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Effect of estrogen administration on the induction of the plasma prolactin afternoon surge and on anterior pituitary prolactin concentration extracted at different pHs.

Ovariectomized (OVX) rats were injected with various doses of polyestradiol phosphate (PEP); the anterior pituitary (AP) prolactin (PRL) concentration and the plasma afternoon surge of PRL were observed 1 week later by radioimmunoassay. AP PRL was extracted using carbonate and phosphate buffers at either pH 7.6 or 10.6. The AP concentration of PRL was greater when the AP was extracted with buffers at pH 10.6 and the phosphate buffer was the most efficient. The concentration of PRL in the AP more closely reflected the magnitude of the estrogen-induced afternoon surge when the AP was extracted at pH 10.6 and this was especially so when the higher levels of estrogen were administered.

Animals↗

Prolactin, estriol and progesterone levels in frequent blood samples throughout normal pregnancy (lack of prolactin pulsatile secretion).

The aim of this paper was to study the episodic fluctuations of circulating prolactin (PRL), estriol (E3) and progesterone (P4) concentrations throughout pregnancy. We examined 24 pregnant women; 21 were between the 28th and 40th week of gestation, and the other 3 in the 12th, 16th and 20th week of gestation. Blood samples were drawn every 5 min for half an hour, and every 15 min for one and a half hour. Blood samples were taken in two and three different weeks of gestation in 11 and 2 of the cases, respectively. Two normal non-pregnant women were also studied and used as controls. PRL, E3 and P4 were determined by radioimmunoassay in all the samples. The coefficients of variation of PRL values were 40 and 22.6%, respectively, in the two control women, 8, 12 and 9.8% in the pregnant women studied at the 12th, 16th and 20th week of gestation, respectively, while in the 21 cases studied during the third trimester the coefficient of variation was 8 +/- 3% (mean +/- SD). The coefficients of variation of the values obtained for E3 and P4 in women studied in he third trimester were 26 +/- 15 and 16 +/- 6% (Mean +/- SD), respectively. There was an increase in the average concentration of the three hormones in all the cases at two or three different weeks. We can conclude that E3 and P4 have a pulsatile secretion pattern throughout pregnancy, and that PRL looses its pulsatile secretion as from an early gestational age. Our results suggested that central mechanisms regulating PRL episodic fluctuations were altered during pregnancy.

Estriol↗

The duration of prolactin secretory bursts from the pituitary is independent from both prolactin and gonadal steroid plasma levels in women and in men.

The intrinsic secretory characteristics of prolactin (PRL) have been investigated using newly developed algorhythms for instantaneous secretory rate (ISR) computation. PRL secretory rate, its intrinsic pulsatile characteristics and their possible dependance from gonadal steroids were investigated in five groups of subjects: a) 11 women during the follicular and luteal phase of the same menstrual cycle; b) 5 healthy postmenopausal women; c) 6 women affected by functional hyperprolactinemia; d) 5 normal men; e) 4 agonadal subjects before and during testosterone replacement therapy. All subjects underwent a 6 hours pulsatility study, from 08:00 to 14:00, sampling every 10 minutes. PRL plasma concentrations were determined using a RIA system and the presence of PRL secretory pulses was evaluated with program DETECT, both on plasma time series and after ISR computation. A distinct PRL episodic release was observed in all groups (follicular phase: 5.5 +/- 0.5, luteal phase: 6.5 +/- 0.6, postmenopause: 5 +/- 1, hyperprolactinemic women: 4.2 +/- 0.8, men: 4.8 +/- 0.4, agonadal before testosterone: 6 +/- 1, agonadal during testosterone administration: 5.3 +/- 0.3 peaks/6h), but mainly the computation of ISR allowed to demonstrate that the duration of the lactotropes secretory events was constant in all groups studied. PRL secretory bursts duration ranged between 23.1 +/- 1.8 and 25.4 +/- 2.5 minutes independently both on PRL or on sex steroid plasma levels. In conclusion, the present report shows that in different physiological conditions the intrinsic secretory bursts from lactotropes are constant in duration independently from the functional state, sex and the steroid hormone levels.

Adult↗

Diethylstilbestrol increases the density of prolactin cells in male mouse pituitary by inducing proliferation of prolactin cells and transdifferentiation of gonadotropic cells.

Diethylstilbestrol (DES) has been implicated in mammalian abnormalities. We examined the effects of DES on follicle-stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) cells in the pituitaries of male mice treated with various doses of DES for 20 days. DES reduced the density of FSH and LH cells in a dose-dependent manner, but increased that of PRL cells. When the expression of estrogen receptor (ER) alpha and beta was assessed, an induction of ERbeta by DES was found predominantly in PRL cells. However, since these effects were abolished in ERalpha knockout mice, DES appears to act primarily through ERalpha. When the expression of Ki-67 and Pit-1 in PRL cells was examined at various time-points after DES treatment, some PRL cells became Ki-67 positive at 10-15 days, and Pit-1-positive cells were increased at 5-15 days. Furthermore, some FSH and LH cells became Pit-1 positive, and co-localized with PRL at 5-10 days. Our results indicate that DES increases PRL cells by inducing proliferation of PRL cells and transdifferentiation of FSH/LH cells to PRL cells.

Animals↗

Prolactin in patients with major depressive disorder and in healthy subjects. I. Cross-sectional study of basal and post-TRH and postdexamethasone prolactin levels.

Prolactin (PRL) levels were investigated in patients with major depressive illness and in healthy subjects. Basal and postdexamethasone levels were measured in 27 patients, and levels after thyrotropin-releasing hormone (TRH) stimulation (delta PRL) were measured in 22 patients. Basal and delta PRL were also determined in 64 age- and sex-matched healthy subjects. Both basal and postdexamethasone PRL levels were normal in depressed patients, with the postdexamethasone levels in particular showing no correlation to postdexamethasone cortisol concentrations. One milligram oral dexamethasone did not influence 4:00 PM PRL levels in 15 healthy subjects. delta PRL was significantly elevated in both male and female patients. These increases were not correlated with severity of illness and are difficult to interpret owing to the complexity of the PRL regulatory system. No significant correlations were found between basal or post-TRH PRL and cortisol, thyroid-stimulating hormone (TSH), thyroid hormones, gonadotropins, or estradiol in the patients. However, surprisingly significant positive correlations between basal PRL and basal cortisol, T4 and reverse T3 occurred in healthy subjects. It is as yet unclear how this finding can be explained and what relevance it has. Women tended to have higher basal PRL concentrations than men, but the difference was not significant in either group. delta PRL was significantly higher in women than in men in both patients and controls. No significant influence of age was found.

Adult↗

Lack of the suckling-induced prolactin release in homozygous Brattleboro rats: the vasopressin-neurophysin-glycopeptide precursor may play a role in prolactin release.

Suckling stimulus did not induce significant release of prolactin (PRL) in lactating homozygous Brattleboro rats, whereas it did it in heterozygous animals. Daily treatment of homozygous rats with vasopressin partly restored the PRL response to suckling. Findings suggest that vasopressin-neurophysin-glycopeptide precursor missing in homozygous Brattleboro rats may play a role in suckling-induced PRL release.

Animals↗

The influence of thyrotropin releasing hormone on in vivo prolactin release and in vitro prolactin, luteinizing hormone, and growth hormone release from dispersed pituitary cells of the young turkey (Meleagris gallopavo).

Intravenous administration of 0.025, 0.25, or 2.5 micrograms/kg thyrotropin releasing hormone (TRH) to 4-week-old female turkeys induced a dose-dependent increase (P = 0.004) in serum prolactin (PRL) 15 min post-treatment. Dispersed anterior pituitary cell cultures were utilized to determine the effect of TRH on cellular release of PRL, luteinizing hormone (LH), and growth hormone (GH). In the first experiment, cells from 13-week-old male turkeys were initially incubated for 24 hr in Medium 199 (M-199) plus 10% turkey serum and then placed in M-199 plus 10(-10) to 10(-4) M TRH for 5 hr. Incubation with TRH produced no change in PRL release from that of spontaneous release (P = 0.854). However, 10(-5) and 10(-4) M TRH induced LH release (P less than 0.0001). The TRH-induced GH response was parabolic (P less than 0.0001), with the maximal release at 10(-8) M. The second experiment, utilizing pituitary cells from 7-week-old females, studied these responses on 3, 5, and 7 days of monolayer incubation. TRH failed to induce a PRL release in all tests (P greater than 0.162), although hypothalamic extract induced a large release (P less than 0.0001) of PRL each time. Both 10(-6) and 10(-4) M TRH induced a LH release on Day 3 while only 10(-4) M did so on Day 5, and none of the doses elicited a release on Day 7. The parabolic GH response generally persisted in all tests.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Is the pineal involved in the stimulatory influence of prolactin on tail regeneration in lizards? Studies with exogenous prolactin in lizards exposed to continuous darkness.

We have recently demonstrated that continuous light stimulates tail regeneration in the gekkonid lizard, Hemidactylus flaviviridis, whereas continuous darkness and pinealectomy depress regeneration. As a sequel to this observation, the effect of exogenous ovine prolactin (oPRL) on tail regeneration in normal (NL) and pinealectomized (PX) Hemidactylus exposed to continuous darkness (L:D 0:24) during the monsoon season was investigated. Once-daily intraperitoneal (ip) injections of 500 micrograms/kg oPRL were administered to a group of NL lizards, a group of PX lizards, and a group of sham-PX (sPX) lizards, 5 days prior to tail autotomy and 50 days afterward. Three groups--PX, sPX, and NL animals--served as the controls and received once-daily ip injections of 0.6% saline. Our observations show that initiation of regeneration, daily growth rate, total length of new growth (regenerate) at the end of regeneration, and total percentage replacement of lost (autotomized) tail were all significantly enhanced in oPRL-treated NL lizards as compared with their saline-treated (NL, PX, and sPX) and oPRL-treated (PX and sPX) counterparts. It is suggested that PRL may be the active factor that speeds up the rate of tail regeneration in lacertilians. The role of the pineal organ in vertebrate photoreception and the possibility that in Hemidactylus the presence of an intact pineal is somehow linked with the favorable influence of PRL on tail regeneration are discussed.

Animals↗

Immunocytochemistry and immunoblotting of avian prolactins using polyclonal and monoclonal antibodies toward a synthetic fragment of chicken prolactin.

A major obstacle in the production of specific antibodies toward chicken prolactin (PRL) has been overcome by mimicking a putative epitope of the molecule using the synthetic decapeptide Lys-chPRL 59-67. This peptide represents the highest hydrophilicity peak of the amino acid sequence of chPRL that was recently derived from the nucleotide sequence. Polyclonal mouse antisera against the fragment specifically recognized the lactotropes in the cephalic lobe of the chicken pars distalis as illustrated by immunocytochemical double staining experiments. Monoclonal antibody production yielded antibodies that specifically labeled purified turkey PRL upon SDS-PAGE separation and immunoblotting. Turkey and chicken PRL showed a very similar polymorphism with respect to their apparent molecular weights, including the occurrence of a glycosylated variant of chicken PRL. The monoclonal antibodies were finally used to demonstrate the presence of PRL-like immunoreactivity both in the pituitary gland and in the brain of the quail. In the brain, immunoreactive neurons were in the nucleus accumbens and in the lateral parts of the ventro-medial hypothalamus, partly similar to those described in the rat.

Amino Acid Sequence↗

Paradoxical effects of oxytocin and vasopressin on basal prolactin secretion and the estrogen-induced prolactin surge.

The roles of oxytocin (OT) and vasopressin (AVP) on both basal and estrogen-induced prolactin (PRL) secretion were examined. Adult female Sprague-Dawley rats that were ovariectomized for 3 weeks and received estrogen treatment for 1 week were used. Intravenous administration of hormones and serial blood sampling were accomplished through indwelling intraatrial catheters which were implanted two days before. Plasma PRL levels were measured by radioimmunoassay. Oxytocin at a dose of 20 micrograms/rat stimulated a moderate PRL release in the morning and lower doses (5 and 10 micrograms) were without effect. Vasopressin was most effective at a dose of 5 micrograms/rat in stimulating PRL release, while consecutive injections of higher doses (10 and 20 micrograms) were less effective. In contrast, TRH, ranging from 1 to 8 micrograms/rat, induced a dose-dependent increases in PRL secretion. Using the effective dosages determined from the morning studies, repeated injections of either OT, AVP or their specific antagonists MPOMeOVT [( 1-(beta-mercapto-beta, beta-cyclopentamethylene propanoic acid), 2-(O-methyl)tyrosine, 8-ornithine]-vasotocin) and d (CH2)5Tyr(Me)AVP ([1-(beta-mercapto-beta, beta-cyclo-pentamethylene propionic acid), 2-(O-methyl)tyrosine, 8-arginine]-vasopressin), were given hourly between 1300 to 1800 h and blood samples were obtained hourly from 1100 to 1900 h. It was found that either OT or AVP significantly reduced the afternoon PRL surge, while their antagonists were not as effective. When OT or AVP were administered together with their specific antagonists, the inhibitory effects of either hormone on PRL surge were reversed. Thus it is concluded that both OT and AVP assume a non-specific stress-like effect on PRL release, in which basal secretion is stimulated and surge secretion is inhibited.

Animals↗

Prolactin messenger ribonucleic acid concentrations throughout the ovine estrous cycle: assessment relative to prolactin serum and pituitary amounts.

Prolactin (PRL) mRNA concentrations were assessed by nucleic acid hybridization assays in pituitaries of ewes representing the defined stages of the ovine estrous cycle. Concomitantly, pituitary and serum PRL concentrations were measured in these ewes using radioimmunoassays. It was observed that PRL serum, pituitary and mRNA concentrations tended to increase near the time of the gonadotropin preovulatory surge, particularly between 24 hrs before behavioral estrus to 5 hours after estrus. However, the changes in PRL mRNA, serum and pituitary concentrations were shown not to be statistically significant. These data suggest that PRL production during the sheep estrous cycle is maintained without dramatic changes in synthesis or secretion.

Animals↗

Effects of haloperidol and prolactin secreting tumors on cerebrospinal fluid concentrations of prolactin in the female rat.

Studies were conducted to determine the effects of acute and chronic elevations in prolactin (PRL) secretion on serum and cerebrospinal fluid (CSF) PRL concentrations in the female rat. Young female rats showed a dose-dependent increase in serum and CSF PRL in response to haloperidol. A time-course evaluation of serum and CSF PRL levels after haloperidol indicated that serum PRL concentrations increased markedly by 30 min and declined thereafter; while CSF PRL increased more slowly, peaking at 2 to 8 h. In young rats with basal serum PRL levels, CSF PRL was maintained at 0.8 to 2.1% of serum PRL levels. During acute hyperprolactinemia, the CSF to serum PRL ratio increased to about 4%. During chronic severe hyperprolactinemia, induced by the growth of a MtT.W15 tumor, CSF PRL concentrations increased to 75 ng/ml, but this represented only 1.5% of serum PRL concentrations. Collectively, these data indicate that the blood-brain barrier effectively limits access to the brain of circulating PRL.

Analysis of Variance↗

Prolactin and growth hormone responses to dermorphin in patients with prolactin-secreting pituitary adenoma.

We have recently shown that dermorphin (D), a new potent opioid peptide (H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2) stimulates prolactin (PRL) and growth hormone (GH) secretion in humans. In 11 patients with a PRL-secreting pituitary adenoma (eight microprolactinomas and three macroprolactinomas with suprasellar extension), diagnosed by pituitary dynamic function tests, and radiological evidence with confirmation at surgery, the PRL and GH responses to D were studied to evaluate the effect of pathological hyperprolactinemia on the opioid-induced secretion of GH and PRL. No PRL response to D was observed in all 11 patients. Plasma GH increased after D in all patients, except in three patients bearing a macroprolactinoma. This study shows that the effect of D on PRL and GH secretion can be dissociated in patients with PRL-secreting pituitary adenoma, perhaps for a different derangement in the hypothalamic-pituitary mechanism(s) underlying the opioidergic regulation of GH and PRL secretion. In addition our data indicate that D can be employed as a useful opioid probe in humans.

Adenoma↗

Cysteamine, zinc, and thiols modify detectability of rat pituitary prolactin: a comparison with effects on bovine prolactin suggests differences in hormone storage.

Little is known about the structure of prolactin (PRL) within secretory granules. Evidence from our previous studies in bovine tissue preparations suggests that control of secretion may reside, in part, in the conversion of storage hormone to releasable PRL. The conversion can be monitored by measuring changes in immunodetectability since the oligomeric, storage form is poorly recognized by antisera raised against monomeric PRL. Since many investigators use rats to study the secretory process and changes in detectability of rat pituitary PRL occur during lactation ("depletion-transformation"), we undertook the present immunodetectability studies to gain insight into the storage structure of rat (r) PRL. Cysteamine and zinc inhibited tissue PRL immunoassayability in male rat pituitary homogenates and also in partially purified secretory granules as they had inhibited bovine (b) PRL; however, zinc inhibited the rodent hormone less potently than the bovine. In vitro incubation of rat tissue samples without additions resulted in increases in rPRL detectability of up to 84% after 180 minutes; such incubation of bovine samples had no significant effect. A striking additional difference between the species was that exposure to reduced glutathione (GSH), cysteine, homocysteine, mercaptoethanol, and dithiothreitol inhibited rPRL by up to 44%. This compared to thiol stimulation of bPRL by as much as 450%. The inhibitory GSH effect on rPRL was abolished when 0.5% sodium dodecyl sulfate (SDS) was included; in contrast, the stimulatory GSH effect on bPRL did not change with added SDS. SDS alone had no effect on rat homogenate PRL, and only increased rat granule rPRL by 23% compared to its ability to increase bPRL assayability by 44%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of endogenous prolactin on the luteinizing hormone stimulation of testicular steroidogenesis and the role of prolactin in adult male rats.

The role of endogenous prolactin (PRL) in the control of testosterone (T) secretion and T responses to LH treatment was evaluated in adult male rats. Rats were actively immunized three times against ovine PRL in Freund's adjuvant-saline mixture (PRL-IMM rats), and control rats were treated with adjuvant-saline mixture (ADJ-CON rats). On day 110 after initial immunization, rats in each of these two groups were divided into three subgroups. Rats in subgroups 1 and 2 were injected with saline while those in subgroup 3 received 200 micrograms ovine PRL in saline, twice a day for a total of 7 injections. On day 113, the seventh injection was given 3 h before the termination of the experiment. On the same day, 2.5 h before the rats were sacrificed, rats in subgroups 1 and 3 were treated with saline; animals in subgroup 2 received 25 micrograms ovine LH in saline. Blood samples were obtained throughout the study, and sera were used for measurement of PRL antibodies, gonadotropins, progesterone (P), and T. PRL antibodies were detected in the sera of all rats actively immunized with PRL. Administration of PRL increased serum T levels in ADJ-CON rats, and this effect was eliminated in rats actively immunized against PRL. LH treatment significantly increased serum T levels in ADJ-CON rats. In PRL-IMM rats, this increase was attenuated while circulating P concentrations were elevated. These data demonstrate that PRL treatment can increase T secretion and that endogenous PRL is required for the complete expression of the stimulatory action of LH on T secretion in adult male rats.

Animals↗

Prolactin cells and plasma prolactin levels in the aging male Wistar rat: an immunocytochemical and biochemical study.

Non-neoplastic pituitary glands of 12 age-groups of 10 male Wistar rats (subpopulation 1) and of 228 rats with age-related disorders (population 2) were studied. For subpopulation 1 a positive correlation was found between body weight, pars distalis volume and total prolactin (PRL)-cell volume. These values increased during the first half of the life span and decreased to values similar to that found in young rats, after the 50% survival age was reached. Age-related changes were found with respect to the distribution of the PRL cells (from homogeneous into non-homogeneous) and to their ultrastructural features (PRL cells with polymorphic granules were substituted by cells with round granules). Pituitary glands of disordered aged rats (subpopulation 2) showed a similar non-homogeneous distribution of PRL cells. Plasma PRL levels of both subpopulations were similar. No age-related increase of plasma PRL level was found in rats with non-neoplastic pituitary glands.

Aging↗

The effect of chronic apomorphine treatment on the ultrastructure of the prolactin cells and on plasma prolactin levels in young and aged male Wistar rats.

Effects of two doses of apomorphine on the plasma prolactin (PRL) levels and on the ultrastructure of PRL cells in young and aged male Wistar rats were investigated. In young and aged control rats no significant differences were found between the plasma PRL levels. Immunocytochemical staining with anti-r-PRL revealed significant differences between young and aged control rats; in young rats the number of PRL cells with polymorphic granules exceeded the number of cells with round granules, whereas in aged rats almost exclusively cells with round granules were found. In young rats, chronic treatment with a low dose (0.01 mg/kg/day) of apomorphine did not result in a significant change in plasma PRL level or cell morphology. However, high dose (0.25 mg/kg/day) of apomorphine resulted in a significant decrease in plasma PRL levels, a decrease of number of cells with polymorphic granules and an increase of cells with round granules. The occurrence of PRL cells with round granules and plasma levels was negatively correlated. In aged rats, apomorphine (0.01 or 0.25 mg/kg/day) treatment did not affect plasma levels nor did it affect the distribution of the cell types. We conclude that in young rats PRL cells are sensitive to apomorphine and that their ultrastructure reflects a phase of the secretory cycle. In aged rats, the cells appear to have lost their sensitivity to apomorphine. The fact, that the distribution over the different cell types in control aged rats is similar to that of the apomorphine-treated young rats, suggests a strong influence of endogenous dopamine on PRL cell physiology in the aged rat.

Animals↗

In vivo studies on prolactin function in the female rat: divergent effects of treatment with bromergocryptine and antisera to rat prolactin.

The effect of suppression of prolactin (PRL) serum levels on early pregnancy and lactation in the rat by means of either bromergocryptine (BEC) or rabbit antiserum to rat PRL has been examined. BEC terminated early pregnancy when given on any day from 1 to 6 and significantly impaired lactation performance when injected during the lactation period. Antibodies against rat PRI, however, showed no effect on either early pregnancy or lactation, although the sera of the treated rats contained free PRL antibodies as evidence for sufficient dosage of antiserum. This finding can be explained in two ways: either the antibodies are not able to neutralize the biological activity of the bound PRL, or the effect of BEC is not mediated by PRL-suppression alone but requires a second component not provided by the specific antiserum.

Animals↗