Pituitary Prolactin mRNA Levels Research Articles

Mammary gland-specific regulation of GNRH and GNRH-receptor gene expression is likely part of a local autoregulatory system in female vizcachas (Rodentia: Chinchillidae)

In addition to key mammotrophic hormones such as the pituitary prolactin (PRL) and the ovarian steroids progesterone and estradiol, there are local factors that modulate the tissue dynamics of the mammary glands during pregnancy and lactation. By immunohistochemistry and RT-PCR, we found local transcription and translation of gonadotropin-releasing hormone (GNRH), GNRH receptor (GNRHR), PRL and PRL receptor (PRLR) in mammary glands of adult vizcachas during pregnancy and lactation. Both GNRH and GNRHR showed a lag between protein expression and gene transcription throughout the gestational period: while the highest transcription levels of these genes were recorded at early-pregnancy, the epithelial immunoexpressions of both showed their maximum during lactation. RIA results corroborated the presence of GNRH in mammary glands at all the analyzed stages and confirmed the maximum amount of this peptide in the lactating group. Significant amounts of GNRH were detected in milk samples as well. Conversely, PRL and PRLR shared similar protein and gene expression profiles, all exhibiting maximum values during lactation. GNRH peptide content in mammary glands of females with sulpiride-induced hyperprolactinemia (HP) was significantly lower than that of control females (CT). Although PRL mRNA levels remained unchanged, there was a marked increase in theα-lactalbumin (LALBA) transcription in mammary glands of HP- vs CT-females. These results suggest that after targeting mammary glands, PRL stimulates the expression of milk protein genes, but also, tempers the local expression of GNRH. Mammary gland-explantssupplemented with a GNRH analogue (GN-explants) had no differences in terms of PRLR orLALBA transcription levels compared to CT-explants, so the mammary PRLR signaling would not appear to be modulated by GNRH. Yet, mRNA expression levels of both GNRH and the GNRHR-downstream factor, EGR1, were significantly higher in GN-explants compared to that of CT which would point to a GNRH-positive feedback mechanism. In summary, the local coupled expression of GNRH, GNRHR and EGR1 in the mammary gland throughout pregnancy of vizcachas, the PRL-dependent mammary GNRH secretion as well as the GNRH positive feedback on its own transcription suggest an autocrine-paracrine regulatory mechanism and propose an active role for GNRH in mammary gland tissue remodeling.

Does silvering or 11-ketotestosterone affect osmoregulatory ability in the New Zealand short-finned eel (Anguilla australis)?

Silvering has been associated with advancing osmoregulatory ability. Given the demonstrated role of 11-ketotestosterone (11KT) in mediating many of the silvering-related changes, we investigated the role of 11KT in driving this advanced osmoregulatory ability in the New Zealand short-finned eel (Anguilla australis). Yellow (non-migratory) eels with or without 11KT implants and blank-implanted silver (migratory) eels, either held in freshwater or subjected to seawater challenge, were sampled to determine serum [Na+] and [Cl-], pituitary prolactin mRNA levels, gill Na+/K+-ATPase activity and gill mRNA levels for Na+/K+-ATPase-α1 subunit and for Na+/K+/2Cl- co-transporter-1α-subunit. Developmental stage and 11KT treatment advanced the eels' osmoregulatory ability. Thus, serum [Na+] and [Cl-] were affected by developmental stage and 11KT treatment upon seawater challenge. However, seawater challenge, not 11KT treatment or developmental stage, produced the strongest and the most consistent effects on A. australis osmoregulatory processes, inducing significant effects in all the relevant parameters we measured. In light of our results and in view of the eel's marine ancestry, we contend that A. australis, or freshwater eels in general, are highly tolerant and able to adapt quickly to changing salinities even at the yellow stage, which may preclude a critical need for an advanced osmoregulatory ability at silvering.

Underground water affects sexual behavior and gene expression of hormones related to reproduction in blue gourami males

This study examined the effect of underground water on reproduction- and growth-related hormones in blue gourami males under non-reproductive and reproductive conditions. An increase in the percentage of males building nests under the highest percentage of underground water were compared to fish that maintained a lower percentage of underground water in the first two days. The % Gonado-somatic index (GSI) of males building nests was higher than non- reproductively active males in water containing the lowest concentration of underground water. In non- reproductively active males, brain gonadotropin releasing hormone 1 (GnRH1) and pituitary β subunit of gonadotropins (GtHs) and prolactin (PRL) mRNA levels were significantly higher in males maintained in underground water. In reproductively active males, mRNA levels of brain GnRH1, gonadotropin releasing hormone 3 (GnRH3) and pituitary PRL mRNA levels were significantly higher than males maintained in underground water. Thus, it is suggested that underground water with high salinity and conductivity levels affects the gene expression of repro- duction-related hormones; in reproductively active males, it shortened the duration of nest-building by blue gourami males.

Prolactin-releasing peptide, food intake, and hydromineral balance in goldfish

A potential role for prolactin-releasing peptide (PrRP) in appetite regulation and hydromineral balance in goldfish was examined. PrRP was found to be expressed in discrete regions of the goldfish brain, in particular, the hypothalamus. Intraperitoneal (IP) or intracerebroventricular administration of PrRP had dose-dependent effects to suppress food intake in goldfish. Hypothalamic PrRP mRNA expression significantly increased after feeding, as well as after 7 days of food deprivation. Refeeding fish after 7 days food deprivation did not result in a postprandial increase in PrRP mRNA expression. These data suggest an anorexigenic role for PrRP in the short term around a scheduled meal time, but not over the longer term. IP injection of PrRP significantly increased pituitary prolactin (PRL) mRNA levels, suggesting involvement in the regulation of lactotroph activity. Acclimating goldfish to an ion-poor environment decreased serum osmolality and increased PrRP and PRL mRNA levels, providing evidence for PrRP involvement in hydromineral balance through its actions on lactotrophs. Acclimation to ion-poor water diminished the anorexigenic properties of PrRP in goldfish, indicating that a role for PrRP in goldfish satiation is counterbalanced by alternate systemic needs (i.e., osmoregulatory). This was further supported by an ability to reinstate the anorexigenic actions of PrRP in fish acclimated to ion-poor water by feeding a salt-rich diet. These studies provide evidence that PrRP is involved in regulating appetite and hydromineral balance in fish, and that the degree of involvement in either process varies according to overall systemic needs in response to environmental conditions.

Salinity preference of silvering Japanese eel Anguilla japonica: evidence from pituitary prolactin mRNA levels and otolith Sr:Ca ratios

The salinity preference of Japanese eel Anguilla japonica during silvering was validated from the viewpoints of physiological ecology by examining the pituitary prolactin (PRL) mRNA expression and the otolith strontium:calcium (Sr:Ca) ratios of the eels of different sexual maturity stages collected in an estuary of Taiwan. The PRL possessed a putative signal peptide of 24 amino acids and a mature peptide of 185 amino acids. RT-PCR and real-time quantitative PCR analyses indicated that the transcript levels of PRL mRNA significantly decreased from yellow to silver stage. On the contrary, the Sr:Ca ratios at the otolith edge of silver eels, which recorded their latest experience of salinity history, significantly increased. The increase in otolith Sr:Ca ratios and the decrease in PRL mRNA expression are consistent with the timing of migration from freshwater to seawater during silvering of the eel.

Temperature-dependent prolactin secretion and reproductive biology of the newt Triturus carnifex Laur

The effects of temperature on pituitary prolactin (PRL) gene expression and peripheral levels were studied in both male and female newts obtained from wild conditions during reproductive (Experiment I) and nonreproductive (Experiment II) periods; moreover, changes in parameters related to reproductive function are also described. Male and female newts were taken from a pond in February (Experiment I, reproductive period) and maintained for 1 month in tanks at 4 and 18 °C. In male newts kept at 4 °C, increase of PRL mRNA in the pituitary and plasma PRL was found compared with that measured in those kept at 18 °C. The increase in PRL secretion was parallel to that of plasma androgens and related secondary sexual characteristics (SSC) in males and of plasma estradiol-17β and vitellogenin in females. On the contrary, in nonreproductive newts (Experiment II), taken from the field in May, no significant changes in plasma PRL, androgens, and SSC were found in those maintained at low temperature (4 °C), whereas low temperature significantly increased PRL mRNA expression in the male pituitary and PRL mRNA plus plasma PRL levels in females. These findings suggest that low temperature regulates PRL secretion in this urodele species, showing a sex- and season-related control mechanism; moreover, low temperature failed to influence the reproductive biology of newts taken from the field in May, after naturally occurring reproduction during winter months.

Influence of environmental salinity on messenger RNA levels of growth hormone, prolactin, and somatolactin in pituitary of the channel catfish (Ictalurus punctatus).

Pituitary growth hormone (GH), prolactin (PRL), and somatolactin (SL) messenger RNA levels in channel catfish (Ictalurus punctatus) were examined under various environmental and physiological conditions. Catfish were sampled following salinity challenge, during the winter (December) and spring or summer (April or July), and at different sizes (15-18 g, 620-664 g, and 956-1134 g). When catfish (956-1134 g) were transferred from freshwater to saline water containing 8 ppt NaCl, their plasma [Na(+)] increased significantly above values in the freshwater control group until they were transferred back to freshwater. Pituitary GH mRNA levels were low for the first 24 hours following transfer to saline water, but thereafter were significantly elevated above control values until the fish were transferred back to freshwater. Pituitary GH mRNA levels were highest in July and lowest in December. Growth hormone mRNA levels were also elevated in the size groups 15-18 g and 956-1134 g in July when compared with December values. Pituitary PRL mRNA levels increased for the first 24 hours following transfer to saline water (956-1134 g), but thereafter were significantly lower than control values until the fish were transferred back to freshwater. Pituitary PRL mRNA levels were highest in April and July and lowest in December, and were also elevated in the size groups 620-664 g and 956-1134 g. Pituitary SL mRNA levels were unaffected in catfish transferred to saline water; however, levels were significantly elevated in catfish of the 956-1134-g size group sampled in April when compared with December. These results suggest the involvement of GH in adaptation to brackish water and of PRL in adaptation to freshwater in the catfish, and seasonal and size-related differences in pituitary GH, PRL, and SL mRNA levels.

Chronic sustained stress increases levels of anterior pituitary prolactin mRNA

Our laboratory is investigating the effects of chronic stress on physiological, endocrine and behavioral measures, in order to elucidate the neuronal substrates for the pathophysiological consequences of stress in humans. In these studies, we have employed a rodent model of sustained stress in which rats are exposed to around-the-clock intermittent footshock that can be avoided or escaped by rats in the controllable stress group, but not by rats in the uncontrollable stress group. Each rat in the uncontrollable stress group is paired (yoked) to a rat in the controllable stress group such that the controllable stress group rat avoids or escapes shock for both rats. A third group of rats receives no shock (controls). We have previously reported that in male rats, plasma prolactin levels were elevated after 3 days of stress in both stress groups. In the present experiments we determined whether the increases in plasma prolactin were correlated with increases in anterior pituitary prolactin mRNA. In addition, we measured hormones and mRNA at three time points and we examined these responses in female as well as male rats. Adult male and female Sprague–Dawley rats were exposed to chronic stress for 1, 3 or 14 days. In unstressed control rats, levels of anterior pituitary prolactin mRNA were fivefold higher in female as compared to male rats. However, stress increased levels of anterior pituitary prolactin mRNA over baseline in both genders. After 1 day of stress, anterior pituitary prolactin mRNA levels increased in male and female rats belonging to both stress groups, with no significant difference seen between the uncontrollable vs. controllable stress groups. After 3 days of stress, anterior pituitary prolactin mRNA levels were even more elevated, and rats in the uncontrollable stress group had higher anterior pituitary prolactin mRNA levels than those in the controllable stress group. After 14 days of stress, there were no significant differences in control and stressed groups with respect to anterior pituitary prolactin mRNA. These data suggest that chronic sustained stress increases the synthesis of anterior pituitary prolactin mRNA during the first days of stress, and that levels return to prestress values sometime between 3 and 14 days of stress.

Cloning of Bullfrog Thyroid-Stimulating Hormone (TSH) β Subunit cDNA: Expression of TSHβ mRNA during Metamorphosis

A thyroid-stimulating hormone β subunit (TSHβ) cDNA encoding both signal peptide and mature TSHβ molecule was cloned from a cDNA library constructed from total RNA of the bullfrog (Rana catesbeiana) adenohypophysis. The bullfrog TSHβ mRNA was estimated by Northern blot analysis to be approximately 1 kb. The deduced amino acid sequence showed 40–61% homologies with the sequences of TSH β subunits of other vertebrates. Using the cDNA as a probe, we measured changes in mRNA expression in metamorphosing tadpoles of R. catesbeiana. The TSH β subunit mRNA level increased progressively throughout prometamorphic stages, reaching its maximum at the end of prometamorphosis. The maximum level was maintained throughout early and mid climax, declining at late climax. These results, together with previously obtained data on plasma prolactin and pituitary prolactin mRNA levels, as well as thyroid hormone levels, are discussed in relation to metamorphic changes occurring in the bullfrog larvae.

The relative roles of the hypothalamus and cortisol in the control of prolactin gene expression in the anterior pituitary of the sheep fetus.

The neuroendocrine control of prolactin synthesis and secretion before birth is not well understood. We have measured the changes in the level of prolactin mRNA in the anterior pituitary of the fetal sheep throughout the last 15 days of pregnancy (term = 147 +/- 3 days gestation). We have also investigated the effects of surgical disconnection of the fetal hypothalamus and pituitary (HPD) with or without long term cortisol infusion on pituitary prolactin mRNA levels and plasma prolactin concentrations in the late gestation sheep fetus. Prolactin mRNA levels were measured in anterior pituitaries collected from a series of fetal sheep (130-134 days, n = 6; 135-140 days, n = 6; 141-145 days, n = 6) in late gestation. HPD was carried out in ten fetal sheep at 105-115 days gestation and five intact fetal sheep were used as controls. In the HPD group, either saline (HPD + saline group, n = 5) or cortisol was infused (3.5 mg/24 h) for 5 days from 134-136 days gestation (HPD + cortisol group, n = 5). There was an increase in the ratio of prolactin mRNA: 18S rRNA in the fetal pituitary between 130-134 days (0.46 +/- 0.08, n = 6) and 135-140 days (1.27 +/- 0.17 n = 6) which was maintained after 141 days gestation, (1.27 +/- 0.11, n = 6). The mean prolactin mRNA: 18 S rRNA ratio was significantly higher (P < 0.05) in intact fetal sheep (1.41 +/- 0.16, n = 4) than in the HPD fetal sheep after either saline (0.54 +/- 0.14, n = 4) or cortisol (0.74 +/- 0.24, n = 5) administration. The mean plasma concentration of prolactin was also higher in the intact group (28.3 +/- 3.9 ng/ml) when compared with the HPD + saline group (8.0 +/- 3.3 ng/ml) or the HPD + cortisol group (5.6 +/- 1.9 ng/ml). We have demonstrated that there is a strong hypothalamic drive to prolactin synthesis and secretion in the fetus and that cortisol does not act directly at the fetal pituitary to stimulate prolactin synthesis and secretion in late gestation.

Estradiol Induces Vasoactive Intestinal Peptide and Prolactin Gene Expression in the Rat Anterior Pituitary Independently of Plasma Prolactin Levels

It is well established that estrogens are potent stimulators of prolactin (PRL) secretion. It has also been demonstrated that estradiol (E2) can increase the expression and the anterior pituitary levels of the vasoactive intestinal peptide (VIP), a peptide which also acts as a potent PRL-releasing factor. It thus remained unknown whether the effects on pituitary VIP were due to E2 itself or to E2-induced hyperprolactinemia (HPRL). In order to test this hypothesis, various plasma PRL levels were induced in rats either with ectopic pituitary grafts, PRL secreting tumours or E2 implants, and VIP mRNA expression in the anterior pituitary was measured by in situ hybridization and Northern blot analyses. Whereas decreases in VIP mRNA can be observed in pituitaries of rats with pure HPRL, a 6-fold increase in VIP mRNA can be seen in E2-treated rats. E2 increased both 1.0 and 1.7 Kb VIP mRNA species. The presence of the graft in E2-treated rats significantly reduced the increase in VIPmRNA observed following E2. The direct stimulation by E2 of VIP mRNA expression was further demonstrated by the fact that statistical analysis of the data indicated that both E2 and graft were acting independently of each other, and that a new selective antiestrogen, RU 58668, almost totally blocked the effect of E2. Moreover, under similar experimental conditions, pituitary PRL mRNA levels were reduced in the graft group and a marked up-regulation was observed similarly in both E2 and in E2 rats bearing ectopic grafts.(ABSTRACT TRUNCATED AT 250 WORDS)

Serotonergic involvement in the regulation of prolactin and vasoactive intestinal peptide mRNA expression in the rat anterior pituitary

These studies examined the contribution of serotonin (5-HT) to the control of prolactin (PRL) and vasoactive intestinal peptide (VIP) messenger RNA expression in rat anterior pituitary. Daily injection of rats with the biosynthetic precursor to serotonin, 5-hydroxytryptophan (5-HTP; 25 mg/kg q.i.d.), resulted on day 5 in a 50% increase in the expression of PRL mRNA in the pituitary while at the same time reducing the levels of both the 1.0 and 1.7kb VIP mRNA transcripts. Co-treatment of rats with 5-HTP plus the catecholamine biosynthesis inhibitor, α-methyl-tyrosine (α-MT; 150 mg/kg q.d. × 2 days), or the dopamine receptor antagonist haloperidol (1.25 mg/kg b.i.d. × 5 days), resulted in increases in pituitary PRL message levels that were greater than those observed with either anti-dopaminergic agent alone. In contrast, 5-HTP was unable to reverse the inhibition of PRL mRNA expression caused by treatment with the dopamine receptor agonist bromocriptine (2.5 mg/kg b.i.d. × 5 days). Neither α-MT, haloperidol nor bromocriptine had a significant effect on pituitary VIP mRNA expression. Administration of the direct-acting 5-HT receptor agonist quipazine (5 mg/kg, b.i.d.) for 14 consecutive days caused a significant increase in pituitary PRL mRNA levels on day 1 and reached a plateau of 90% above control levels on days 7 and 14. VIP mRNA levels rose significantly on day 1 of quipazine treatment but thereafter fell to a minimum of 22% (1.0 kb) and 52% (1.7 kb) of control by day 14. Both the 5-HT 1 receptor antagonist methiothepin (1 mg/kg t.i.d. × 3 days) and the 5-HT 2 receptor blocker ketanserin (8 mg/kg t.i.d. × 3 days) were unable to attenuate pituitary PRL mRNA levels. The presynaptic 5-HT autoreceptor agonist 5-MeODMT (6 mg/kg t.i.d. × 3 days) was equally ineffective. Each of these drugs, however, was able to produce marked reductions in VIP mRNA content in the anterior pituitary. This study indicates that the serotonergic system contributes to the regulation of anterior pituitary PRL and VIP mRNA expression.

Proopiomelanocortin, prolactin and growth hormone messenger ribonucleic acid levels in the fetal sheep pituitary during late gestation

We have measured the relative levels of proopiomelanocortin (POMC), prolactin (PRL) and growth hormone (GH) messenger ribonucleic acid (mRNA) in the anterior and neurointermediate lobes of the fetal pituitary during the last 2-3 weeks of gestation. The mean POMC mRNA/18S RNA ratio in the fetal anterior pituitary was significantly greater (p < 0.02) at 130-136 days (0.90 +/- 0.08; N = 9) than at 141-143 days of gestation (0.67 +/- 0.07; N = 6). In contrast, the mean PRL mRNA/18S RNA ratio increased significantly (p < 0.02) between 130 and 136 days (0.31 +/- 0.05; N = 9) when compared with 141-143 days of gestation (0.58 +/- 0.10; N = 6). There was no significant difference, however, between the mean GH mRNA/18S RNA ratio in fetal anterior pituitaries at 130-136 days (0.95 +/- 0.04; N = 9) when compared with 141-143 days of gestation (1.08 +/- 0.14; N = 6). The POMC mRNA/18S RNA ratio in the neurointermediate lobes was seven-, five- and tenfold higher than anterior pituitaries at 130-134, 135-136 and 141-143 days of gestation, respectively. We hypothesize that elevated circulating cortisol levels after 140 days of gestation act in the slow time domain (i.e. over days) to suppress POMC gene expression and that the increase in fetal pituitary PRL mRNA levels may be a consequence of oestrogen stimulation in late gestation.

A Factor(s) from a Rat Trophoblast Cell Line Inhibits Prolactin Secretion in Vitro and in Vivo1

The purpose of the present study was to measure the inhibitory action of secretions from trophoblast cells on prolactin (PRL) secretion in cycling and pregnant rats, and to determine whether factor(s) from trophoblast cells act directly on anterior pituitary cells. A rat choriocarcinoma cell line (Rcho)--a line consisting of trophoblast cells, including differentiated giant cells that secrete members of the placental PRL family--was used. When Rcho cells (1 x 10(6) cells) were transplanted under the kidney capsule of cycling rats, tumors developed and the rats went into constant diestrus. Eight days after cell injection, plasma progesterone was significantly increased in treated rats compared to controls, whereas plasma and pituitary PRL and pituitary PRL mRNA levels were significantly decreased. Similar PRL results were seen on Day 9 of pregnancy after injection of Rcho cells on Day 0 or Day 1 of pregnancy. To determine whether secretions from Rcho cells had a direct effect on anterior pituitary cells to inhibit PRL release, anterior pituitary cells were enzymatically dispersed and cultured for 4 days. Conditioned medium was obtained from 9-day Rcho cell cultures and concentrated by ultrafiltration. A fraction containing substances with molecular weights greater than 10,000 suppressed PRL release from the pituitary culture after 3 and 24 h. Conditioned medium containing substances with molecular weights between 1000 and 10,000 had no effect on PRL release, nor did conditioned medium from a placental cell line designated HRP-1. HRP-1 also contains trophoblast cells but does not contain the differentiated giant cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary luteinizing hormone and prolactin messenger ribonucleic acid levels are inversely related in laying and incubating turkey hens.

The relationships of prolactin (PRL) and LH messenger (m) RNA to serum and pituitary content were determined for turkey hens at different phases of the reproductive cycle. In the nonphotostimulated, reproductively inactive hen, serum and pituitary PRL content and pituitary PRL mRNA levels were low. All three PRL values rose after photostimulation and peaked during the incubation phase. Relative to nonphotostimulated hens, hyperprolactinemic incubating hens showed 220-, 11-, and 57-fold increases in serum PRL, pituitary PRL content, and pituitary PRL mRNA levels, respectively. These peak levels declined 80-, 3-, and 6-fold, respectively, in photorefractory hens. In contrast to PRL levels, serum LH, pituitary LH, and pituitary LH beta-subunit mRNA levels did not change as dramatically. Serum LH showed no significant changes for the different reproductive phases. Pituitary LH peaked after photostimulation and declined to its lowest level in incubating hens. Pituitary LH-beta mRNA abundance was highest in photostimulated and laying hens and lowest in incubating and photorefractory hens. These results demonstrate that the abundance of LH-beta and PRL mRNA shows an inverse relationship in photostimulated/laying and incubating turkey hens.

Depressed Prolactin Cell Activity in Long-Term Blind-Pinealectomized Female Hamsters Is Due to Loss of Estrous Cyclicity

Pinealectomy in the female golden Syrian hamster is not always completely effective in preventing the suppressive effects of long-term light deprivation due to blinding on pituitary prolactin (PRL) cell activity. We examined this curious phenomenon by measuring pituitary PRL mRNA levels, PRL synthesis, and radioimmunoassayable PRL, and correlating these changes with the status of estrous cyclicity. As expected, 12 weeks of light deprivation resulted in loss of estrous cyclicity and a greater than 90% decline in all indices of pituitary PRL cell activity, compared with intact cycling controls. Pinealectomy prevented only 40-50% of this decline. However, if noncycling light-deprived pinealectomized animals were excluded, pinealectomy was completely effective, i.e., cycling intact control animals were no different than cycling blind-pinealectomized. We conclude that the inability of pinealectomy to completely prevent the decline in prolactin cell activity seen after blinding is due to the loss of estrous cyclicity in some blind-pinealectomized females, with the attendant loss of the prolactin-stimulating hormone estrogen.

Prolactin messenger ribonucleic acid levels, prolactin synthesis, and radioimmunoassayable prolactin during the estrous cycle in the golden syrian hamster

The purpose of this study was to observe the molecular dynamics of pituitary prolactin (PRL) gene expression during the estrous cycle of the Golden Syrian hamster. PRL messenger ribonucleic acid (mRNA) levels, PRL synthesis ( 3H-PRL in the incubation media or incubated pituitary after a 3 hr incubation with 3H-leucine), and radioimmunoassayable (RIA) PRL (in the incubation media or incubated pituitary after the 3 hr incubation) were measured in the morning (0930–1100 hr) on each day of the cycle. We observed that all of these PRL indices declined or did not change from Day 2 to Day 3 of the cycle. From Day 3 to Day 4 (proestrus), however, PRL mRNA levels increased 33–38% and media 3H-PRL increased 32–42%, while there were no significant changes in pituitary 3H-PRL, or RIA-PRL in the media or pituitary. From Day 4 to Day 1 (estrus) there was a reciprocal change in the levels of 3H-PRL in the pituitary vs. the media, with the former increasing 37–50% and the latter decreasing 25–32%. Pituitary RIA-PRL also increased 45–64% from Day 4 to Day 1 while media RIA-PRL did not change. These data are consistent with the following hypothesis: On the morning of proestrus (Day 4) in the hamster, PRL mRNA levels are elevated compared to those on Day 3, signaling an increase in PRL synthesis. This newly synthesized PRL is shunted into a “readily releasable” pool on the morning of Day 4 (contributing to the afternoon surge of serum PRL), and into a “preferentially stored” pool by the morning of Day 1 (for release in response to cervical stimulation and use as a luteotrophin to maintain early pregnancy should fertilization occur).

Depressed pituitary prolactin mRNA, prolactin synthesis, and prolactin storage after light-deprivation in female hamsters is not due to loss of estrous cyclicity alone.

Pituitary prolactin (PRL) cell activity (i.e. PRL messenger ribonucleic acid [mRNA] levels, PRL synthesis, and radioimmunoassayable [RIA]-PRL), and serum RIA-PRL were measured in female golden Syrian hamsters that were (1) light-deprived and then ovariectomized before loss of estrous cyclicity, (2) light-deprived but not yet acyclic, and (3) light-deprived and ovariectomized simultaneously. The results indicate that light-deprivation can decrease PRL cell activity in ovariectomized hamsters but not in animals that continue to cycle. Thus, estrous cyclicity can be said to largely protect PRL cell activity from depressions due to light deprivation. After acyclicity/ovariectomy, however, PRL cell activity is no longer protected and light-deprivation leads to large depressions in PRL mRNA levels, PRL synthesis, and RIA-PRL beyond that caused by acyclicity/ovariectomy alone. As seen in previous studies of total light-deprivation in nonovariectomized female hamsters, we found that removing the pineal gland in conjunction with light-deprivation in ovariectomized hamsters can completely, partially, or fail to restore various measures of PRL cell activity.

The role of the suckling stimulus in regulating pituitary prolactin mRNA in the rat

Prolactin (PRL) gene expression and the synthesis and secretion of PRL were examined in ovarian-intact lactating rats suckling eight pups on 10 days postpartum. Plasma samples were assayed for PRL concentrations, and pituitary glands were analyzed for total PRL content and PRL mRNA levels. We found that suckling-induced hyperprolactinemia was associated with very high levels of plasma PRL and a doubling in pituitary PRL mRNA levels, whereas pituitary PRL content was not changed. Removal of the suckling pups decreased plasma PRL concentrations 15-fold within 24 h. This decrease in PRL secretion was not accompanied by any significant change in pituitary PRL content. Evidently, both synthesis and secretion of PRL were decreased in the pituitary gland within 24 h following cessation of suckling, as pituitary PRL mRNA content had returned to diestrous levels at this time. To determine whether or not ovarian steroids might have contributed to the changes in PRL synthesis and secretion during lactation and after withdrawal of the suckling stimulus, the experiments were repeated in lactating rats ovariectomized (OVX) on day 2 postpartum. The results in these OVX rats were qualitatively similar to those described in ovarian-intact rats. We concluded from these findings that the stimulus of suckling induces increases in PRL mRNA levels in the pituitary which provides for the increased PRL synthesis accompanying increased PRL secretion. The cessation of suckling led to prompt decreases in PRL synthesis and secretion within 24 h.

An Early Pineal-Induced Suppression of Pituitary Prolactin mRNA Levels in Light-Deprived Male Hamsters

Light deprivation by blinding or exposure to short photoperiod results in depressed prolactin (PRL) cell activity and gonadal regression in the male Syrian hamster after at least 42-56 days of treatment, and both of these events can be prevented if pineal removal accompanies light deprivation. In the present study, we investigated changes in PRL cell activity after 11, 21, or 42 days of blinding by measuring changes in: (1) pituitary PRL mRNA levels; (2) newly synthesized pituitary PRL, and (3) pituitary-RIA-PRL. The purpose of this study was to characterize the early (i.e. prior to gonadal regression and loss of reproductive function) effects of light deprivation on PRL production in the pituitary gland of the male hamster, and to determine whether a reduction in PRL mRNA levels precedes the inhibition of PRL synthesis and storage. After 11, 21, and 42 days of blinding, PRL mRNA levels in the male hamster pituitary (relative PRL mRNA/pituitary) were significantly depressed compared to intact controls by 33, 41, and 65%, respectively. Pituitary PRL synthesis (dpm of 3H-PRL/pituitary) and RIA-PRL (micrograms PRL/pituitary) were significantly depressed (by 49 and 53%, respectively) only after 42 days of blinding. All of these statistically significant depressions were prevented when the surgical blinding procedure was accompanied by pineal removal. There were no differences in testicle, pituitary, or body weight as a result of treatment. These results support the hypothesis that light deprivation by blinding causes a pineal-dependent decline in PRL mRNA levels, which leads to a decrease in PRL synthesis and pituitary RIA-PRL.