Basal Prolactin Release Research Articles

Adenosine acts by A1 receptors to stimulate release of prolactin from anterior-pituitaries in vitro.

Adenosine has been identified in the anterior pituitary gland and is secreted from cultured folliculostellate (FS) cells. To determine whether adenosine controls the secretion of anterior pituitary hormones in vitro, adenosine was incubated with anterior pituitaries. It stimulated prolactin (PRL) release at the lowest concentration used (10(-10) M); the stimulation peaked at 10(-8) M with a threefold increase in release and declined to minimal stimulation at 10(-4) and 10(-3) M. Follicle-stimulating hormone release was maximally inhibited at 10(-8) M, whereas luteinizing hormone release was not significantly inhibited. Two selective A1 adenosine receptor antagonists (10(-7) or 10(-5) M) had no effect on basal PRL release, but either antagonist completely blocked the response to the most effective concentration of adenosine (10(-8) M). In contrast, a highly specific A2 receptor antagonist (10(-7) or 10(-5) M) had no effect on basal PRL release or the stimulation of PRL release induced by adenosine (10(-8) M). We conclude that adenosine acts to stimulate PRL release in vitro by activating A1 receptors. Since the A1 receptors decrease intracellular-free calcium, this would decrease the activation of nitric oxide synthase in the FS cells, resulting in decreased release of nitric oxide (NO). NO inhibits PRL release by activating guanylate cyclase that synthesizes cGMP from GTP; cGMP concentrations increase in the lactotrophs leading to inhibition of PRL release. In the case of adenosine, NO release from the FS cells decreases, resulting in decreased concentrations of NO in the lactotrophs, consequent decreased cGMP formation, and resultant increased PRL release.

Depriving neonatal rats of milk from early lactation has long-term consequences on mammotrope development.

The sudden appearance of prolactin-releasing cells during the early postnatal period of the rat is initiated by a small milk-borne peptide. Depriving newborn rats of this early milk factor severely retards mammotrope differentiation during the neonatal period. In the present work, we extend our study of early milk deprivation to the adult. To this end, newborn litters were crossfostered onto mothers that had given birth the same day or one week earlier in order to deprive pups in the latter group of early milk. At 5, 15, and 30 d of age, rats deprived of such milk had decreased percentages of mammotropes (as measured by reverse hemolytic plaque assay, RHPA) when compared to nondeprived animals (P < 0.05). By 45 d, the percentage of mammotropes was similar for the two crossfostered groups (P > 0.1) and this persisted through d 60. Subsequently, we assessed the secretory capacity of mammotropes from 60-d old rats to secretagogues and found that early milk deprivation had no effect on basal prolactin release (P > 0.1), but that it augmented hormone secretion evoked by thyrotropin-releasing hormone (TRH, 100 nM; P < 0.01). The inhibitory response to dopamine (DA; 1 microM) and the stimulatory response to angiotensin II (AGII; 100 nM) were not altered by early milk deprivation (P > 0.1). Taken together, these results demonstrate that factors in milk from early lactation are required for normal mammotrope differentiation, and that the delay induced by early milk deprivation leads to altered secretory function of mammotropes in adult animals.

Involvement of nitric oxide in the interferon- γ-induced inhibition of growth hormone and prolactin secretion in anterior pituitary cell cultures

In previous work it was shown that the immune cytokine interferon- γ (IFN- γ) inhibits hormone secretion in anterior pituitary (AP) cell cultures, an action most likely mediated by folliculostellate (FS) cells. In the present study, we wanted to investigate whether nitric oxide (NO) is involved in this inhibitory action of IFN- γ. NO synthase (NOS) inhibitors with affinity for the inducible (iNOS) and the constitutive (cNOS) isoform such as N G-monomethyl- l-arginine ( l-NMMA) and S-methyl- l-thiocitrulline (SMLT) dose-dependently blocked the inhibitory action of IFN- γ on GHRH-stimulated GH secretion, and partially reversed the inhibitory effect on basal prolactin (PRL) release. In the absence of IFN- γ these inhibitors significantly augmented basal PRL release and slightly enhanced GHRH-stimulated GH release. l- N 6-(1-iminoethyl)lysine ( l-NIL), a NOS inhibitor with preferential affinity for iNOS, abrogated the IFN- γ effect on GHRH-stimulated GH secretion and partially reversed IFN- γ inhibition of PRL release. However, l-NIL did not exert a stimulatory effect on basal PRL and GHRH-stimulated GH release by its own. 2,4-diamino-6-hydroxypyrimidine (DAHP), a NOS inhibitor by interfering with tetrahydrobiopterin (BH4) cofactor availability, showed the same activity profile as l-NIL. NOS inhibitors blocked or reduced the production of NO as detected by measuring nitrite (NO − 2) levels in AP cell cultures and cGMP levels in the NO-reporter cell line RFL-6. The NOS inhibiting action of l-NMMA was confirmed by competition experiments with the natural NOS substrate l-arginine. Thus, in culture medium with lower amounts of l-arginine, l-NMMA blocked the IFN- γ-induced inhibition of GHRH-stimulated GH release at a lower dose. The inhibition of PRL and GH release by IFN- γ was markedly reduced in l-arginine-depleted medium. The NO donor sodium nitroprusside (SNP) mimicked the inhibitory action of IFN- γ on GHRH-stimulated GH and basal PRL release. Similarly to IFN- γ, SNP did not affect basal GH release. As previously reported, inhibition by IFN- γ occurred only in AP cell populations containing a minimal proportion of FS cells. As studied in different cell populations obtained by unit gravity sedimentation in a serum albumin gradient, l-NMMA reversed the IFN- γ effect in the same populations enriched in FS cells. Interestingly, in the absence of IFN- γ l-NMMA strongly stimulated basal PRL release in the population most enriched in FS cells. It is concluded that IFN- γ through activation of the iNOS pathway probably in FS cells enhances the production of NO and that this effect is responsible for the inhibitory action of IFN- γ on GHRH-stimulated GH release and partially for the IFN- γ-induced decrease in basal PRL release. On the other hand, NO, likely produced by cNOS, appears to exert a tonic inhibitory effect on GHRH-stimulated GH and basal PRL release. It seems therefore that low amounts of NO produced constitutively may take charge of subtle physiological adaptations, and higher levels of NO produced by iNOS under the influence of IFN- γ may attenuate PRL and GH release during emergency conditions of immune and inflammatory reactions.

Anterior pituitary release of prolactin is inhibited by exposure to short photoperiod.

The potential regulatory sites responsible for the decrease of circulating prolactin (PRL) levels shown by many photoperiodic mammals following prolonged exposure to short days was investigated in Siberian hamsters that had been maintained under a stimulatory, long-day photoperiod, and in hamsters that had been shifted to a nonstimulatory, short-day photoperiod for 8-10 weeks. The ability of anterior pituitary fragments (AP) from each of these groups to release prolactin was evaluated in pituitary tissue cultured alone and also in pituitary tissue co-cultured with hypothalamic fragments (HF), using a perifusion tissue culture system. The perfusate from these cultures was collected every 1/2 h for 8 h, and was assayed for basal levels of prolactin using radioimmunoassay. For AP tissue cultured alone, there was a robust reduction in prolactin release by the fragments harvested from short-day housed animals. In AP tissue harvested from long-day exposed animals, co-culture with either long- or short-day HF did not induce significant changes in basal PRL release. Similarly, co-culture with short-day HF did not significantly alter PRL release in short-day APs. However, there was a significant increase in release when short-day APs were co-cultured with long-day HF. These results suggest a direct effect of photoperiod on PRL synthesis and/or release at the level of the pituitary. However, the altered responsiveness of short-day pituitaries could be the result of previous, chronic inhibitory hypothalamic input during short-day exposure. A follow-up study was conducted to investigate the ability of vasoactive intestinal peptide (VIP) to stimulate PRL release from long- and short-day APs. Results indicated that the ability of VIP to stimulate PRL release is both photoperiod and dose dependent.

Transient effects of MK-801 administration on secretion of luteinizing hormone and prolactin in ovariectomized and estradiol-treated sheep.

The neuroendocrine role of endogenous ligands for the excitatory amino acid receptor subtype known as the NMDA receptor was investigated by administering the NMDA receptor antagonist MK-801 to ovariectomized (ovx) and estradiol-treated sheep. Repetitive administration of MK-801 at intravenous (iv) dosages of 0.1 mg/kg to untreated ovx ewes did not affect the episodic profiles of luteinizing hormone (LH) release, but each injection of MK-801 abruptly stimulated release of prolactin (PRL) demonstrating the effectiveness of the dosage. Injection of estradiol-17beta (50 microg/ewe) to ovx ewes produced the expected biphasic LH response; an initial suppression followed by a surge-like LH increase together with an elevated basal secretion of PRL. Injections of MK-801 occurring 9 and 11 h after estradiol-17beta injection rapidly and transiently increased serum LH in a very unexpected response. However, these same MK-801 injections also resulted in decreased serum LH 14-17 h after estradiol-17beta by delaying the onset of the surge-like release of LH. Estradiol-17beta administration itself elevated basal release of PRL to serum concentrations observed after repetitive MK-801, and further injection of MK-801 no longer transiently increased serum PRL as it had done prior to estradiol-17beta treatment. In summary, antagonizing the endogenous excitatory amino acid ligands of the NMDA receptor with MK-801 did not alter either the timing or magnitude of the putative episodic discharges of gonadotropin-releasing hormone (GnRH) which in turn cause the episodic releases of pituitary LH in ovx sheep. Estrogen administration created a transient neuroendocrine environment in which antagonism of these endogenous ligands was stimulatory to abrupt discharge of GnRH and thereby to acute release of LH. Antagonism of NMDA receptors in untreated ovx ewes stimulated release of PRL suggesting that the endogenous NMDA ligands were probably stimulatory to the release of a PRL-inhibiting neurohormone such as dopamine.

Age-dependent prolactin-releasing activity of nucleoproteins

In previous studies we demonstrated that histone preparations possess multiple effects in vivo on pituitary hormone secretion and that these effects tend to disappear with age. We have now evaluated the in vitro effects of histone and nucleohistone preparations on the secretion of prolactin (PRL) in perifused pituitary cells from young (4 months) and senescent (29 33 months) female rats. Freshly dispersed pituitary cells were packed into short columns and were continuously perifused with serum-free medium. The substances to be tested were pumped through the perifusion circuit, at the end of which perifusate fractions were collected and hormones measured by specific radioimmunoassay (RIA). Quantitative immunohistochemistry was carried out on the pituitary glands from seven young and six senescent females. In vitro basal PRL release was similar in both age groups. Perifusion of cells with median eminence extract ( 1 90 to 1 10 ), histone H2A (100 to 1000 μg/ml) or nucleohistone (200 to 1000 μg/ml), generated PRL responses which were higher in young than in senescent cells. The pituitaries of the senescent animals were characterized, in most cases, by the presence of chromophobic microprolactinomas against a background of diffuse prolactotroph hyperplasia. Our results confirm previous evidence that circulating nucleohistones and histones may act as hypophysotropic signals. The morphologic alterations in PRL cell populations found in the senescent rats may play a role in the desensitization of the pituitary gland to nucleoproteins, and possibly to other hypophysiotropic molecules, with age.

Calcitonin-like peptide containing gonadotrophs are juxtaposed to cup-shaped lactotrophs

Calcitonin (CT) is known to inhibit basal and TRH-stimulated prolactin release in cultured anterior pituitary cells in vitro and pituitary CT-like peptide (pit-CT) is synthesized and released by isolated anterior pituitary cells. However, the specific cell type containing pit-CT has not been identified. To determine this, double label immunohistochemistry was performed on pituitary sections from male rats using antisera for specific marker peptides of gonadotrophs, thyrotrophs, lactotrophs, somatotrophs, corticotrophs, and folliculo-stellate cells. CT was only colocalized with gonadotroph-specific markers and the distribution of pit-CT immunoreactive (IR) cells followed the patterns of gonadotroph distribution in male and female rats. Double and triple label immunohistochemistry using antiserum for CT, FSH, and PRL showed an apposition of calcitonin-like peptide containing gonadotrophs to cup-shaped lactotrophs. To examine whether pit-CT IR was altered, similarly to gonadotrophs, with known changes in PRL serum levels, studies were extended to ovariectomized, pregnant, and lactating rats. The area covered by pit-CT immunoreactivity and the tissue content of pit-CT significantly differed between physiological states and the pit-CT level was inversely related to the known PRL status. Pit-CT containing gonadotrophs were in all cases apposed to cup-shaped lactotrophs. These results provide histological support for previous studies proposing that pit-CT serves as a paracrine inhibitor of PRL release.

Role of the Na +-Ca 2+ and Na +-H + antiporters in prolactin release from anterior pituitary cells in primary culture

2′,4′-dimethylbenzamilamiloride (DMB), a somewhat selective inhibitor of the Na +-Ca 2+ exchanger, in concentrations of 10, 30 and 100 μM did not produce any significant effect on baseline prolactin release from anterior pituitary cells in primary culture. When prolactin secretion was stimulated by the inhibitor of the Na +-K +-ATPase, ouabain, that activates the Na +-Ca 2+ exchanger as a Ca 2+-influx pathway, DMB was able to produce inhibition of prolactin secretion. 5-( N,N-hexamethylene) amiloride (HMA), another amiloride analog which specifically inhibits the Na +-H + antiporter and has no inhibitory activity on the Na +-Ca 2+ exchanger, at the concentrations of 0.1, 1 and 10 μM, did not affect basal prolactin release whereas it significantly reduced prolactin release stimulated by thyrotropin releasing hormone (TRH) (1 μM). These results suggest that the Na +-Ca 2+ antiporter is involved in the process of prolactin release elicited by the inhibition of the Na +-K +-ATPase whereas the Na +-H + antiporter is involved in the prolactin secretion elicited by TRH.

Permissive role of corticotropin-releasing factor in the acute stress-induced prolactin release in female rats

The present study examined the role of corticotropin-releasing factor (CRF) in the acute stress-induced release of prolactin (PRL) in ovariectomized estrogen-primed female rats. Acute immobilization stress induced a marked increase in serum PRL levels in animals treated with saline intraventricularly (icv). However, a prior icv injection of a-helical CRF(9–41), a CRF antagonist, completely eliminated the immobilization-induced PRL release in the majority of animals, providing evidence for involvement of CRF in the acute stress-induced PRL release. On the other hand, an icv injection of CRF did not affect basal PRL release at any dose in non-stressed animals, suggesting that the peptide plays a permissive role which enables other undefined stress mediator(s) to stimulate PRL release.

Estrogen attenuates expression of calcitonin-like immunoreactivity in the anterior pituitary gland

Estrogens increase prolactin (PRL) synthesis and release in rats and humans, whereas pituitary-derived calcitonin-like immunoreactive peptide (pit-CT) inhibits PRL gene expression and release. To test the hypothesis that estrogens stimulate lactotrophs by diminishing pit-CT expression, the present studies examined effects of ovariectomy (ovx) and estradiol (E2) administration on (1) pit-CT IR cell population; (2) pit-CT IR content and (3) release of pit-CT IR by cultured anterior pituitary (AP) cells. Ability of anti-calcitonin immunoglobulins (anti-CT IgG) to stimulate PRL release from cultured AP cells was also examined. The results suggest that ovariectomy induced a large increase in pit-CT IR cell population in the AP gland and E2-treatment dramatically reversed this increase. Similar changes were observed in pit-CT IR content of AP extracts. Cultured AP cells from ovx rats released significantly higher amounts of pit-CT IR, and anti-CT IgG induced a significant increase in basal PRL release. AP cells from E2-treated rats secreted lower amounts of pit-CT IR and this was associated with significantly higher PRL release. These results suggest that estrogens may stimulate lactotrophs, at least in part, by removing inhibitory influence of endogenous pit-CT.

Neuropeptide-E-I antagonizes the action of melanin-concentrating hormone on stress-induced release of adrenocorticotropin in the rat.

The physiological role of melanin-concentrating hormone (MCH) in mammals is still very elusive, but this peptide might participate in the central control of the hypothalamopituitary adrenal (HPA) axis during adaptation to stress. Cloning and sequencing of the rat MCH (rMCH) cDNA revealed the existence of additional peptides encoded into the MCH precursor. Among these peptides, neuropeptide (N) glutamic acid (E) isoleucine (I) amide (NEI) is co-processed and secreted with MCH in rat hypothalamus. In the present work we examined: (1) The pattern of rMCH mRNA expression during the light and dark conditions in the rat hypothalamus and (2) The effect of intracerebroventricular (ICV) injections of rMCH and NEI in the control of basal or ether stress-modified release of corticotropin (ACTH), prolactin (PRL) and growth hormone (GH) secretion in vivo in light-on and light-off conditions. Our data indicate that rMCH mRNA levels do not change during the light-on period, but increase after the onset of darkness. Either alone or co-administered, rMCH and NEI do not modify basal secretion of GH and PRL at any time tested nor do they alter ether stress-induced changes in these two hormonal secretions. At the end of the light on period corresponding to the peak of the circadian rhythm in ACTH, administration of rMCH but not NEI leads to a decrease in ACTH levels while MCH is not effective during the light off period of the cycle (i.e. when basal ACTH levels are already low). Using a moderate ether induced stress, ACTH levels are only stimulated during the dark phase of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

Prolonged effects of tumor necrosis factor-alpha on anterior pituitary hormone release.

We examined the chronic (72 h) effects of 30 ng/ml recombinant murine tumor necrosis factor (TNF)-alpha on release of immunoreactive growth hormone (GH), prolactin (PRL), thyrotropin (TSH), and TSH glycosylation, as assessed by lectin binding, in cultured rat anterior pituitary cells. In cultured cells from adult female rats, TNF-alpha significantly suppressed basal and GH-releasing hormone (GRH)-stimulated GH release. TNF-alpha also suppressed basal PRL release and completely abolished the PRL response to TRH (0.1-10 nM). Whereas TNF-alpha reduced basal TSH release, it significantly enhanced the maximal TSH response to TRH. TNF-alpha did not affect the concanavalin A and lentil lectin binding of TSH accumulated in the medium during the 4-day culture, but significantly decreased the lentil lectin binding of TSH released in response to acute TRH stimulation. TNF-alpha significantly enhanced the inhibitory effect of somatostatin on stimulated PRL release, but not on GH or TSH release. Compared to cell cultures from adult female rats, in anterior pituitary cell cultures from 12-day-old rats the effects of prolonged exposure to TNF-alpha on hormone release were diminished or absent. Pituitary hormone release was unaffected by acute (3 h) exposure to TNF-alpha. These results demonstrate a direct effect of TNF-alpha on anterior pituitary hormone release, which is cell-type specific and age dependent.

Endothelin-1 modulates renin and prolactin release from human decidua by different mechanisms

Endothelin (ET)-1 stimulates the synthesis and release of renin and inhibits the expression of prolactin (PRL) from term human decidual cells. To examine the mechanisms by which ET-1 exerts its differential effects on renin and PRL expression, we have studied total renin and PRL release from term human decidual cells in response to pharmacological agents that affect calcium- and protein kinase C-dependent mechanisms. Calcium ionophore A-23187 stimulated basal renin release and potentiated ET-1-stimulated renin release but had no effect on basal or ET-inhibited PRL release. The calcium channel blocker nifedipine inhibited ET-1-stimulated renin release but had no effect on PRL release. The protein kinase C agonist phorbol 12-myristate 13-acetate (PMA) stimulated basal renin release and potentiated the effect of ET-1 on renin release. However, PMA inhibited basal PRL release and also enhanced the inhibitory effect of ET-1. The PKC inhibitor staurosporine increased basal PRL release and completely reversed the inhibitory effect of ET on PRL release. These results indicate that the effects of ET-1 on both decidual renin and PRL release are dependent on the activation of protein kinase C. However, the effect of ET-1 on renin release appears to be dependent on extracellular calcium, whereas the effect on PRL is not influenced by extracellular calcium.

Alteration of basal release of anterior pituitary hormones by pretreatment of primary cultured cells with trypsin

The anterior pituitary (AP) gland secretes 6 different hormones. Prolactin (PRL) is secreted at a relatively high level without stimulation by the hypothalamus, while secretion of the others requires the action of stimulatory factors from the hypothalamus. In order to gain an insight into the mechanism underlying the different spontaneous release patterns of these hormones, we investigated their spontaneous release rate after pretreating rat anterior pituitary cells with trypsin. Rat AP cells were cultured on Cytodex microcarrier beads for 4 days and were then superfused with either control medium or medium containing trypsin (0.25%) for 5 min. The subsequent release rates of the AP hormones were monitored. The basal release of PRL was severely reduced to almost undetectable level and began to recover 120 min after the trypsin-pretreatment. Full recovery was attained over the next 100 min and was delayed by treatment with a protein synthesis inhibitor, cycloheximide (7 μM). In the trypsin-pretreated cells, basal release of PRL and growth hormone (GH) was severely reduced, while that of thyroid stimulating hormone (TSH) and adrenocorticotropic hormone (ACTH) was enhanced and luteinizing hormone (LH) and follicle stimulating hormone (FSH) was not markedly affected by the treatment, suggesting that the suppression of PRL release was not caused by nonspecific damage to the cells. Since trypsin does not readily enter cells, the altered secretion of AP hormones seems to be the result of restricted digestion of the external components of the cells. On the bases of these observations, we predicted that the mechanism of spontaneous release of hormones involves trypsin sensitive proteins (TSMP) on the plasma membranes of the anterior pituitary cells.

Vasoactive Intestinal Polypeptide Antiserum Affects Rat Prolactin mRNA in 40-Day but Not 110-Day Diethylstilbestrol-Induced Prolactinoma Tissue

Experiments were designed to examine whether vasoactive intestinal polypeptide (VIP), a known stimulator of basal prolactin (PRL) secretion, regulates PRL gene expression in the rat diethylstilbestrol (DES)-induced prolactinoma model. The VIP-induced increase in PRL release could result from increased PRL synthesis and/or decreased PRL degradation. Male Fischer 344 rats were implanted with 10 mg DES pellets 40 or 110 days prior to obtaining the anterior pituitary glands for cell dispersal. Cells were incubated in 1:10 normal rabbit serum or VIP antiserum (AVIP). After incubation, cells were pelleted, washed, and pooled for total nucleic acid extraction. The rat PRL (rPRL) mRNA abundance was quantitated using a solution hybridization/ribonuclease protection assay. Supernatant was collected and analyzed for PRL content using radioimmunoassay. Results from this experiment reveal partial immunoneutralization of intrapituitary VIP significantly decreased PRL secretory rate by rapid reduction in rPRL mRNA in the 40-day tumors. However, in the 110-day tumors the rPRL mRNA steady-state levels were unchanged but the basal release of PRL continued to be decreased by AVIP. These results indicate VIP exerts its effects on PRL secretion through at least two mechanisms.

Effects of LHRH and ANG II on prolactin stimulation are mediated by hypophysial AT1 receptor subtype.

We have used the nonpeptide angiotensin II (ANG II) receptor antagonists losartan (receptor subtype AT1) and PD-123319 (AT2) to determine the participation of ANG II receptor subtypes in luteinizing hormone-releasing hormone (LHRH)-induced prolactin release in a perifusion study using intact pituitaries in vitro. LHRH (1.85 x 10(-7) M) released prolactin consistently, whereas losartan (10(-5) M) abolished prolactin response without modifying basal prolactin or luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release. PD-123319 (10(-5) M) had no effect on basal or LHRH-induced prolactin, LH, or FSH release. We also determined that the effect of ANG II on prolactin release was mediated by the same receptor subtype. In adenohypophysial cells dispersed in vitro ANG II (10(-8) M) released prolactin. Losartan (10(-7) and 10(-6) M), but not PD-123319, inhibited this effect. We conclude that in intact hypophyses of 15-day-old female rats the effect of LHRH on prolactin release is readily demonstrated. LHRH-induced prolactin release appears to be mediated by ANG II acting in a paracrine manner on AT1 receptors located on lactotrophs.

Role of laminin on prolactin and gonadotrophin release from anterior pituitaries of male rats

The effect of rat laminin on the release of prolactin (PRL), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from anterior pituitaries of adult male rats was studied in vitro. Laminin had a bidirectional effect on basal release of PRL, being stimulatory at 10 −12M and inhibitory at 10 −9 and 10 −8M. LH release was only stimulated by the 10 −12M concentration and no changes in FSH release were observed. Laminin did not modify the stimulatory action of thyrotropin-releasing hormone (TRH) nor luteinizing hormone releasing hormone (LHRH), but the response of FSH to LHRH was inhibited with a maximal effect at 10 −10M. To assess its physiological role, the effect of a rabbit antibody to laminin on the release of PRL, LH and FSH was also studied. IgG anti-laminin on at 2 × 10 −7M had highly significant stimulatory effect on PRL release after 3 and 5 hr of incubation, as compared to IgG from normal rabbit serum and medium alone used as controls. No changes on the release of LH and FSH were detected. These results suggest that laminin plays a physiologically significant inhibitory role on PRL release at the pituitary level in vitro.

Effects of age and sex on basal and dopamine-inhibited in vitro prolactin release, and dopamine receptor binding, by the rat adenohypophysis

We measured basal and dopamine-inhibited pituitary cell prolactin (PRL) release in vitro, and dopamine receptor binding in pituitary homogenates, from intact male and female Wistar rats of varying ages. During 48-72 hours in culture, the baseline secretion rate of PRL from pituitary cells of old (24 months) male rats was less than one-half that from cells of mature (6 months) male rats, whereas the corresponding basal secretion rate of PRL from cells of old female rats was nearly 3-fold greater than that from cells of mature female rats (p < 0.001). After in vitro exposure to various concentrations of dopamine (10(-10)M to 10(-6)M), PRL secretion decreased from pituitary cells of both mature and old rats (p < 0.001). However, repeated measures analysis of variance revealed an age-related dose-dependent decrease in the magnitude of dopamine-inhibited PRL release from cells of both male and female rats (p < 0.001). Dopamine receptor number did not differ with age (3-25 months), but was 2 to 3-fold greater in female than in male rats (p < 0.01). Receptor affinity was decreased with age only in female rats, and was greater in female than in male rats (p < 0.05). These data suggest that the decrease in dopamine-inhibited PRL release from pituitary cells of old male and female rats is not due to altered pituitary dopamine receptor binding, despite certain sex differences.

Galanin regulates basal and oestrogen-stimulated lactotroph function.

Oestrogen, an essential physiological regulator of reproductive function, controls lactotroph proliferation and prolactin release. The neuropeptide galanin co-localizes to the lactotroph, but its physiological function is largely unknown. Pituitary galanin expression is extremely sensitive to the oestrogen status of the animal. A marked elevation occurs during pregnancy and lactation, and exogenous 17 beta-oestradiol can cause a 4,000-fold increase in messenger RNA levels. Here we report that galanin is secreted by a minority of lactotrophs and is essential for the regulation of basal and vasoactive-intestinal-polypeptide-stimulated prolactin release. Hyperoestrogenization increases the number of galanin-secreting cells and the resulting increase in basal prolactin release is completely abolished by treatment with galanin antiserum. Galanin is a potent lactotroph growth factor and galanin-immunoneutralization completely inhibits the previously reported mitogenic effects of oestrogen on the lactotroph. These findings represent direct evidence for paracrine regulation of lactotroph function and demonstrate that the effect of oestrogen on lactotroph proliferation and prolactin release are mediated by locally secreted galanin.

MMQ cells: a model for evaluating the role of g proteins in the modulation of prolactin release

It is well known that dopamine (DA) inhibits while vasoactive intestinal peptide (VIP) and angiotensin II (ANG II) stimulate prolactin (PRL) release from normal anterior pituitary lactotrophs; however, elucidation of the intracellular mechanisms involved in these effects has been hindered by the cellular heterogeneity of the anterior pituitary. MMQ cells, isolated from the PRL-secreting rat pituitary tumor 7315a is an interesting model since they only secrete PRL. In order to determine whether and which GTP-binding (G) proteins are involved in the modulation of cyclic 3',5'-adenosine monophosphate (cAMP) accumulation and phospholipids turnover and eventually PRL release, we have performed studies with MMQ cells. For this purpose, the levels of various G proteins (αo, αs, αi, αq and β) and their mRNAs, measured by Western and Northern blots respectively, were correlated with intracellular cAMP accumulation in response to DA, VIP or DA plus VIP, and with inositol phosphates (IPx) formation in response to ANG II, DA or DA plus ANG II. This study shows that, when compared to normal pituitary tissue, the levels of αo, αo2 and αi3 were significantly decreased in MMQ cells; those of αol, αi (αil + αi2), αs42 and αq were very low or undetectable while those of αs47 and β were normal. DA was unable to inhibit basal PRL release and cAMP accumulation. VIP increased both cAMP accumulation and PRL release, while cAMP accumulation elicited by VIP could be suppressed by DA. BAY K 8644-induced PRL release also could be suppressed by DA. DA and, paradoxically, ANG II decreased IPx formation; the action of ANG II could be blocked by SAR,ILE-ANG II and was exerted through pertussis toxin-sensitive G proteins. This study and previously published results provide circumstantial evidence that αo, αil and αi2 are all involved in the transduction of the DA inhibitory message while αs47 transduces cAMP activating messages and αs42 is responsible for the constitutive activation of L-type Ca 2+ channels, adenylate cyclase and baseline PRL release.