RESUMEN
In female rats, stimulation of the uterine cervix during mating induces two daily surges of prolactin. Inhibition of hypothalamic dopamine release and stimulation of oxytocin neurons in the paraventricular nucleus (PVN) are required for prolactin secretion. We aim to better understand how stimulation of the uterine cervix is translated into two daily prolactin surges. We hypothesize that noradrenergic neurons in the A1, A2, and locus coeruleus (LC) are responsible for conveying the peripheral stimulus to the PVN. In order to determine whether projections from these neurons to the PVN are activated by cervical stimulation (CS), we injected a retrograde tracer, Fluoro-Gold (FG), into the PVN of ovariectomized rats. Fourteen days after injection, animals were submitted to artificial CS or handling and perfused with a fixative solution. Brains were removed and sectioned from the A1, A2, and LC for c-Fos, tyrosine hydroxylase (TH), and FG triple-labeling using immunohistochemistry. CS increased the percentage of TH/FG+ double-labeled neurons expressing c-Fos in the A1 and LC. CS also increased the percentage of TH+ neurons expressing c-Fos within the A1 and A2, independent of their projections to the PVN. Our data reinforce the significant contributions of the A1 and A2 to carry sensory information during mating, and provide evidence of a functional pathway in which CS activates A1 and LC neurons projecting to the PVN, which is potentially involved in the translation of CS into two daily prolactin surges.
Asunto(s)
Cuello del Útero/inervación , Ritmo Circadiano/fisiología , Copulación/fisiología , Sistema Hipotálamo-Hipofisario/fisiología , Locus Coeruleus/fisiología , Plexo Lumbosacro/fisiología , Bulbo Raquídeo/fisiología , Vías Nerviosas/fisiología , Núcleo Hipotalámico Paraventricular/fisiología , Animales , Transporte Axonal , Femenino , Colorantes Fluorescentes , Lactotrofos/metabolismo , Locus Coeruleus/citología , Locus Coeruleus/metabolismo , Bulbo Raquídeo/citología , Bulbo Raquídeo/metabolismo , Proteínas del Tejido Nervioso/análisis , Vías Nerviosas/ultraestructura , Neuronas/química , Neuronas/metabolismo , Ovariectomía , Oxitocina/metabolismo , Prolactina/metabolismo , Proteínas Proto-Oncogénicas c-fos/análisis , Ratas , Ratas Sprague-Dawley , Estilbamidinas , Tirosina 3-Monooxigenasa/análisisRESUMEN
We have shown previously that an intravenous injection of oxytocin (OT) in ovariectomized (OVX) rats initiates a circadian rhythm of prolactin (PRL) secretion similar to that observed after cervical stimulation (CS). In this study, we investigated the pathway through which OT triggers the PRL rhythm. We first tested whether an intracerebroventricular injection of OT could trigger the PRL secretory rhythm. As it did not, we injected OT intravenously while an OT receptor antagonist was infused intravenously. This antagonist completely abolished the PRL surges, suggesting that a peripheral target of OT is necessary for triggering the PRL rhythm. We hypothesized that OT may induce PRL release, which would be transported into the brain and trigger the rhythm. In agreement with this, OT injection increased circulating PRL by 5 min. To test whether this acute increase in PRL release would induce the PRL rhythm, we compared the effect of intravenously administered thyrotropin-releasing hormone (TRH) and OT. Although TRH injection also increased PRL to a comparable level after 5 min, only OT-injected animals expressed the PRL secretory rhythm. Motivated by prior findings that bilateral resection of the pelvic nerve blocks CS-induced pseudopregnancy and OT-induced facilitation of lordosis, we then hypothesized that the OT signal may be transmitted through the pelvic nerve. In fact, OT injection failed to induce a PRL secretory rhythm in pelvic-neurectomized animals, suggesting that the integrity of the pelvic nerve is necessary for the systemic OT induction of the PRL secretory rhythm in OVX rats.
Asunto(s)
Ritmo Circadiano , Plexo Hipogástrico/efectos de los fármacos , Ovariectomía , Oxitocina/administración & dosificación , Adenohipófisis/efectos de los fármacos , Prolactina/metabolismo , Análisis de Varianza , Animales , Desnervación , Femenino , Plexo Hipogástrico/cirugía , Infusiones Intravenosas , Inyecciones Intraventriculares , Adenohipófisis/inervación , Adenohipófisis/metabolismo , Ratas , Ratas Sprague-Dawley , Hormona Liberadora de Tirotropina/administración & dosificación , Factores de TiempoRESUMEN
Although removal of dopamine inhibition is established as a major factor in prolactin (PRL) release, a large body of evidence suggests that hypothalamic oxytocin (OT) may serve as a PRL-releasing hormone in the rat. PRL release is modulated by estradiol (E2), which rises between diestrus and proestrus of the estrous cycle, causing a PRL surge in the afternoon of proestrus. Given that E2 strongly modulates OT actions in both central and peripheral tissues, OT action on lactotrophs might also be modulated by the stage of the estrous cycle. To test this hypothesis, we have monitored PRL release and intracellular calcium levels ([Ca(2+)](i)) induced by OT in pituitary lactotrophs obtained from female rats in either diestrus 1 or proestrus. We found that both secretory and [Ca(2+)](i) responses to OT are significantly increased in lactotrophs obtained on proestrus. Moreover, we show that these differences are due to an increase in both the number of OT-responding lactotrophs and the magnitude of their individual [Ca(2+)](i) responses. Both secretory and [Ca(2+)](i) responses were abolished by a specific OT antagonist. Finally, dose-dependent studies show that the increased PRL-releasing effect of OT on proestrus is significant over a wide range of concentrations, particularly those observed in hypophyseal portal plasma. These results suggest that the rising E2 titers that culminate on proestrus facilitate the stimulatory action of OT on lactotrophs and support the notion that OT is a PRL-releasing hormone with an important role in the production of the proestrous surge of PRL.
Asunto(s)
Ciclo Estral/metabolismo , Lactotrofos/metabolismo , Oxitocina/metabolismo , Animales , Área Bajo la Curva , Calcio/metabolismo , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Lactotrofos/efectos de los fármacos , Oxitocina/farmacología , Prolactina/metabolismo , Radioinmunoensayo , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND/AIMS: Cervical stimulation induces a circadian rhythm of prolactin secretion and antiphase dopamine release. The suprachiasmatic nucleus (SCN) controls this rhythm, and we propose that it does so through clock gene expression within the SCN. METHODS: To test this hypothesis, serial blood samples were taken from animals injected with an antisense deoxyoligonucleotide cocktail for clock genes (generated against the 5' transcription start site and 3' cap site of per1, per2, and clock mRNA) or with a random-sequence deoxyoligonucleotide in the SCN. To determine whether disruption of clock genes in the SCN compromises the neural mechanism controlling prolactin secretion, we sacrificed another group of rats (under the same treatments) at 12.00 or 17.00 h. Dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) were measured using HPLC/electrochemical detection in the median eminence as well as the intermediate and the neural lobe of the pituitary gland, and the DOPAC:dopamine ratio was used as an index of dopamine activity. Vasoactive intestinal polypeptide (VIP) content was determined in tissue punches of the SCN and paraventricular nucleus (PVN), an SCN efferent. RESULTS: Treatment with clock gene antisense deoxyoligonucleotide cocktail abolished both the diurnal and nocturnal prolactin surges induced by cervical stimulation. This treatment abolished the antiphase relationship established by cervical stimulation between dopamine neuronal activity and prolactin secretion. Also, VIP content increased in the SCN and decreased in the PVN. CONCLUSION: These results suggest that the SCN clock determines the circadian rhythm of prolactin secretion in cervically stimulated rats by regulating dopamine neuronal activity and VIP inputs to the PVN.
Asunto(s)
Relojes Biológicos/fisiología , Ritmo Circadiano/fisiología , Dopamina/metabolismo , Prolactina/sangre , Péptido Intestinal Vasoactivo/metabolismo , Animales , Proteínas CLOCK/genética , Proteínas CLOCK/metabolismo , Femenino , Técnicas de Silenciamiento del Gen , Ovariectomía , Núcleo Hipotalámico Paraventricular/fisiología , Proteínas Circadianas Period/genética , Proteínas Circadianas Period/metabolismo , Ratas , Ratas Sprague-Dawley , Núcleo Supraquiasmático/fisiologíaRESUMEN
Pro-opiomelanocortin (POMC) is processed to adrenocorticotropic hormone (ACTH) and beta-lipotropin in corticotropes of the anterior lobe, and to alpha-melanocyte-stimulating hormone (alpha-MSH) and beta-endorphin in melanotropes of the intermediate lobe (IL) of the pituitary gland. While ACTH secretion is predominantly under the stimulatory influence of the hypothalamic factors, hormone secretion of the IL is tonically inhibited by neuroendocrine dopamine (NEDA) neurons. Lobe-specific POMC processing is not absolute. For example, D(2) type DA receptor (D2R)-deficient mice have elevated plasma ACTH levels, although it is known that corticotropes do not express D2R(s). Moreover, observations that suckling does not influence alpha-MSH release, while it induces an increase in plasma ACTH is unexplained. The aim of the present study was to investigate the involvement of the NEDA system in the regulation of ACTH secretion and the participation of the IL in ACTH production in lactating rats. Untreated and estradiol (E(2))-substituted ovariectomized (OVX) females were also studied. The concentration of ACTH in the IL was higher in lactating rats than in OVX rats, while the opposite change in alpha-MSH level of the IL was observed. DA levels in the IL and the neural lobe were lower in lactating rats than in OVX rats. Suckling-induced ACTH response was eliminated by pretreatment with the DA receptor agonist, bromocriptine (BRC). Inhibition of DA biosynthesis by alpha-methyl-p-tyrosine (alphaMpT) and blockade of D2R by domperidone (DOM) elevated plasma ACTH levels, but did not influence plasma alpha-MSH levels in lactating rats. The same drugs had opposite effects in OVX and OVX + E(2) animals. In lactating mothers, BRC was able to block ACTH responses induced by both alphaMpT and DOM. Surgical denervation of the IL elevated basal plasma levels of ACTH. Taken together, these data indicate that melanotropes synthesize ACTH during lactation and its release from these cells is regulated by NEDA neurons.
Asunto(s)
Hormona Adrenocorticotrópica/metabolismo , Dopamina/metabolismo , Lactancia/fisiología , Hipófisis/fisiología , Ácido 3,4-Dihidroxifenilacético/metabolismo , Animales , Bromocriptina/farmacología , Domperidona/farmacología , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Antagonistas de los Receptores de Dopamina D2 , Inhibidores Enzimáticos/farmacología , Estradiol/metabolismo , Femenino , Lactancia/sangre , Lactancia/efectos de los fármacos , Ovariectomía , Hipófisis/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Receptores de Dopamina D2/agonistas , alfa-MSH/sangre , alfa-MSH/metabolismo , alfa-Metiltirosina/farmacologíaRESUMEN
Artificial copulomimetic cervical stimulation (CS) induces an immediate release of oxytocin (OT) and prolactin (PRL) followed by a daily PRL rhythm characterized by nocturnal and diurnal surges. Although we have shown that the initial release of PRL is induced by the immediate release of OT, we tested whether the PRL that is released in response to CS is responsible for the initiation and maintenance of the subsequent PRL surges. Thus, we injected OVX rats centrally or peripherally with ovine PRL (oPRL) at 2200 h. Central oPRL induced PRL surges in OVX rats that were similar in size and timing to those of CS rats, whereas peripheral oPRL induced surges that were of smaller amplitude and delayed. We then infused a PRL antagonist (S179D, 0.1 ng/h) centrally into OVX and OVX-CS rats and measured the release of endogenous PRL and the activity of neuroendocrine dopaminergic neurons. Central infusion of S179D did not influence basal PRL secretion in OVX rats but prevented the expression of the CS-induced PRL surges and the accompanying noontime increase of CS-induced dopaminergic activity when continued for 3 d. However, central infusion of S179D only on the day of CS did not prevent the daily rhythm of PRL surges. These results demonstrate that PRL acts centrally to induce the PRL rhythm and that PRL in the brain is essential for the maintenance but not for the initiation of the CS-induced rhythmic PRL surges.
Asunto(s)
Cuello del Útero/fisiología , Prolactina/metabolismo , Prolactina/fisiología , Animales , Ritmo Circadiano/fisiología , Estimulación Eléctrica , Femenino , Ovariectomía , Oxitocina/metabolismo , Prolactina/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Prolactina/antagonistas & inhibidoresRESUMEN
In female rats, estradiol (E(2)) and suckling induce prolactin (PRL) secretion. This involves inhibition of hypothalamic dopaminergic tone and stimulation by a PRL-releasing hormone, possibly oxytocin (OT). Infusing an OT antagonist (OTA) i.v., we evaluated the role of OT on suckling- and E(2)-induced PRL secretion. Three days after parturition at 0900 h, lactating dams were fitted with 24-h osmotic minipumps filled with saline or OTA. On d 5 of lactation, pups were separated from their dams for 6 h. Immediately or 20 min after the resumption of suckling, dam trunk blood was collected. Also, ovariectomized (OVX) rats were treated with E(2) (OVE) and OTA at 1000 h on d 1. Blood samples were obtained from 1300 to 2100 h on d 2 for PRL measurements. Additionally, OVX rats were evaluated on d 2 after receiving progesterone (P(4)). OTA blocked suckling and E(2)-induced release of PRL but not that induced by E(2)+P(4). Pups from treated dams failed to gain weight when allowed to nurse for 20 min on d 5 but gained more than 7 g when nursed on d 7 of lactation, indicating that the OTA was active 48 h later. Western blot analysis showed that E(2) treatment increased OT receptors in the anterior pituitary when compared with OVX animals. No further increase was observed in response to the P(4), suggesting that the enhancing effect of P(4) on E(2)-induced PRL release may act through mechanisms independent of OT. These data demonstrate the role of OT in the control of suckling and steroid-induced PRL secretion.
Asunto(s)
Estradiol/farmacología , Ornipresina/análogos & derivados , Oxitocina/antagonistas & inhibidores , Progesterona/farmacología , Prolactina/metabolismo , Conducta en la Lactancia/efectos de los fármacos , Animales , Animales Recién Nacidos , Animales Lactantes , Femenino , Bombas de Infusión , Lactancia/efectos de los fármacos , Ornipresina/administración & dosificación , Ornipresina/farmacología , Ovariectomía/veterinaria , Oxitocina/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
In female rats, estradiol is responsible for a circadian secretory prolactin (PRL) pattern which requires an intact suprachiasmatic nucleus (SCN). SCN outputs involved in this secretory profile remain elusive. Because oxytocin has been proposed to stimulate PRL secretion, we investigated whether the projections of vasoactive intestinal polypeptide (VIP) from the SCN to neurons producing oxytocin in the paraventricular and periventricular nuclei (PVN and PeVN, respectively) are responsible for timing PRL surges induced by estradiol (E(2)). E(2)-treated ovariectomized rats received an injection of antisense or random-sequence oligodeoxynucleotide for VIP in the SCN and blood samples were taken for PRL measurements by radioimmunoassay. Additionally, the percentage of oxytocin-positive neurons immunoreactive to FOS-related antigens was determined in the PVN and PeVN, as an index of neuronal activity. In the PVN, oxytocinergic neuronal activity increased in the early evening regardless of E(2) treatment, whereas E(2) induced an increase of activity in the PeVN. VIP antisense attenuated this increase observed in both neuronal populations. Additionally, in the PeVN, VIP antisense advanced this increase by 2 h (from 19:00 h to 17:00 h). This same effect was observed in the PRL surge that occurred at 17:00 h in the VIP antisense injected animals. Thus, the SCN influences the precise timing of the E(2)-induced PRL surge via VIP projections to oxytocinergic neurons of the PVN and PeVN.
Asunto(s)
Estradiol/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Oxitocina/metabolismo , Prolactina/metabolismo , Péptido Intestinal Vasoactivo/farmacología , Análisis de Varianza , Animales , Ritmo Circadiano/efectos de los fármacos , Interacciones Farmacológicas , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Oligodesoxirribonucleótidos Antisentido/farmacología , Ovariectomía , Núcleo Hipotalámico Paraventricular/fisiología , Radioinmunoensayo/métodos , Ratas , Ratas Sprague-Dawley , Núcleo Supraquiasmático/citología , Factores de TiempoRESUMEN
Models of bursting in single cells typically include two subsystems with different timescales. Variations in one or more slow variables switch the system between a silent and a spiking state. We have developed a model for bursting in the pituitary lactotroph that does not include any slow variable. The model incorporates fast, noninactivating calcium and potassium currents (the spike-generating mechanism), as well as the fast, inactivating A-type potassium current (I(A)). I(A) is active only briefly at the beginning of a burst, but this brief impulse of I(A) acts as a burst trigger, injecting the spike trajectory close to an unstable steady state. The spiraling of the trajectory away from the steady state produces a period of low-amplitude spiking typical of lactotrophs. Increasing the conductance of A-type potassium current brings the trajectory closer to the unstable steady state, increasing burst duration. However, this also increases interburst interval, and for larger conductance values, all activity stops. To our knowledge, this is the first example of a physiologically based, single-compartmental model of bursting with no slow subsystem.
Asunto(s)
Potenciales de Acción/fisiología , Lactotrofos/fisiología , Modelos Neurológicos , Canales de Potasio/fisiología , Animales , Calcio/metabolismo , Potasio/metabolismo , Tiempo de Reacción/fisiologíaRESUMEN
The nature of the circadian signal from the suprachiasmatic nucleus (SCN) required for prolactin (PRL) surges is unknown. Because the SCN neuronal circadian rhythm is determined by a feedback loop of Period (Per) 1, Per2, and circadian locomotor output cycles kaput (Clock) gene expressions, we investigated the effect of SCN rhythmicity on PRL surges by disrupting this loop. Because lesion of the locus coeruleus (LC) abolishes PRL surges and these neurons receive SCN projections, we investigated the role of SCN rhythmicity in the LC neuronal circadian rhythm as a possible component of the circadian mechanism regulating PRL surges. Cycling rats on proestrous day and estradiol-treated ovariectomized rats received injections of antisense or random-sequence deoxyoligonucleotide cocktails for clock genes (Per1, Per2, and Clock) in the SCN, and blood samples were taken for PRL measurements. The percentage of tyrosine hydroxylase-positive neurons immunoreactive to Fos-related antigen (FRA) was determined in ovariectomized rats submitted to the cocktail injections and in a 12:12-h light:dark (LD) or constant dark (DD) environment. The antisense cocktail abolished both the proestrous and the estradiol-induced PRL surges observed in the afternoon and the increase of FRA expression in the LC neurons at Zeitgeber time 14 in LD and at circadian time 14 in DD. Because SCN afferents and efferents were probably preserved, the SCN rhythmicity is essential for the magnitude of daily PRL surges in female rats as well as for LC neuronal circadian rhythm. SCN neurons therefore determine PRL secretory surges, possibly by modulating LC circadian neuronal activity.
Asunto(s)
Ritmo Circadiano/fisiología , Ciclo Estral/fisiología , Locus Coeruleus/fisiología , Prolactina/metabolismo , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Núcleo Supraquiasmático/fisiología , Transactivadores/fisiología , Animales , Proteínas CLOCK , Ritmo Circadiano/genética , Ingestión de Líquidos/fisiología , Estradiol/farmacología , Proteínas del Ojo/fisiología , Femenino , Inmunohistoquímica , Locus Coeruleus/metabolismo , Neuronas/metabolismo , Neuronas/fisiología , Oligodesoxirribonucleótidos Antisentido/farmacología , Ovariectomía , Proteínas Circadianas Period , Proteínas Proto-Oncogénicas c-fos/fisiología , Ratas , Ratas Sprague-Dawley , Transactivadores/genéticaRESUMEN
Cervical stimulation induces two daily rhythmic prolactin surges, nocturnal and diurnal, which persist for several days. We have shown that a bolus injection of oxytocin initiates a similar prolactin rhythm, which persists despite low levels of oxytocin after injection. This suggests that oxytocin may trigger the cervical stimulation-induced rhythmic prolactin surges. To investigate this hypothesis, we infused an oxytocin antagonist that does not cross the blood-brain barrier for 24 h before and after cervical stimulation and measured serum prolactin. We also measured dopaminergic neuronal activity because mathematical modeling predicted that this activity would be low in the presence of the oxytocin antagonist. We thus tested this hypothesis by measuring dopaminergic neuronal activity in the tuberoinfundibular, periventricular hypophyseal, and tuberohypophyseal dopaminergic neurons. Infusion of oxytocin antagonist before cervical stimulation abolished prolactin surges, and infusion of oxytocin antagonist after cervical stimulation abolished the diurnal and significantly decreased the nocturnal surges of prolactin. The rhythmic prolactin surges returned after the clearance of the oxytocin antagonist. Hypothalamic dopaminergic activity was elevated in antiphase with prolactin surges, and the antiphase elevation was abolished by the oxytocin antagonist in the tuberoinfundibular and tuberohypophyseal dopaminergic neurons, consistent with the mathematical model. These findings suggest that oxytocin is a physiologically relevant prolactin-releasing factor. However, the cervical stimulation-induced prolactin surges are maintained even in the absence of oxytocin actions at the lactotroph, which strongly suggests the maintenance of prolactin surges are not dependent upon oxytocin actions at the pituitary gland.
Asunto(s)
Cuello del Útero/fisiología , Lactotrofos/metabolismo , Ovariectomía , Oxitocina/fisiología , Prolactina/metabolismo , Animales , Ritmo Circadiano , Dopamina/metabolismo , Estimulación Eléctrica , Femenino , Eminencia Media/citología , Eminencia Media/metabolismo , Modelos Biológicos , Neuronas/fisiología , Ornipresina/análogos & derivados , Ornipresina/farmacología , Oxitocina/antagonistas & inhibidores , Adenohipófisis Porción Intermedia/citología , Adenohipófisis Porción Intermedia/metabolismo , Neurohipófisis/citología , Neurohipófisis/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Dopamine (DA) released from the hypothalamus tonically inhibits pituitary lactotrophs. DA (at micromolar concentration) opens potassium channels, hyperpolarizing the lactotrophs and thus preventing the calcium influx that triggers prolactin hormone release. Surprisingly, at concentrations approximately 1000 lower, DA can stimulate prolactin secretion. Here, we investigated whether an increase in a K+ current could mediate this stimulatory effect. We considered the fast K+ currents flowing through large-conductance BK channels and through A-type channels. We developed a minimal lactotroph model to investigate the effects of these two currents. Both IBK and IA could transform the electrical pattern of activity from spiking to bursting, but through distinct mechanisms. IBK always increased the intracellular Ca2+ concentration, while IA could either increase or decrease it. Thus, the stimulatory effects of DA could be mediated by a fast K+ conductance which converts tonically spiking cells to bursters. In addition, the study illustrates that
Asunto(s)
Dopamina/farmacología , Lactotrofos/efectos de los fármacos , Hipófisis/citología , Canales de Potasio Calcio-Activados/fisiología , Prolactina/metabolismo , Animales , Calcio/metabolismo , Activación del Canal Iónico/efectos de los fármacos , Activación del Canal Iónico/fisiología , Modelos Biológicos , Potasio/farmacología , Canales de Potasio Calcio-Activados/efectos de los fármacosRESUMEN
The endothelins are a family of hormones that have a biphasic action on pituitary lactotrophs. The initial effect is stimulatory, followed later by inhibition that persists long after the agonist has been removed. Recent research has uncovered several G protein pathways that mediate these effects.
Asunto(s)
Endotelinas/fisiología , Hipófisis/metabolismo , Prolactina/metabolismo , Animales , Calcio/metabolismo , Endotelina-1/fisiología , Exocitosis , Proteínas de Unión al GTP/fisiología , Humanos , Modelos Biológicos , Ratas , Transducción de SeñalRESUMEN
Mating or vaginocervical stimulation [copulatory stimulus (CS)] induces two daily surges of the hormone prolactin (PRL) in rats. This unique secretory pattern of PRL surges is characteristic for the first half of pregnancy and is also present in ovariectomized (OVX) rats. Studies have shown that CS additionally provokes an acute release of the hormone oxytocin (OT). In this study, we tested whether a single injection of OT (iv) is sufficient to initiate the PRL secretion pattern of OVX/CS rats. Furthermore, we measured the 24-h profile of dopamine (DA) content in the anterior lobe of the pituitary gland, because DA is the major inhibitory factor of PRL secretion. The results indicated that a single injection of OT induces a PRL secretory rhythm and a DA release pattern similar to that initiated by CS. Immunocytochemical investigation showed that particular OTergic neurons in the hypothalamus express receptors for PRL, as well as for vasoactive intestinal polypeptide, which indicates an involvement in generating the PRL rhythm and entraining it to the ambient photoperiod. On the basis of this study, we suggest that the PRL-DA inhibitory feedback loop between lactotrophs and DAergic neurons plays a crucial role in generating the oscillatory PRL secretion pattern in CS rats. A timing signal, likely provided by the hypothalamic suprachiasmatic nucleus, entrains the autonomous PRL oscillation to a particular time of day. Mathematical modeling was used to illustrate the proposed network function. The experimental results further suggest an additional feedback mechanism in which certain hypothalamic OTergic neurons are influenced by PRL.
Asunto(s)
Copulación/fisiología , Oxitocina/farmacología , Adenohipófisis/efectos de los fármacos , Prolactina/metabolismo , Animales , Ritmo Circadiano/efectos de los fármacos , Ritmo Circadiano/fisiología , Dopamina/sangre , Dopamina/fisiología , Retroalimentación , Femenino , Inmunohistoquímica , Modelos Biológicos , Ovariectomía , Adenohipófisis/metabolismo , Prolactina/sangre , Prolactina/fisiología , Ratas , Ratas Sprague-Dawley , Receptores de Prolactina/metabolismo , Receptores de Tipo II del Péptido Intestinal Vasoactivo/metabolismoRESUMEN
For the first 10 days of pregnancy and the first 12 days of pseudopregnancy, the secretion of prolactin (PRL) from pituitary lactotrophs is rhythmic, with two surges/day. This rhythm can also be triggered by bolus injection of oxytocin (OT). We describe a mathematical model for the initiation, maintenance, and termination of the OT-induced PRL rhythm. In our model, the mechanism for this circadian rhythm is mutual interaction between lactotrophs and neuroendocrine dopamine (DA) neurons. This rhythm is, under normal lighting conditions, entrained by the suprachiasmatic nucleus (SCN) but persists in the absence of input from the SCN. We postulate that OT injection triggers the rhythm by activating a population of bistable hypothalamic neurons that innervate and inhibit DA neurons. The bistable nature of these neurons allows them to act as a memory device, maintaining the rhythm long after OT has been cleared from the blood. The mechanism for this memory device and the arguments supporting it are detailed with computer simulations. Finally, we consider potential targets for a rhythm-terminating factor and make predictions that may be used to determine which mechanism is operational in terminating the OT- or mating-induced PRL rhythm.
Asunto(s)
Ritmo Circadiano/fisiología , Copulación/fisiología , Modelos Biológicos , Prolactina/metabolismo , Núcleo Supraquiasmático/fisiología , Animales , Simulación por Computador , Dopamina/fisiología , Retroalimentación/fisiología , Femenino , Neuronas/fisiología , RatasRESUMEN
The endothelins are a family of hormones that have a biphasic action on pituitary lactotrophs. The initial effect is stimulatory, followed later by inhibition that persists long after the agonist has been removed. Recent research has uncovered several G protein pathways that mediate these effects.
Asunto(s)
Endotelinas/fisiología , Proteínas de Unión al GTP/fisiología , Adenohipófisis/efectos de los fármacos , Prolactina/metabolismo , Receptores de Endotelina/fisiología , Transducción de Señal/fisiología , Adenilil Ciclasas/fisiología , Animales , Señalización del Calcio/fisiología , AMP Cíclico/fisiología , Dopamina/fisiología , Endotelina-1/farmacología , Endotelina-1/fisiología , Endotelinas/genética , Exocitosis/fisiología , Subunidades alfa de la Proteína de Unión al GTP/fisiología , Proteínas de Unión al GTP/clasificación , Humanos , Fosfatos de Inositol/fisiología , Modelos Biológicos , Adenohipófisis/citología , Adenohipófisis/metabolismo , Canales de Potasio Calcio-Activados/fisiología , Ratas , Receptor de Endotelina A/efectos de los fármacos , Receptor de Endotelina A/fisiología , Receptores de Endotelina/efectos de los fármacos , Proteínas SNARE/fisiología , Fosfolipasas de Tipo C/fisiologíaRESUMEN
Oscillations of gene expression and physiological activity in suprachiasmatic nucleus (SCN) neurons result from autoregulatory feedback loops of circadian clock gene transcription factors. In the present experiment, we have determined the pattern of PERIOD1 (PER1), PERIOD2 (PER2), and CLOCK expression within neuroendocrine dopaminergic (DAergic) neurons (NDNs) of ovariectomized (OVX) rats. We have also determined the effects of per1, per2, and clock mRNA knockdown in the SCN with antisense deoxyoligonucleotides (AS-ODN) on DA release from NDNs. Diurnal rhythms of PER1 and PER2 expression in tuberoinfundibular DAergic (TIDA) and periventricular hypophyseal DAergic (PHDA) neurons, peaked at circadian time (CT)18 and CT12, respectively. Rhythms of PER1 expression in tuberhypophyseal neuroendocrine DAergic (THDA) neurons were undetectable. Rhythms of PER2 expression were found in all three populations of NDNs, with greater levels of PER2 expression between CT6 and CT12. AS-ODN injections differentially affected DA turnover in the axon terminals of the median eminence (ME), neural lobe (NL) and intermediate lobe (IL) of the pituitary gland, resulting in a significant decrease in DA release in the early subjective night in the ME (TIDA), a significant increase in DA release at the beginning of the day in the IL (PHDA), and no effect in the NL (THDA). AS-ODN-treatment induced a rhythm of DA concentration in the anterior lobe, with greater DA levels in the middle of the day. These data suggest that clock gene expression, particularly PER1 and PER2, within NDNs may act to modulate diurnal rhythms of DA release from NDNs in the OVX rat.
Asunto(s)
Neuronas/fisiología , Sistemas Neurosecretores/anatomía & histología , Sistemas Neurosecretores/fisiología , Transactivadores/genética , Factores de Transcripción ARNTL , Animales , Animales Modificados Genéticamente , Elementos sin Sentido (Genética) , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Proteínas CLOCK , Proteínas de Ciclo Celular , Corticosterona/metabolismo , Señales (Psicología) , Conducta de Ingestión de Líquido/fisiología , Electroforesis en Gel de Poliacrilamida , Femenino , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Núcleos Talámicos de la Línea Media/metabolismo , Sistemas Neurosecretores/citología , Proteínas Nucleares/genética , Ovariectomía , Proteínas Circadianas Period , Prolactina/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Núcleo Supraquiasmático/fisiología , Factores de Transcripción/genéticaRESUMEN
Prairie (Microtus ochrogaster) and meadow voles (M. pennsylvanicus) are closely related species that differ in life strategy and social behaviors, and thus provide an excellent comparative model for the study of neuronal and hormonal mechanisms underlying behavior. In the present study using the elevated plus maze (EPM) test, we found that male prairie voles entered the open arms of the EPM more and remained there longer, and showed a higher level of overall locomotor activity than did male meadow voles. In addition, two weeks of social isolation induced an increase in open arm entries in prairie, but not meadow, voles. Prairie voles also had a higher level of circulating corticosterone compared to meadow voles, and the EPM test increased circulating corticosterone in prairie voles. Finally, social isolation coupled with the EPM test influenced Fos-immunoreactive expression in several brain areas, including the medial preoptic area, ventromedial hypothalamus, amygdala, and prefrontal cortex differently between the two species. Together, these data indicate a neural circuit involved in mediating anxiety-associated behavior in voles, and that the functioning of this circuit is influenced by social environment differently between social and non-social species.
Asunto(s)
Ansiedad/fisiopatología , Arvicolinae/clasificación , Arvicolinae/fisiología , Aislamiento Social/psicología , Animales , Ansiedad/metabolismo , Recuento de Células/métodos , Corticosterona/sangre , Inmunohistoquímica/métodos , Masculino , Aprendizaje por Laberinto/fisiología , Actividad Motora/fisiología , Proteínas Proto-Oncogénicas c-fos/metabolismo , Radioinmunoensayo/métodos , Especificidad de la Especie , Factores de TiempoRESUMEN
Dopamine (DA) is the primary inhibitor of prolactin (PRL) secretion. Three populations of neuroendocrine dopaminergic neurons (NDNs) designated tuberoinfundibular (TIDA), tuberohypophyseal (THDA) and periventricular hypophyseal DAergic (PHDA) neurons regulate PRL secretion. Given that ovarian steroids modulate both DA release and PRL secretion independently, we characterized the role of steroid hormones in coupling rhythmic NDN activity and PRL secretion. OVX rats under a standard 12:12 L:D cycle (L:D), constant dark (DD), or a 6-h phase-delayed L:D cycle (pdL:D) were treated with Estradiol-17beta (E) or E and Progesterone (E+P). NDN activity, defined by DA:DOPAC ratio in nerve terminals, was determined by HPLC-EC. E or E+P stimulated PRL surges in L:D that persisted under DD. In TIDA neurons, E or E+P treatment reduced the amount of DA released under L:D and DD and advanced the rhythm of DA turnover. E and E+P treatment reduced THDA and PHDA neuron activity under L:D, but did not affect these rhythms under DD. Circadian rhythms of PRL, corticosterone and DA turnover in NDN terminals from steroid treated rats entrained to a pdL:D cycle within 7 days. Therefore, ovarian steroids differentially adjust the timing and magnitude of NDN activity to facilitate coupling of DA release and PRL secretion.
Asunto(s)
Ritmo Circadiano/fisiología , Dopamina/metabolismo , Hormonas Esteroides Gonadales/metabolismo , Sistemas Neurosecretores/metabolismo , Ovario/metabolismo , Animales , Ritmo Circadiano/efectos de los fármacos , Corticosterona/sangre , Estradiol/metabolismo , Estradiol/farmacología , Femenino , Hormonas Esteroides Gonadales/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Sistemas Neurosecretores/efectos de los fármacos , Ovariectomía , Ovario/efectos de los fármacos , Progesterona/metabolismo , Progesterona/farmacología , Prolactina/sangre , Ratas , Ratas Sprague-DawleyRESUMEN
Prolactin (PRL) is secreted from lactotrophs of the anterior pituitary gland of rats in a unique pattern in response to uterine cervical stimulation (CS) during mating. Surges of PRL secretion occur in response to relief from hypothalamic dopaminergic inhibition and stimulation by hypothalamic releasing neurohormones. In this study, we characterized the role of oxytocin (OT) in this system and the involvement of vasoactive intestinal polypeptide (VIP) from the suprachiasmatic nucleus (SCN) in controlling OT and PRL secretion of CS rats. The effect of OT on PRL secretion was demonstrated in cultured lactotrophs showing simultaneous enhanced secretion rate and increased intracellular Ca(2+). Neurosecretory OT cells of the hypothalamic paraventricular nucleus that express VIP receptors were identified by using immunocytochemical techniques in combination with the retrogradely transported neuronal tracer Fluoro-Gold (iv injected). OT measurements of serial blood samples obtained from ovariectomized (OVX) CS rats displayed a prominent increase at the time of the afternoon PRL peak. The injection of VIP antisense oligonucleotides into the SCN abolished the afternoon increase of OT and PRL in CS-OVX animals. These findings suggest that VIP from the SCN contributes to the regulation of OT and PRL secretion in CS rats. We propose that in CS rats the regulatory mechanism(s) for PRL secretion comprise coordinated action of neuroendocrine dopaminergic and OT cells, both governed by the daily rhythm of VIP-ergic output from the SCN. This hypothesis is illustrated with a mathematical model.