Interactions between prolactin and kisspeptin to control reproduction

ABSTRACT Prolactin is best known for its effects of stimulating mammary gland development and lactogenesis. However, prolactin is a pleiotropic hormone that is able to affect several physiological functions, including fertility. Prolactin receptors (PRLRs) are widely expressed in several tissues, including several brain regions and reproductive tract organs. Upon activation, PRLRs may exert prolactin’s functions through several signaling pathways, although the recruitment of the signal transducer and activator of transcription 5 causes most of the known effects of prolactin. Pathological hyperprolactinemia is mainly due to the presence of a prolactinoma or pharmacological effects induced by drugs that interact with the dopamine system. Notably, hyperprolactinemia is a frequent cause of reproductive dysfunction and may lead to infertility in males and females. Recently, several studies have indicated that prolactin may modulate the reproductive axis by acting on specific populations of hypothalamic neurons that express the Kiss1 gene. The Kiss1 gene encodes neuropeptides known as kisspeptins, which are powerful activators of gonadotropin-releasing hormone neurons. In the present review, we will summarize the current knowledge about prolactin’s actions on reproduction. Among other aspects, we will discuss whether the interaction between prolactin and the Kiss1-expressing neurons can affect reproduction and how kisspeptins may become a novel therapeutic approach to treat prolactin-induced infertility.

Effects of melatonin and prolactin in reproduction: review of literature / Efeitos da melatonina e da prolactina na reprodução: revisão de literatura.

Summary The pineal gland is responsible for producing a hormone called melatonin (MEL), and is accepted as the gland that regulates reproduction in mammals. Prolactin (PRL) also exhibits reproductive activity in animals in response to photoperiod. It is known that the concentrations of PRL are high in the summer and reduced during winter, the opposite of what is seen with melatonin in these seasons. In placental mammals, both prolactin and melatonin affect implantation, which is considered a critical point of pregnancy, since a successful pregnancy requires the development of a synchronous interaction between the endometrium and blastocyst for placental development. It is also known that PRL levels during pregnancy are essential for the maintenance of pregnancy, because this hormone induces the corpus luteum to produce progesterone, in addition to stimulating blastocyst implantation to maintain pregnancy and form the placenta. However, melatonin levels in plasma have also been shown to increase during pregnancy, peaking at the end of this period, which suggests that this hormone plays an important role in the maintenance of pregnancy. Thus, it is clear that treatment with prolactin or melatonin interferes with the processes responsible for the development and maintenance of pregnancy.

Resumo A glândula pineal é responsável pela produção do hormônio melatonina (MEL), sendo aceita como a glândula reguladora da reprodução em mamíferos. A prolactina (PRL) também exibe atividade reprodutiva em animais, em resposta ao fotoperíodo. Sabe-se que as concentrações de PRL são elevadas durante o verão e baixam durante o inverno, ocorrendo o oposto com os níveis do hormônio melatonina nessas estações. Nos mamíferos placentários, tanto a melatonina quanto a prolactina influenciam a implantação, que é considerada o ponto crítico da gravidez, pois o sucesso da gestação requer o desenvolvimento de uma interação sincronizada entre o endométrio e o blastocisto para o desenvolvimento da placenta. Sabe- -se ainda que os níveis de PRL durante a gestação são essenciais para a manutenção da gravidez, pois esse hormônio induz o corpo lúteo a produzir progesterona, além de estimular a implantação do blastocisto, mantendo a prenhez e o desenvolvimento placentário. Em contrapartida, tem-se demonstrado também que os níveis de melatonina no plasma aumentam durante a gestação, atingindo valores elevados no fim desse período, sugerindo que esse hormônio desempenhe um importante papel na manutenção da gestação. Dessa forma, fica claro que o tratamento com prolactina ou melatonina interfere nos processos responsáveis pelo desenvolvimento e pela manutenção da gestação.

Increased expression of SNARE proteins and synaptotagmin IV in islets from pregnant rats and in vitro prolactin-treated neonatal islets

During pregnancy and the perinatal period of life, prolactin (PRL) and other lactogenic substances induce adaptation and maturation of the stimulus-secretion coupling system in pancreatic â-cells. Since the SNARE molecules, SNAP-25, syntaxin 1, VAMP-2, and synaptotagmins participate in insulin secretion, we investigated whether the improved secretory response to glucose during these periods involves alteration in the expression of these proteins. mRNA was extracted from neonatal rat islets cultured for 5 days in the presence of PRL and from pregnant rats (17th-18th days of pregnancy) and reverse transcribed. The expression of genes was analyzed by semi-quantitative RT-PCR assay. The expression of proteins was analyzed by Western blotting and confocal microscopy. Transcription and expression of all SNARE genes and proteins were increased in islets from pregnant and PRL-treated neonatal rats when compared with controls. The only exception was VAMP-2 production in islets from pregnant rats. Increased mRNA and protein expression of synaptotagmin IV, but not the isoform I, also was observed in islets from pregnant and PRL-treated rats. This effect was not inhibited by wortmannin or PD098059, inhibitors of the PI3-kinase and MAPK pathways, respectively. As revealed by confocal laser microscopy, both syntaxin 1A and synaptotagmin IV were immunolocated in islet cells, including the insulin-containing cells. These results indicate that PRL modulates the final steps of insulin secretion by increasing the expression of proteins involved in membrane fusion.

Sitios de acción inhibitoria de la prolactina sobre la síntesis de estradiol inducida por la hormona folículo estimulante en las células de la granulosa / Sites of prolactin inhibition on gonadotropin-induced estrogen production in cultured rat granulosa cells

En este estudio se investigaron los sitios probables de la acción inhibitoria de prolactina (Prl) sobre la esteroidogénesis ovárica inducida por la hormona folículo estimulante (FSH). Para esta finalidad se estudió la capacidad de cultivos primarios de células de la granulosa de la rata de sintetizar estradiol y AMPc bajo la estimulación con FSH o de activadores de la vía dependiente de AMPc en presencia de Prl humana. La participación de otros sistemas de transducción de señal como los dependientes de PKC y proteínas Gi en los mecanismos de acción inhibitoria de la Prl fue también investigada utilizando inhibidores de estos sistemas como la calfostina C y la toxina pertusis. Los resultados demostraron la habilidad de la Prl de alterar la esteroidogénesis previa y posterior a la generación de AMPc, muy probablemente por mecanismos que involucran la activación de la subunidad catalítica de la adenilato ciclasa, así como a través de interactuar con sistemas de transducción de señal dependientes de PKC y proteínas sensibles a la toxina pertusis. Nuestros resultados sugieren un mecanismo de interacción entre receptores acoplados a proteínas G con aquéllos acoplados a cinasas de tirosinas mediado muy probablemente por vías de señalización dependientes de proteínas Gi.

We studied the sites of prolactin inhibition upon FSH induced ovarian steroidogenesis and the ability of prolactin (Prl) to inhibit the synthesis of estradiol and cAMP accumulation under the stimulation of FSH or cAMP dependent activators. The participation of other signal pathways such as PKC and Gi proteins on the inhibitory actions of Prl was also investigated using calfostine C andpertusis toxin as inhibitors. Results showed a dose dependent prolactin decrease in FSH-induced estradiol and cAMP production prior and after the generation of the cyclic nucleotide by a mechanism involving the catalytic subunit of adenyl cyclase and/or through activation of PKC or by the interaction with pertusin toxin sensitive G proteins. Our results suggest a mechanism by which G protein coupled receptors are linked with those coupled with tyrosine kinase through the involvement of a Gi protein mediated mechanism.

Effects of ovine LH, GH and prolactin, and testosterone on serum testosterone and estradiol-17 beta levels, and seminal vesicle and testicular activity in the catfish Clarias batrachus (L.).

Intraperitoneal administrations of testosterone (0.5 microgram/g body wt), and ovine LH (1.0 microgram/g body wt), GH (5 micrograms/g body wt) and prolactin (10 micrograms/g body wt) daily for 7 days during early prespawning phase (May) in C. batrachus produced varied effects on seminal vesicle (SVSI) and testicular (GSI) weights and biochemical correlates. Testosterone and LH treatments significantly increased serum testosterone level and concentrations of total proteins, fructose, hexosamines and sialic acid in both seminal vesicles and testis. Serum E2 levels increased significantly only after testosterone treatment. GH treatment increased significantly serum testosterone level and only the concentrations of SV hexosamines and testicular protein. Prolactin, however, significantly lowered serum testosterone level and concentrations of total protein, hexosamines in both SV and testis, and testicular fructose and sialic acid levels. The results show that the stimulating effect of LH and GH on SV and testicular activity is mediated through the increased secretion of testosterone and the inhibitory effect of prolactin by decreased testosterone secretion.

Characterization of anionic amino acid transport systems in mouse mammary gland.

L-glutamate was transported into mammary tissue via Na(+)-dependent system XAG- that strongly interacted with both D- and L-isomers of aspartate but only with L-isomer of glutamate. Replacement of Cl- by gluconate from the extracellular medium did not affect the uptake of L-glutamate. Although neutral amino acids weakly inhibited the uptake of L-glutamate, there was no evidence for the heterogeneity of anionic amino acid transport system. The XAG- system was inhibited by sulfhydryl group blocking reagent N-ethylmalemide. Low pH (6) partially inhibited the uptake by L-glutamate by mammary tissue. Prior loading of mammary tissue with L-glutamate slightly down regulated its uptake. Culturing pregnant mouse mammary tissue explants in vitro in the presence of lactogenic hormones (insulin plus cortisol plus prolactin) did not affect appreciably the uptake of L-glutamate.

Effects of gonadotropins and prolactin on ovarian activity of a wild avian species, the tree pie Dendrocitta vagabunda.

Ovine FSH (40: g per bird daily for 10 days) increased ovarian weight, follicular size, phosphatase activities, and RNA and protein levels in tree pie (Dendrocitta vagabunda), but exogenous ovine LH (40 micrograms per bird daily for 10 days) with the same dose and duration caused depletion of ovarian cholesterol and ascorbic acid concentrations with a rise in sialic acid and glycogen levels of the ovary. In contrast, prolactin (LTH: 5 I.U. per bird daily for 10 days) administration showed reverse biochemical changes to those of FSH. The findings suggest that FSH induces mainly ovarian follicular growth and LH stimulates ovarian steroidogenesis, but LTH is antigonadal in this wild avian species.

Prolactina como inmunomodulador / Prolactin how immunomodulator

El propósito de la presente revisión es explorar el papel de la prolactina como inmunomodulador en la respuesta inmune. La prolactina tiene función trófica en la proliferación de los linfocitos. Las células del sistema inmunitario tienen receptores en su superficie para la prolactina, más aún, los linfocitos son capaces de sintetizar y secretar prolactina. Diferentes estados en el nivel de prolactina ejercen una respuesta diferente en el sistema inmunitario, la disminución en la prolactina provoca un deterioro en la respuesta inmunitaria, mientras que el aumento de la prolactina ejerce un incremento de la respuesta inmunitaria. Las alteraciones en la prolactina se han descrito en muchas enfermedades con fondo inmunológico, como el lupus eritematoso sistémico, el síndrome de Reiter, artritis por adyuvantes, uveítis, transplante de órganos. La acumulación de pruebas al momento actual del papel que juega la prolactina como inmunomodulador puede tener un profundo impacto clínico en las enfermedades autoinmunitarias pero aún están en camino de determinarse

Rejuvenation of epididymal principal cells by prolactin under androgen deficiency.

Influence of prolactin on the ultrastructure of principal cells lining the epididymal epithelium was investigated in Wistar rats. Orchidectomy produced degenerative changes suggesting that structural integrity of principal cell is maintained by factors originating in the testis. The atrophic changes in the principal cell of ordhidectomised rats were significantly reversed when prolactin was administered to these animals. The number of cells that responded were found to increase with the dose of prolactin injected. On the otherhand, bromocryptine treatment did not appreciably change the ultrastructure of principal cells in orchidectomised rats. Results suggest that prolactin may have a rejuvenating epididymal principal cells in androgen deficient states.

Impact of prolactin on epididymal lipid profile in castrated rats.

Prolactin treatment to castrated rats led to accumulation of triacylglycerol and esterified cholesterol. There was no appreciable drift in epididymal cholesterol: phospholipid ratio between the prolactin treated and control animals. However, further analysis of phospholipids showed a build up of phosphatidyl inositol, phosphatidyl choline and phosphatidyl ethanolamine but a drop in the levels of phosphatidyl serine and sphingomyelin in prolactin treated castrated rats as compared to those castrated animals injected with vehicle alone. Changes in phospholipids reported above were prominently seen in the group of castrated rats that received 100 micrograms oPRL/100 g body weight but not in those animals which received either lower or higher doses of the hormone. Interestingly, bromocryptine treatment in castrated rats produced a general depletion in the levels of all lipid classes studied in the epididymis. It is suggested that this may be due to impaired synthesis and/or increased breakdown of lipids in this organ.

Menor eficacia de la lactancia materna como método contraceptivo en las poblaciones de altura: rol de la prolactina / Fewer eficacy of the breast-feeding as contraceptive method at high alttitude poblation

El presente estudio se ha llevado a cabo para determinar la tasa de embarazos durante la lactancia materna a nivel del mar y en la altura: así como los niveles de prolactina sérica durante la lactancia materna, y cuanto se modifican estos niveles de prolactina con la suplementación de la leche. La duración de la lactancia materna es similar en Lima, Cusco y Cerro de Pasco (11.4 meses), mientras que la paridad aumenta en función de la altitud en mujeres de la misma edad y que no usan métodos contraceptivos. El porcentaje de mujeres que se embarazan durante la lactancia se incrementa conforme se incrementa la altitud de residencia, siendo la probabilidad de embarazarse 6 veces más alta en Cerro de Pasco (4,340 m), que en Lima (150 m). La lactancia materna exclusiva es más frecuente en Cerro de Pasco (71 por ciento) que en Lima (55 por ciento), mientras que la frecuencia de lactadas diurnas y nocturnas es similar a nivel del mar y en la altura. El intervalo entre el primer y segundo nacimiento es menor a medida que se incrementa la altitud de residencia.

Modulation in activity of some epididymal glycosidases by prolactin.

Activity of glycosidases in the epididymis was influenced by several factors originating in the testis. Activities of all the three glycosidases studied viz., beta-glucosidase, beta-galactosidase and alpha-mannosidase were found to be significantly lower in the epididymis of orchidectomized animals than in sham operated rats. However, an enhanced activity of epididymal beta-galactosidase and alpha-mannosidase was noticed in prolactin treated orchidectomized rats compared to orchidectomized rats given vehicle alone. On the other hand, activity of these two enzymes in bromocriptine treated orchidectomized rats was even lower than that found in orchidectomized rats given vehicle. Neither prolactin nor bromocriptine treatment had any significant influence on the epididymal beta-glucosidase. The results suggest a selective but definite action of prolactin on epididymal glycosidases.

Effect of thyroxine, prolactin and adrenalin on the gravid ovaries of the frog Rana cyanophlyctis (Schn.).

The effects of thyroxine, prolactin and adrenalin on the gravid ovaries of R. cyanophlyctis were studied during late prebreeding period when vitellogenic growth of oocytes is complete or near completion. Specified doses of hormones were injected (ip) six days a week for one month. They were fed guppies ad libitum daily 6 days a week. Administration of 2 or 5 micrograms eltroxine (synthetic thyroxine) had no effect on ovaries or oviducts. Whereas, higher doses 8, 12 and 16 micrograms thyroxine caused significant (P < 0.05) decrease in the per cent weight of ovaries and oviducts. The mean diameter of the largest oocyte was also reduced significantly compared to the controls. There was an increase in the atresia of vitellogenic oocytes. Treatment with 100 micrograms prolactin also caused reduction in the per cent weight of ovaries and oviducts. However, mean diameter of the largest oocyte did not change significantly. Follicular atresia increased only moderately. Injection of 180 micrograms adrenalin bitartrate caused drastic decrease (P < 0.05) in the per cent weight of ovaries and oviducts and mean diameter of the largest oocyte. Many yolky follicles became atretic. The findings suggest that increase in the levels of thyroxine (beyond a certain limit), prolactin and adrenalin around the breeding period decrease the fecundity of the frog by inducing atresia of yolky oocytes. We conclude that the above hormones in excess quantities act as negative modifiers of ovarian activity in R. cyanophlyctis.

Effect of prolactin on accessory sex organs of prepuberal male albino rats: an in vitro study.

The in vitro studies on the effect of hypophysial gonadotropins (PRL, FSH, LH) on the maturation events of accessory sex organs in prepuberal male rats revealed that prolactin (PRL) alone has both direct as well as androgen mediated effect on the maturation activities. The effect of PRL is age dependent and it had higher activation on the gland system than the duct system.

Effects of prolactin and androgens on enzymes of carbohydrate metabolism in prostate of castrated bonnet monkeys Macaca radiata (Geoffroy).

Effects of prolactin (PRL), bromocriptine (Br), testosterone propionate (TP), dihydrotestosterone (DHT) and the combinations of these androgens with PRL/Br on the specific activities of caudal and cranial prostatic cellular enzymes involved in carbohydrate metabolism in castrated mature bonnet monkeys have been studied. Castration decreased all the enzymes studied such as hexokinase (HK), 6-phosphofructokinase (6-PFK), glyceraldehyde-3-phosphate dehydrogenase (G-3-PD), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) in the cranial and caudal prostates. PRL elevated the activities of all the enzymes above normal except G-3-PD of cranial lobe. In the caudal lobe, PRL brought back the activities of HK, PFK, PK, G-6-PD to normal and 6-PGD above normal except G-3-PD. TP/DHT treatment increased all the enzymes in both the lobes. PRL given along with TP/DHT further enhanced the androgen action with regard to HK, PK, G-6-PD and 6-PGD of cranial and PFK, G-3-PD, PK, G-6-PD and 6-PGD of caudal lobe. Br treatment did not produce any alteration of these enzymes in both the lobes. In the cranial lobe, during Br+TP/DHT treatment, the stimulating effects of androgen were unaffected on all the enzymes except PK. On the other hand in the caudal, the stimulatory effects of androgens were affected and the activities of HK, PFK, PK and 6-PGD were significantly decreased. The present results suggest that PRL has a direct as well as a synergistic action with androgens on enzymes of EMP and HMP shunt in the prostates of monkeys.

Influence of prolactin on pulmonary lipids in mature male bonnet monkey, Macaca radiata.

Administration of ovine prolactin (250 micrograms/kg body weight/day) for 30 days to mature male bonnet monkeys increased the pulmonary total lipids and total phospholipids. While total cholesterol did not show any appreciable change in PRL treated monkeys, the ratio of free: esterified cholesterols was altered with perceptible decrease in free cholesterol and increase in esterified cholesterol. Pulmonary tissue total glyceride glycerol concentration was diminished as a result of decreased monoacyl, diacyl and triacyl glycerols. Among phospholipid fractions, phosphatidylcholine and phosphatidylinositol were elevated while phosphatidic acid and cardiolipin were reduced. The data suggest that hyperprolactinemia interferes with pulmonary lipid metabolism in adult monkeys and modifies the ratio of cholesterol and phospholipid composition. Further, hyperprolactinemia appears to favour esterification of cholesterol with specific changes in phospholipid fractions. Thus, the present study demonstrates that hyperprolactinemia modifies the pattern of pulmonary neutral and phospholipid fractions and thereby is likely to affect the structure and function of mature primate lungs.

Studies on hormonal modulation of asparagine-linked glycoprotein biosynthesis in explant cultures of rat mammary gland.

Glucosidase I has been purified to homogeneity and polyclonal antibodies against the enzyme have been prepared. The anti-glucosidase I antibodies recognized a single band of 85 kDa on western blot at a dilution as high as 1:2000 and also inhibited the enzyme activity, suggesting the specificity of the antibodies. Con A-Sepharose binding experiment indicates that this enzyme itself is a high mannose type N-linked glycoprotein. The increase in the electrophoretic mobility of 85 kDa band following digestion with endoglycosidase H and F strengthened this observation. The presence of any O-linked sugar attached covalently to glucosidase I could not be detected by binding assays with O-linkage specific biotinylated lectins. The studies on developmental regulation suggest that the synthesis of glucosidase I is modulated with the ontogeny of the gland. Lactogenic hormones, viz. insulin, hydrocortisone and prolactin, appeared to regulate the synthesis of glucosidase I. The possible role of these hormones in the overall regulation of protein N-glycosylation has been discussed.

Improved prolactin crop-sac bioassay applying a morphometric approach

A pigeon crop-sac bioassay for prolactin was developed applying a morphometric approach for quantification of the epithelial cell proliferation induced by the hormone. A small male dove, indigenous from South America (Zenaida auriculata) previously castrated was used as experimental animal. The total dose of prolactin was divided into 4 daily injections administered systemically into the pectoral muscles. The number of the proliferated cell layers ysed as the bioassay end point was analyzedstatistically. This procedure yields a highly sensitive bioassay with an excellent precision index, particularly well suited for quantification of the different molecular forms of pituitary PRL

Prolactin effects on testicular interstitial cells in primary cultures

En el presente trabajo se determinaron los efectos de la prolactina (PRL) "in vitro" sobre la esteroidogénesis testicular, utilizando un cultivo primario de células intersticiales de rata inmadura. Las células fueron cultivadas en un medio químicamente definido, con el agregado de insulina y transferrina durante 2 días a 34§C. La producción de andrógenos durante el segundo día de cultivo resultó ser significativamente mayor que en el primer día (3 alfa-Androstandiol (3 alfa-Diol) 5.14 ñ 0.46 vs. 3.74 ñ 0.10 ng/microng ADN, p < 0.001); Testosterona (T) + Dihidrotestosterona (DHT) 0.40 ñ 0.04 vs. 0.34 ñ 0.03 ng/microng ADN, p < 0.001). La curva dosis respuesta para distintas dosis de hCG mostró un ED50= 2.5 mUI/ml. El efecto agudo de la PRL (10, 100 y 1000 ng/ml) sobre la producción basal de 3 alfa/Diol se evaluó luego de 45 h de cultivo. Sólo la dosis de PRL de 1000 ng/ml reveló diferencias significativas con respcto al control y dicho efecto pordría ser debido a su contaminación con LH. En otra serie de experimentos se evaluaron los efectos de menores dosis de la hormona a tiempos prolongados. La PRL (10 ng/ml), agregada durante todo el tiempo de cultivo, causó una inhibición significativa en la producción basal de 3 alfa-Diol, mientras que la respuesta a un estímulo máximo con hCG no varió. Cuando se determinó la producción de T+DHT se observó un cambio notorio en la relación T+DHT/Diol, tanto en condiciones basales (Control: 0.09 vs. PRL: 0.38) como ante el estímulo con hCG...

Effects of prolactin and androgens on seminal vesicular lipids of castrated mature bonnet monkeys Macaca radiata.

Effects of prolactin (PRL), bromocriptine (Br), testosterone propionate (TP), dihydrotestosterone (DHT) and the combination of these androgens with PRL/Br on the total lipid, total cholesterol, total glyceride glycerols, total phospholipid and their fractions in seminal vesicles of castrated mature monkeys were studied. Glyceride glycerols formed the major portion (50%) of total lipids in normal monkeys. Cholesterol and phospholipids were of equal share (25%). Esterified cholesterol formed major share (75%) of total cholesterol. Diacyl glycerol was the major (60%) glyceride glycerol and phosphatidyl choline and ethanolamine were the major phospholipid classes. Except triacyl glycerol castration markedly decreased all the lipid classes. PRL restored normal free and esterified cholesterol and phosphatidyl inositol but Br invariably decreased all the lipid classes. TP/DHT treatment stimulated the free and esterified cholesterol more than the control; it restored the normal glyceride glycerols. Phosphatidyl inositol, choline and ethanolamine were stimulated by androgens and other phospholipid classes were brought to normal. Addition of PRL + TP/DHT markedly increased esterified cholesterol, phosphatidyl inositol, choline, ethanolamine and phosphatidic acid. In all these aspects, Br counteracted the effects of androgens and PRL.