypothalamus (MBH) on prolactin secretion and dopamine (DA) release were studied in anestrous ewes under increasing daylength conditions. The decreased amplitude and duration of nocturnal melatonin secretion were accompanied with changes in the daily pattern of prolactin secretion. Marked peaks of prolactin occurred after sunrise and melatonin decreases in February, as well as significant evening or nocturnal peaks under long day conditions between April and July. Melatonin infused into the third ventricle evoked an abrupt increase in the concentration of prolactin after 30 min, and the enhanced prolactin level was significantly higher than during the control infusion (range from 204 +/- 75 to 248 +/- 48 ng/ml vs. 128 +/- 68 to 149 +/- 93 ng/ml, mean +/- SD). The concentrations of DA in MBH perfusates decreased during perfusion of melatonin but to a degree similar to that noted in untreated ewes. These data suggest that short-term infusions of melatonin stimulate the secretion of prolactin in the ewe under increasing daylength conditions, and that this effect is not mediated by changes in DA release.

Effect of glucocorticoid treatment on biochemical and hormonal blood parameters in early pregnant gilts.

Twenty Polish Landrace gilts were grouped immediately after mating as follows: Experiment I-- Group 1 (5 gilts), control animals and Group 2 (5 gilts), injected i.m. with dexamethasone (30 mg/kg) at 12-h intervals from day 13 to day 22 of pregnancy; Experiment II--Group 3 (5 gilts), injected i.m. with corn oil from day 13 to day 22 of pregnancy and Group 4 (5 gilts), injected i.m. with hydrocortisone acetate (250 mg) at 12-h intervals from day 11 to day 20 of pregnancy. Gilts were placed in metabolic cages on day 7. On days 34-36 of pregnancy gilts were slaughtered and blood samples were collected. Serum was used for analysis of aspartate aminotransferase (S-ASAT), alanine aminotransferase (S-ALAT), alkaline phosphatase (S-ALP), S-cholesterol, S-triglycerides, S-fructosamine, S-urea, S-total protein, and for electrophoretic fractionation of serum proteins, corticosteroid-binding globulin (CBG), cortisol, progesterone, thyroxine (T4) and free T4. There were no significant differences between groups in embryonic survival or in number of viable fetuses after treatment with glucocorticoids. The activity of S-ALP was lower (p < 0.05) in Group 4 than in Group 3 (0.5 vs 1.2 mukat/l). Group 4 had higher (p < 0.05) levels of S-triglycerides (1.17 vs 0.73 mmol/l), S-cholesterol (5.4 vs 2.7 mmol/l), S-total protein (110.5 vs 93.3 g/l), S-albumin (56.3 vs 43.3 g/l) and alpha 2-globulin concentrations (18.0 vs 14.3 g/l) than Group 3. The hydrocortisone-treated gilts had lower (p < 0.05) CBG (6.8 vs 21.3 nmol/l) and beta 1-globulin (3.25 vs 5.0 g/l) concentrations than the oil-treated ones. Concentrations of T4 were lower (p < 0.05) in Groups 2 (61.3 nmo/l) and 4 (49.0 nmol/l) compared with control Groups 1 and 3 (88.2 and 97.0 nmol/l, respectively). Overall, the treatment of early pregnant gilts with hydrocortisone acetate resulted in decreased levels of S-ALP, CBG, beta 1-globulin and T4, and in increased levels of S-cholesterol, S-triglycerides, S-total protein, S-albumin and alpha 2-globulin. The only effect of dexamethasone was a lowering of T4. There were no differences in free T4, S-fructosamine or S-urea between controls and treatments. Furthermore a negative correlation between triglycerides concentrations and the number of embryos (r = -0.76, p < 0.05) was found in control untreated and oil-treated pregnant gilts.

Effects of melatonin infused into the III ventricle on prolactin, beta-endorphin and luteotropin secretion in ewes during the different stages of the reproductive cycle.

Secretion of all the pituitary hormones undergoes marked circadian and seasonal changes. The rhythmicity of these changes is controlled by the circadian pacemaker system and the pineal gland transmitting daylength information to the neuroendocrine axis via the secretion of melatonin. This article presents data on the effects of the short-term melatonin administration into the third brain ventricle on prolactin, beta-endorphin and luteotropin secretion in ewes kept under the increasing and decreasing daylength conditions. Additional emphasis is given to dopamine and LHRH release in the mediobasal hypothalamus under the melatonin treatment by the push-pull method. The long-term and short-term actions of melatonin on the hormonal status in ewes is also discussed.

Calcium channel blocker prevents stress-induced activation of renin and aldosterone in conscious pig.

A considerable amount of data suggest the involvement of calcium-mediated processes in the activation of the renin-angiotensin-aldosterone (RAA) cascade. To investigate the effect of calcium-channel inhibition on the RAA system, we studied 21 conscious pigs. Blood renin and aldosterone levels increased by subjecting animals to 24 hours of immobilization stress. Renin and aldosterone levels were repeatedly measured by radio-immunoassay in blood samples taken periodically over 24 hours from a chronically implanted arterial cannula. Pretreatment of the animals (N = 11) with nisoldipine, 2 x 20 mg p.o. daily for 2 days before and on the day of immobilization, transiently attenuated the stress-induced increase of plasma renin activity and completely prevented the rise of aldosterone, as compared to nontreated controls (N = 10). The finding that nisoldipine suppresses RAA activation induced by a nonpharmacologic stimulus in the conscious intact animal may have clinical implications.

Stress-induced injury of pig myocardium is accompanied by increased lipid peroxidation and depletion of mitochondrial ATP.

The aim of the study was to investigate the mechanisms of myocardial lesions induced by stress in conscious, intact pigs. The animals were subjected to 24 h immobilization, controls were kept in normal conditions. The pigs were killed by electric shock and exsanguination. Lipid peroxidation products: malondialdehyde (MDA), conjugated dienes (CDB), fluorescent end products (RF) and adenine nucleotides (ATP, ADP) were measured in the left ventricular myocardium which was also subjected to histoenzymatic and electron microscopic examination. In stressed animals as opposed to the control group, alterations in ultrastructure and diminution of mitochondrial ATP have been found, together with augmented formation of MDA and CDB reflecting increased free radical generation. These changes may be the component of stress-induced myocardial injury.

Effect of nisoldipine on stress-induced myocardial damage in the conscious pig.

1. The effect of the calcium channel blocker nisoldipine on the myocardial content of lipid peroxidation products (malondialdehyde (MDA), conjugated double bonds (CDB), fluorescent end-products (RF) and mitochondrial adenine nucleotides) was investigated in conscious pigs (n = 14) subjected to 24 h of immobilization stress. Histoenzymatic and electron microscopic studies of the myocardium were also performed. Nisoldipine was given orally in a twice daily dose of 20 mg for 2 days before and on the day of the experiment. Results were compared with those obtained in immobilized untreated pigs (n = 10) and in non-stressed treated controls (n = 8). 2. Pretreatment with nisoldipine significantly attenuated stress-induced increase in myocardial contents of CDB and RF and prevented decline of mitochondrial adenine nucleotides. Stress-induced myocardial histoenzymatic changes (decrease of succinic dehydrogenase, ATPase, acid phosphatase activity) and ultrastructural alterations (mitochondrial damage, lysis of myofibrils, dilatation of sarcoplasmic reticulum and endothelial swelling) were also diminished. 3. It is concluded that treatment with a Ca2(+)-antagonist is beneficial to the heart exposed to environmental stress.

Peripheral plasma prolactin concentrations during oestrous cycles in different types of primitive gilt.

Plasma prolactin concentrations were determined by radioimmunoassay during oestrous cycles and around the time of oestrus in different types of primitive gilts: Vietnamese, Zlotnicka and wild-boar X domestic pig hybrids. The animals were bled without stress from an indwelling arterial catheter. The following results were obtained: (1) in all gilts the main prolactin peak was observed at Day 15 or 16 of the oestrous cycle; (2) Vietnamese and hybrid gilts showed a second smaller prolactin surge after (Day 2) or before (Day 17) oestrus; (3) base levels of prolactin during the oestrous cycle were 14.8 +/- 0.93 ng/ml (Vietnamese gilts), 13.2 +/- 1.05 ng/ml (Zlotnicka gilts) and 15.6 +/- 2.01 ng/ml (hybrid gilts). The 15-16-day prolactin peaks reached maximum values of 36.4, 43.4 and 56.5 ng/ml respectively.

[Effect of pregnant mare serum on bovine luteal tissue]. / Ucinnost séra brezích klisen na luteální tkáne skotu.

Trials were conducted with four heifers and twelve dairy cows to study the luteotropic effect of the serum of pregnant mares administered at the rates of 2000 to 3000 units. The effect of serum gonadotropin (PMSG) on ovarian activity was compared with the ovarian activity of control animals and ovarian activity after the administration of hypophyseal hormones. The highest progesterone concentration of PMSG-treated cows ranged from 41.9 to 74.3 ng per ml of milk; in the control group this range was between 32.2 and 43.1 ng per ml and in the group of gonadotropin-treated cows between 31.7 and 44.8 ng per ml. In heifers the progesterone levels increased from 3.56 to 4.58 ng per ml of plasma within 24 hours from the administration of PMSG. After 48 hours from administration the average progesterone concentration increased to 11.02 ng per ml of plasma. The animals which did not respond to PMSG administration by the growth of follicles exhibited an increased secretion of progesterone. The administration of hypophyseal hormones in the luteal stage of sexual cycle did not exert any significant influence on progesterone concentration, but stimulated the growth of follicles.

[Adrenocortical function in myasthenia pseudoparalytica]. / Badania nad czynnoscia kory nadnerczy w miastenii rzekomoporaznej

In a group of 31 women with myasthenia the urinary excretion of 17-ketosteroids and their fractions (11-deoxy-17-ketosteroids, dehydroisoandrosterone) and 17-hydroxycorticosterone were determined. A statistically significant decrease was observed in the excretion of these substances. Besides that, the plasma cortisol level was determined in these patients and the results obtained in the whole group were at the lower normal range of the laboratory and in 40% of cases they were significantly lower (p less than 0.01). The obtained results may suggest that adrenocortical hypofunction exists in many cases of myasthenia. The pertinent literature is reviewed.

Plasma levels of LH and prolactin in cows following enucleation of the persistent corpus luteum.

LH and prolactin were measured in plasma of 5 cows before and after enucleation of the persistent corpus luteum. Two cows returned to normal oestrous cycles following enucleation and did not differ from normal, cycling cows, with respect to hormone levels. Three other ones developed follicular cysts following enucleation and had decreased levels of LH and prolaction as compared with cycling cows. On the day of enucleation the fluctuations in LH-levels were higher than either before or after enucleation.

Effect of castration, testosterone treatment and hereditary sterility on prostaglandin concentration in the male reproductive system of mice.

The concentration of prostaglandins E and F in different parts of the male reproductive system of CD-1 and dwarf mice was measured by radioimmunoassay. In intact CD-1 mice, the vas deferens contained a significantly higher concentration of PGE and PGF than was found in the epididymis and in the seminal vesicles. All tissues studied had more PGE than PGF. Castration resulted in increased PG concentrations in both the epididymis and the seminal vesicles and decreased PG concentrations in the vas deferens. There was also a differential response of PGE and PGF in the epididymis of genetically sterile dwarf mice (dw/dw) were significantly higher than those observed in their normal littermates ((PLUS)). A reversed PGE/PGF ratio was found in the mates. The results indicate that testicular androgens affect the levels of PGE and PGF in the reproductive system of male mice. The physiological role of PGs in male reproductive functions has not been established, but there is a suggestion that PH have a role in controlling the transit of spermatozoa through the epididymis and vas deferens.