RESUMO
Two experiments evaluated prebiotics added to feed on the recovery of Salmonella in broilers during grow-out and processing. In Experiment 1, "seeder" chicks were inoculated with Salmonella Typhimurium and placed with penmates. Treatments were: basal control diet, added 0.3% bamboo charcoal, 0.6% bamboo charcoal, or 0.12% Aromabiotic (medium chain fatty acids). The ceca from seeders and penmates were sampled to confirm Salmonella colonization at 3, 4, and 6 wk, and pen litter was sampled weekly. At 3 wk, charcoal fed chicks had significantly lower cecal recovery (37% lower) of Salmonella via direct plating but no differences at wk 4 or 6. At 6 wk, broilers fed Aromabiotic had no recovery of Salmonella from ceca with direct plating and significantly, 18%, lower recovery with enrichment. In Experiment 2, the treatments were: basal control diet, added 0.3% bamboo charcoal, 0.3% activated bamboo charcoal, or 0.3% pine charcoal. At placement, 2 seeders were challenged with Salmonella and commingled with penmates and ceca sampled at 1 and 2 wk, and ceca from 5 penmates/pen at 3 to 6 wk. Weekly, the pH of the crop and duodenum was measured from 1 penmate/pen and the litter surface sampled. At the end of grow-out broilers were processed. Results showed that penmates had colonized at 1 and 2 wk. Cecal Salmonella showed no differences except at 4 wk, when activated bamboo charcoal had a 18% lower recovery of Salmonella (enrichment) compared to the control (88%). Similar to Experiment 1, the recovery of Salmonella from the litter was not significantly different among treatments, however an overall decrease in recovery by 4 wk with direct plating reoccurred. The pH of the duodenum and the crop were not different among treatments. Crop pH (6.0) for all treatments were significantly higher at wk 1 compared to wk 2 to 6. Charcoals had minimal effect on Salmonella recovery in the ceca, but following defeathering, broilers fed charcoals had significantly lower Salmonella recovery from breast skin (charcoals 5+/60 compared to control 8+/20). While the addition of charcoals to broilers feed did not significantly affect Salmonella recovery during production (from litter or ceca samples) there was a lower Salmonella recovery from breast skin following scalding and defeathering.
Assuntos
Carvão Vegetal/química , Dieta/veterinária , Ácidos Graxos/metabolismo , Doenças das Aves Domésticas/prevenção & controle , Prebióticos , Salmonelose Animal/prevenção & controle , Salmonella typhimurium/efeitos dos fármacos , Ração Animal/análise , Animais , Papo das Aves/química , Duodeno/química , Concentração de Íons de Hidrogênio , Masculino , Pinus/química , Poaceae/química , Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Pele/microbiologiaRESUMO
Prolactin and oxytocin are important reproductive hormones implicated in several common adaptive functions during pregnancy, pseudopregnancy and lactation. Recently, extracellular recordings of supraoptic neurones have shown that prolactin may modulate the electrical activity of oxytocinergic neurones. However, no study has been conducted aiming to establish whether prolactin directly influences this activity in oxytocinergic paraventricular neurones. In the present study, we addressed this question by studying the effects of prolactin on the electrical activity and voltage-current relationship of identified paraventricular neurones in rat brain slices. Whole-cell recordings were obtained and neurones were classified on the basis of their morphological and electrophysiological fingerprint (i.e. magnocellular or parvicellular) and neuropeptide phenotype (i.e. oxytocinergic or non-oxytocinergic). We report that prolactin elicited a hyperpolarising current in 37% of the neurones in this nucleus, of which the majority (67%) were identified as putative magnocellular oxytocin neurones and the reminder (33%) were regarded as oxytocin-negative, parvicellular neuroendocrine neurones. Our results suggest that, in addition to the well-established negative feedback loop between prolactin-secreting lactotrophs and dopaminergic neurones in the arcuate nucleus, an inhibitory feedback loop also exists between lactotrophs and oxytocinergic paraventricular neurones.
Assuntos
Potenciais de Ação/fisiologia , Neurônios/fisiologia , Ocitocina/metabolismo , Núcleo Hipotalâmico Paraventricular/fisiologia , Prolactina/fisiologia , Animais , Imuno-Histoquímica , Neurônios/metabolismo , Núcleo Hipotalâmico Paraventricular/citologia , Técnicas de Patch-Clamp , RatosRESUMO
Hormone secretion often occurs in a pulsatile manner. In this review, we discuss two rhythms of in vivo prolactin release in female rats and the ongoing research that we and others have performed aiming to understand the mechanisms underlying them. The peptide hormone oxytocin appears to play an important role in both rhythms. One rhythm occurs during the first half of pregnancy, but can also be induced in ovariectomised rats. This is characterised by a circadian pattern with two prolactin surges per day. Two methods for triggering this rhythm are discussed, each utilising a unique physiological pathway that includes oxytocin action, presumably on pituitary lactotrophs. The second rhythm occurs during the oestrous cycle and is characterised by a surge of prolactin on the afternoon of pro-oestrus. We discuss recent findings that oxytocin is more effective at stimulating prolactin release from lactotrophs taken from animals on the afternoon of pro-oestrus than from those of animals on the morning of dioestrus 1, raising the possibility that this hormone plays a physiological role in the regulation of prolactin secretion during the oestrous cycle.
Assuntos
Ritmo Circadiano/fisiologia , Ocitocina/metabolismo , Prolactina/metabolismo , Animais , Colo do Útero/metabolismo , Ritmo Circadiano/efeitos dos fármacos , Ciclo Estral/fisiologia , Feminino , Modelos Teóricos , Ocitocina/farmacologia , Gravidez , Ratos , Receptores de Ocitocina/antagonistas & inibidores , Receptores de Ocitocina/metabolismoRESUMO
Broiler breeder hens are typically provided a restricted amount of feed once a day. This feed is rapidly consumed; therefore, the hens fast for an extended period of time before their next feeding. In the current research, the effects on reproductive performance of implementing a twice-a-day vs. a once-a-day feeding program after photostimulation were investigated. Pullets and cockerels were reared on a skip-a-day feeding program. Pullets were weighed at 20 wk of age and then distributed into 30 laying pens such that each pen had a similar BW distribution. Each individual laying pen consisted of 35 hens and 4 roosters. At 21 wk of age, the birds were photostimulated for reproduction and 15 of the laying pens were placed on a once-a-day feeding schedule, whereas the other 15 pens were placed on a twice-a-day feeding schedule. The total amount of feed provided per day to all the laying pens was the same. Birds fed once a day received all their feed at 0630 h, whereas birds fed twice a day received 60% of their total feed allotment at 0630 h and the other 40% at 1500 h. Even though both treatment groups began egg production at the end of wk 23, birds fed twice a day laid more (P < or = 0.05) eggs through 42 wk of age than those fed once a day. Additionally, the average egg weight for the entire production period, which lasted until the birds were 60 wk of age, was greater for hens fed twice a day. Overall BW uniformity for the entire experiment was significantly better for hens fed twice a day vs. once a day. However, cumulative mortality was significantly higher for hens fed twice a day than for those fed once a day. The results indicate that feeding broiler breeder hens twice a day after photostimulation may enhance reproductive performance during the early lay period.
Assuntos
Criação de Animais Domésticos/métodos , Galinhas/fisiologia , Ingestão de Alimentos/fisiologia , Oviposição/fisiologia , Estimulação Luminosa , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Galinhas/crescimento & desenvolvimento , Ovos/análise , Ovos/normas , Feminino , Distribuição Aleatória , Reprodução/fisiologia , Fatores de Tempo , Aumento de PesoRESUMO
Increased dietary protein intake rapidly (3 h) decreases malic enzyme and increases hepatic histidase mRNA expression. Experiments were conducted to determine the role that individual dispensable amino acids and nonprotein N sources might have in regulating the activity of these enzymes and to determine if the addition of a N supplement to a practical broiler diet during the entire rearing period would reduce abdominal fat accumulation in broilers. Broiler chicks were fed a basal diet containing 22% protein or this diet supplemented with 9.5% l-Glu, 5% Gly, 6% l-Ala, 5.08% ammonium bicarbonate, or 4.25% dibasic ammonium phosphate for 24 h. Each of the dietary supplements added 0.90% total N to the diet. Hepatic malic enzyme mRNA expression was significantly (P < 0.05) depressed in chicks fed any of the supplemented diets compared with chicks fed the basal diet. Histidase mRNA expression, however, was only significantly increased in the chicks fed the basal diet supplemented with Gly. Broilers fed practical corn-soybean meal starter and developer diets supplemented with 2.3, 4.7, or 9.5% Glu from 0 to 40 d of age had significantly smaller abdominal fat pads relative to BW than broilers fed the unsupplemented corn-soybean meal diets. Feeding the Glu supplements, however, reduced the overall BW gain of broilers by 100 to 150 g compared with broilers fed the unsupplemented diets. The results suggest that hepatic mRNA expression of malic enzyme may be regulated by total dietary N intake, whereas hepatic mRNA expression of histidase may be regulated by specific amino acids.
Assuntos
Galinhas/genética , Dieta/veterinária , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Fígado/enzimologia , Malato Desidrogenase/genética , Nitrogênio/farmacologia , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Comportamento Alimentar , Histidina Amônia-Liase/genética , Nitrogênio/administração & dosagem , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
1. Since high concentrations of zinc are associated with reduced crop yields, environmental concerns are emerging regarding zinc accumulation in areas where poultry production is prevalent. This study investigates growth performance and zinc utilisation during the life cycle of broilers when diets were supplemented with various concentrations of zinc from two different sources. 2. A total of 740 Cobb 500 1-d-old male broiler chicks was randomly distributed into 88 battery cages. Excreta were collected over a 48-h period on d 10, 17, 24, 31, 38, and 45 to measure zinc excretion on a dry matter basis. 3. During the 45-d experimental period, broilers were given a maize-soybean meal basal diet (30 mg/kg zinc) supplemented with 0, 20, 40, and 80 mg/kg of zinc from zinc sulphate, Availa Zn amino acid complex, or an equal combination of zinc sulphate and Availa Zn amino acid complex. 4. Progressive additions of zinc to the basal diet significantly increased body weight gain of broilers, but did not affect feed conversion or mortality. Optimum body weight gain was achieved at 80 mg/kg supplemental zinc, which exceeds the NRC recommendations of 40 mg/kg. 5. During each excreta collection period, increased supplemental zinc concentrations significantly increased zinc excretion. However, the zinc source did not influence zinc utilisation. 6. As compared with zinc sulphate supplementation, the cumulative zinc excretion data indicate that adding zinc to the basal diet in the form of Availa Zn amino acid complex decreased zinc excretion.
Assuntos
Ração Animal , Galinhas/crescimento & desenvolvimento , Dieta , Suplementos Nutricionais , Zinco/farmacologia , Animais , Meio Ambiente , Fezes , Masculino , Glycine max , Aumento de Peso/efeitos dos fármacos , Zea mays , Sulfato de Zinco/farmacologiaRESUMO
1. Environmental accumulation of zinc and copper is becoming a concern in areas having intensive animal production. This study examined performance and excretion of broiler chicks given diets supplemented with graded concentrations of zinc and copper from three different sources. 2. Two experiments were conducted, each utilising 570 1-d-old chicks. In Experiment 1, chicks were given diets containing supplemental zinc concentrations from 40 to 120 mg/kg supplied as zinc sulphate, a zinc amino acid complex (Availa Zn), or a combination of zinc sulphate and Availa Zn with each contributing one-half of the total supplemental zinc. In Experiment 2, broiler chicks were given diets with graded concentrations of supplemental copper ranging from 4 to 12 mg/kg from copper sulphate, a copper amino acid complex (Availa Cu), or a combination of copper sulphate and Availa Cu with each contributing one-half of the total supplemental copper. 3. During the 17-d experimental period, mineral concentration or source did not influence body weight, feed conversion, or the incidence of mortality from 1 to 17 d. 4. Decreasing dietary zinc concentration from 120 to 40 mg/kg reduced zinc excretion by 50%. 5. Copper excretion was decreased by 35% as supplemental copper was reduced from 12 to 4 mg/kg. 6. Reducing dietary zinc and copper concentrations can potentially decrease the accumulation of heavy metals in the environment without compromising bird performance.
Assuntos
Galinhas/crescimento & desenvolvimento , Cobre/farmacologia , Dieta , Suplementos Nutricionais , Zinco/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Cobre/administração & dosagem , Cobre/deficiência , Digestão , Meio Ambiente , Metais Pesados/farmacocinética , Zinco/administração & dosagem , Zinco/deficiênciaRESUMO
Hypothalamic neuroendocrine dopaminergic neurons exhibit a diurnal rhythm. Higher level input to these neurons has not been described. In the present study, we identified fibers known to originate in the suprachiasmatic nucleus (SCN), which were associated with neuroendocrine dopaminergic neurons. Hypothalamic sections were obtained from either ovariectomized (OVX) female rats or OVX female rats implanted with estrogen and progesterone (E+P). Confocal microscopic images were acquired from the periventricular nucleus, as well as the rostral, dorsomedial, ventrolateral, and caudal regions of the arcuate nucleus. Using antibodies directed against vasoactive intestinal peptide (VIP) and tyrosine hydroxylase (TH) the rate-limiting enzyme in dopamine synthesis, fine VIP fibers in close apposition to TH-immunoreactive (IR) soma and proximal dendrites were revealed. Of the antibodies for the two VIP receptor subtypes (VIP1R and VIP2R), only VIP2R was found on TH-IR neurons. E+P significantly increased the incidence and density of neuroendocrine dopaminergic neurons expressing VIP2R, when compared to OVX animals. E+P did not affect the percent of neuroendocrine dopaminergic neurons associated with VIP fibers. No VIP fibers or VIP2R were found on dopaminergic neurons in the zona incerta. Brain sections triple labeled for Synapsin (a protein localized in synaptic vesicles) VIP, and TH demonstrated that Synapsin was colocalized with VIP fibers that were associated with TH-IR neurons in the arcuate nucleus. Double-label immuno-electron microscopy of hypothalamic sections labeled with antibodies for VIP and TH revealed VIP boutons associated with TH-IR soma and proximal dendrites. These results suggest VIPergic neurons may directly regulate neuroendocrine dopaminergic neuron activity, and ovarian steroids may play a modulatory role.
Assuntos
Dopamina/fisiologia , Fibras Nervosas/fisiologia , Peptídeo Intestinal Vasoativo/farmacologia , Animais , Encéfalo/metabolismo , Encéfalo/fisiologia , Dendritos/metabolismo , Dendritos/fisiologia , Feminino , Imuno-Histoquímica , Neurônios/metabolismo , Neurônios/fisiologia , Sistemas Neurossecretores/metabolismo , Sistemas Neurossecretores/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores de Peptídeo Intestinal Vasoativo/biossíntese , Coloração e Rotulagem , Núcleo Supraquiasmático/metabolismo , Núcleo Supraquiasmático/fisiologia , Peptídeo Intestinal Vasoativo/metabolismoRESUMO
The isolation and identification of a prolactin-releasing factor (PRF) from the neuro-intermediate lobe of the pituitary gland has been pursued for over a decade. Using high-pressure liquid chromatography with electrochemical detection (HPLC-ECD) and gas chromatography/mass spectrometry (GC/MS) (R)-salsolinol (SAL) (a dopamine-related stereo-specific tetrahydroisoquinoline) was found to be present in neuro-intermediate lobe as well as median eminence extracts of male, intact-, and ovariectomized female rats. Moreover, analysis of SAL concentrations in neuro-intermediate lobe revealed parallel increases with plasma prolactin in lactating rats exposed to a brief (10 min) suckling stimulus following 4-h separation. SAL appears to be a selective and potent stimulator of prolactin secretion in vivo and it was without effect on the secretion of other pituitary hormones. We have also found that SAL can elevate prolactin release, although to a lesser extent, in pituitary cell cultures as well as in hypophysectomized rats bearing anterior lobe transplants under the kidney capsule. Lack of interference of SAL with [3H]-spiperone binding to AP homogenates indicates that SAL does not act at the dopamine D2 receptor. Moreover, [3H]-SAL binds specifically to homogenate of AL as well as neuro-intermediate lobe obtained from lactating rats. Taken together, our data clearly suggest that SAL is synthesized in situ and this compound can play a role in the regulation of pituitary prolactin secretion.
Assuntos
Isoquinolinas/metabolismo , Neuro-Hipófise/metabolismo , Prolactina/metabolismo , Animais , Sítios de Ligação , Feminino , Isoquinolinas/isolamento & purificação , Isoquinolinas/farmacologia , Masculino , Ovariectomia , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Neuro-Hipófise/química , Neuro-Hipófise/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Valores de Referência , Extratos de Tecidos/químicaRESUMO
Ample evidence indicates feedback relationships between pituitary prolactin and hypothalamic dopaminergic neurons. Since the presence of prolactin receptors was earlier demonstrated in hypothalamic dopaminergic neurons, our working hypothesis was that prolactin induced activation of prolactin receptor coupled signaling leads to increased neuronal activity in these neurons. The aim of this study was to correlate prolactin receptor mediated signaling and prolactin induced activation in hypothalamic dopaminergic neurons. We used nuclear translocation of STAT5 as a marker of prolactin receptor induced signaling and expression of Fos related antigens (FRAs) as an indicator of neuronal activation. We performed double label immunocytochemical studies to determine the time course of the presence of FRAs and STAT5 in the nuclei of hypothalamic dopaminergic neurons after ovine prolactin treatment. Exogenous ovine prolactin treatment of ovariectomized rats resulted in an increase in serum ovine prolactin levels and a decrease in endogenous serum prolactin levels, indicating that ovine prolactin activated mechanisms inhibited pituitary prolactin secretion. Indeed, ovine prolactin activated the prolactin receptors in most subpopulations of hypothalamic dopaminergic neurons, resulting in nuclear translocation of STAT5. Also, increased neuronal activity, indicated by expression of FRAs, was observed in the same neuron populations after ovine prolactin treatment. These results suggest that signal transduction mechanisms coupled to prolactin receptors in hypothalamic dopaminergic neurons resemble those observed in other tissues; and nuclear translocation of STAT5 can be used as a marker of prolactin receptor activation in hypothalamic dopaminergic neurons.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Dopamina/metabolismo , Hipotálamo/metabolismo , Proteínas do Leite , Neurônios/metabolismo , Prolactina/sangue , Proteínas Proto-Oncogênicas c-fos/biossíntese , Transativadores/metabolismo , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Feminino , Hipotálamo/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Prolactina/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores da Prolactina/metabolismo , Fator de Transcrição STAT5 , OvinosRESUMO
It has been previously found that all hormone-producing phenotypes of the anterior lobe of the pituitary gland are capable of producing endothelin (ET)-like substances. The aim of this study was to determine whether the expression of ET-1-like peptides in lactotrophs, gonadotrophs, and somatotrophs is influenced by different in vivo ovarian hormonal conditions. Anterior lobes of the pituitary gland were harvested from ovariectomized and ovarian steroid-replaced adult female rats 10-12 d after surgery. Quantitative immunocytochemistry was performed on enzymatically dispersed pituitary cells. The presence of ET-1-like immunoreactivity in prolactin-, luteinizing hormone-, or growth hormone-producing cells was demonstrated by double-label immunocytochemistry. The incidence of ET-1 immunopositive pituitary cells was unaffected by progesterone treatment alone. Estradiol replacement caused a modest decrease in the number of lactotrophs and somatotrophs expressing ET-1 but increased the incidence of ET-1 immunopositive cells among gonadotrophs. Combined treatment with estradiol and progesterone robustly increased the incidence of ET-1 immunopositive lactotrophs and gonadotrophs but had no effect on somatotrophs. These data reveal that the synthesis of ET-1-like peptides in lactotrophs and gonadotrophs (and, to a lesser extent, in somatotrophs) is sensitive to ovarian steroids. Furthermore, these findings predict that ovarian steroids modulate ET-1 biosynthesis during the estrous cycle, suggesting a possible mechanism by which the ovarian steroid milieu may regulate the responsiveness of lactotrophs and gonadotrophs to their hypothalamic secretagogues.
Assuntos
Endotelina-1/análise , Estradiol/farmacologia , Hormônio do Crescimento/análise , Adeno-Hipófise/química , Progesterona/farmacologia , Prolactina/análise , Animais , Implantes de Medicamento , Estradiol/administração & dosagem , Feminino , Imunofluorescência , Hormônio Foliculoestimulante/análise , Imuno-Histoquímica , Hormônio Luteinizante/análise , Ovariectomia , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Progesterona/administração & dosagem , Ratos , Ratos Sprague-DawleyRESUMO
The term paraesophageal hernia is described as a herniation of the gastric fundus through the open hiatus into the thoracic cavity while the lower esophageal sphincter (LES) remains in its normal anatomic position. This is considered a rolling esophageal hernia (Type II), and it is the least commonly encountered hiatal hernia. A more commonly encountered herniation of the fundus of the stomach is the Type III hernia, in which both the LES and the fundus herniate into the chest. This has also been classified as a paraesophageal hernia. The most common hiatal hernia is a sliding hiatal hernia (Type I), which consists of herniation of the stomach through the esophageal hiatus, causing the LES and gastric cardia to lie in the thoracic cavity. There are several controversial issues involved in paraesophageal hernia repair, including indications for surgery, the most appropriate surgical approach, and the need for a concomitant antireflux procedure. The increasing popularity of laparoscopic paraesophageal hernia repair has dramatically altered the approach to these patients and has allowed patients at higher risk to better tolerate this procedure with a decrease in morbidity and mortality. However, they remain difficult surgical procedures.
Assuntos
Hérnia Hiatal/cirurgia , Laparoscopia , HumanosRESUMO
We have previously found that lactotrophs express and secrete endothelin-like peptides that influence prolactin (PRL) secretion in an autocrine fashion. We have also observed that the incidence of endothelin-immunoreactive lactotrophs is markedly affected by ovarian steroids. In this study, we examined how the ovarian steroid background determines the efficiency of the endothelin-mediated autocrine feedback regulation of PRL secretion. Ovariectomized adult female rats were used throughout these studies. Steroid replacements were made by sc implantation of Silastic capsules immediately following ovariectomy. Eight to 10 wk later, three animals from each treatment group (no steroid control, estradiol, progesterone, estradiol plus progesterone) were sacrificed by decapitation, and the anterior pituitary cells were enzymatically dispersed using collagenase and hyaluronidase. A PRL-specific reverse hemolytic plaque assay was used to measure PRL secretion at the single-cell level. BQ123, a synthetic cyclic pentapeptide with distinctive endothelin-A receptor antagonist quality, caused only a modest elevation of PRL secretion in the control group. Endothelin antagonism did not affect PRL secretion in cells obtained from progesterone-implanted animals. Endothelin antagonism did, however, increase overall PRL secretion in the estradiol and estradiol plus progesterone groups by five- and threefold, respectively. Frequency distribution of PRL plaques in these same two BQ123-treated groups revealed two subpopulations, indicating that lactotrophs differ in their response to endogenous endothelin feedback and that this difference is steroid dependent. These observations clearly suggest that the ovarian steroid milieu (estrogens in particular) can have a profound influence on the self-regulatory mechanisms of lactotrophs. Our results also emphasize that endogenous endothelins may play an important role in the negative feedback regulation of PRL secretion in female rats.
Assuntos
Comunicação Autócrina , Endotelinas/fisiologia , Prolactina/metabolismo , Animais , Combinação de Medicamentos , Antagonistas dos Receptores de Endotelina , Estradiol/farmacologia , Retroalimentação , Feminino , Ovariectomia , Peptídeos Cíclicos/farmacologia , Adeno-Hipófise/citologia , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Progesterona/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
In our previous studies we found that administration of exogenous prolactin increased dopamine turnover in the terminal areas of the hypothalamic dopaminergic neurons controlling prolactin secretion from pituitary lactotrophs. In this study we investigated the effect of immunoneutralization of endogenous prolactin on the expression of FRAs in the tuberoinfundibular dopaminergic (TIDA), tuberohypophysial dopaminergic (THDA), and periventricular hypothalamic dopaminergic (PHDA) subpopulations of the hypothalamic dopaminergic neurons. Female rats were ovariectomized on d 0 of the experiment. At 1000 h of d 10, all animals were injected with 20 microg of 17-beta-estradiol sc to induce a proestrous-like surge of prolactin at 1700 h the next day. At 1000 h on d 11, half of the animals were injected with 200 microL of rabbit anti-rat prolactin antiserum ip, while the controls received normal rabbit serum. Groups of animals were sacrificed for immunocytochemistry in 2 h intervals between 1300 and 2100 h. Double-label immunocytochemistry for FRAs and tyrosine hydroxylase (TH) was performed and the results are presented as percentage of TH-immunoreactive neurons expressing FRAs. In the control animals, expression of FRAs decreased at 1500 h, gradually increased by 1900 h, but was lower than the basal levels by 2100 h. Expression of FRAs was significantly lower at 1900 h in the PHDA, THDA and TIDA neurons of prolactin antiserum treated rats than in the controls. These results indicate that elimination of endogenous prolactin from the circulation lowers the activity and/or prevents the reactivation of neuroendocrine dopaminergic neurons at the beginning of the dark phase.
Assuntos
Dopamina/metabolismo , Hipotálamo/metabolismo , Soros Imunes/farmacologia , Prolactina/imunologia , Proteínas Proto-Oncogênicas c-fos/análise , Animais , Núcleo Arqueado do Hipotálamo/química , Estradiol/farmacologia , Feminino , Hipotálamo/química , Imuno-Histoquímica , Neurônios/química , Neurônios/metabolismo , Ovariectomia , Prolactina/sangue , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Distribuição Tecidual , Tirosina 3-Mono-Oxigenase/análiseRESUMO
Prolactin is a protein hormone of the anterior pituitary gland that was originally named for its ability to promote lactation in response to the suckling stimulus of hungry young mammals. We now know that prolactin is not as simple as originally described. Indeed, chemically, prolactin appears in a multiplicity of posttranslational forms ranging from size variants to chemical modifications such as phosphorylation or glycosylation. It is not only synthesized in the pituitary gland, as originally described, but also within the central nervous system, the immune system, the uterus and its associated tissues of conception, and even the mammary gland itself. Moreover, its biological actions are not limited solely to reproduction because it has been shown to control a variety of behaviors and even play a role in homeostasis. Prolactin-releasing stimuli not only include the nursing stimulus, but light, audition, olfaction, and stress can serve a stimulatory role. Finally, although it is well known that dopamine of hypothalamic origin provides inhibitory control over the secretion of prolactin, other factors within the brain, pituitary gland, and peripheral organs have been shown to inhibit or stimulate prolactin secretion as well. It is the purpose of this review to provide a comprehensive survey of our current understanding of prolactin's function and its regulation and to expose some of the controversies still existing.
Assuntos
Prolactina/metabolismo , Processamento Alternativo/genética , Animais , Mama/efeitos dos fármacos , Mama/metabolismo , Linhagem Celular , Ritmo Circadiano/fisiologia , Feminino , Homeostase/efeitos dos fármacos , Homeostase/fisiologia , Humanos , Linfócitos/metabolismo , Especificidade de Órgãos/efeitos dos fármacos , Especificidade de Órgãos/fisiologia , Adeno-Hipófise/metabolismo , Gravidez , Prolactina/genética , Prolactina/farmacologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/farmacologia , Receptores da Prolactina/genética , Receptores da Prolactina/metabolismo , Reprodução/efeitos dos fármacos , Reprodução/fisiologia , Transdução de Sinais/fisiologia , Relação Estrutura-AtividadeRESUMO
The secretion of prolactin (PRL) from the anterior lobe (AL) of the pituitary gland is tonically inhibited by dopamine (DA) of hypothalamic origin. While ovarian steroids play a role in the regulation of the secretion of PRL, their effect on all three populations of hypothalamic neuroendocrine dopaminergic neurons is not fully understood. In this study we describe the effects of ovarian steroids on regulation of the release of DA from tuberoinfundibular dopaminergic (TIDA), tuberohypophyseal dopaminergic (THDA) and periventricular-hypophyseal dopaminergic (PHDA) neurons. Adult female rats were bilaterally ovariectomized (OVX) and, 10 days following ovariectomy (day 0), injected with corn oil (vehicle), estrogen, or estrogen plus progesterone (day 1). Animals were sacrificed every 2 h from 09.00 to 21.00 h by rapid decapitation. Trunk blood was collected and the concentration of PRL in serum was determined by radioimmunoassay. The median eminence (ME) and the AL, intermediate (IL) and neural (NL) lobes of the pituitary gland were dissected and the concentration of DA and DOPAC in each was measured by HPLC-EC. OVX rats presented small but significant increases in the secretion of PRL at 15.00 and 17.00 h. Replacement of estrogen or estrogen plus progesterone increased the basal concentration of PRL. Moreover, injection of estrogen only, or estrogen plus progesterone increased the concentration of PRL in serum at 15.00 h through 19.00 h, respectively, followed by a decrease to baseline thereafter. The turnover of DA in the ME and NL of OVX rats increased at 13.00 and returned to low levels. Turnover of DA in the IL of OVX rats increased in the morning by 11.00 h and remained elevated before decreasing by 17.00 h. The turnover of DA in the ME, IL and NL of OVX rats increased by 19.00 h. Injection of estrogen advanced the increase of TIDA activity by 2 h in the ME compared to OVX rats. Moreover, administration of estrogen suppressed the activity of THDA and PHDA neurons in the afternoon compared to OVX rats. In estrogen plus progesterone-treated rats, the activity of hypothalamic neuroendocrine dopaminergic neurons terminating in the ME, IL, and NL was inhibited prior to the increase in the secretion of PRL. The concentration of DA in the AL diminished prior to the estrogen-induced increase of PRL. Administration of progesterone, in concert with estrogen, delayed the increase of PRL in serum and the decrease of DA in the AL, compared to estrogen-treated rats, by 4 h. These data suggest a major role for ovarian steroids in controlling increases in the secretion of PRL by not only stimulating PRL release from lactotrophs, but also by inhibiting the activity of all three populations of hypothalamic neuroendocrine DAergic neurons.
Assuntos
Ritmo Circadiano , Dopamina/metabolismo , Estrogênios/farmacologia , Eminência Mediana/fisiologia , Neurônios/fisiologia , Hipófise/fisiologia , Progesterona/sangue , Progesterona/farmacologia , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Animais , Feminino , Eminência Mediana/efeitos dos fármacos , Ovariectomia , Hipófise/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
We have found that exogenous prolactin (PRL) stimulates all three populations of hypothalamic neuroendocrine dopaminergic neurons. In this study, we investigated the effects of immunoneutralization of endogenous PRL on the activity of these neurons. Injection of 17beta-estradiol (E2) (20 microg subcutaneously) 10 d after ovariectomy induced a proestrus-like increase in PRL in peripheral plasma the following afternoon. At 1000 h the day after E2 injection, rats received either rabbit antirat PRL antiserum (PRL-AS) (200 microL) or normal rabbit serum (NRS, 200 microL, controls) intraperitoneally. Groups of rats were then decapitated every 2 h from 1100 h to 2100 h. Trunk blood was collected and serum extracted with protein A to remove the PRL-AS/PRL complex, and the remaining free PRL was measured by radioimmunoassay. Sites of neuroendocrine dopaminergic nerve terminals, the median eminence (ME), and intermediate and neural lobes of the pituitary gland were excised and stored for determination of dopamine (DA) and 3,4-dihydroxyphenyl acetic acid (DOPAC) concentrations by high-performance liquid chromatography electrochemical detection (EC). In addition, the anterior lobe of the pituitary gland, the locus of DA action, was collected. The concentration of PRL in NRS-treated animals increased by 1500 h, peaked by 1700 h, and returned to low levels by 2100 h. PRL-AS prevented the increase in PRL secretion in response to E2. The turnover of DA (DOPAC:DA ratio; an index of dopaminergic neuronal activity) in the ME of NRS-treated animals increased at 1500 h and rapidly returned to basal levels. Treatment with PRL-AS prevented the increase in DA turnover in the ME. DA turnover in the intermediate lobe increased coincident with the peak of PRL in serum of NRS-treated rats. PRL-AS administration prevented increased DA turnover in the intermediate lobe. The turnover of DA in the neural lobe increased by 1300 h and decreased steadily through 2100 h. However, administration of PRL-AS minimally suppressed the turnover of DA in the neural lobe. Moreover, administration of PRL-AS attenuated the rise of DA in the anterior lobe associated with the waning phase of the E2-induced PRL surge. These results clearly indicate that endogenous PRL regulates its own secretion by activating hypothalamic neuroendocrine dopaminergic neurons.
Assuntos
Dopamina/fisiologia , Hipotálamo/efeitos dos fármacos , Soros Imunes/farmacologia , Neurônios/efeitos dos fármacos , Prolactina/farmacologia , Prolactina/fisiologia , Ácido 3,4-Di-Hidroxifenilacético/análise , Animais , Cromatografia Líquida de Alta Pressão , Dopamina/análise , Estradiol/farmacologia , Retroalimentação , Feminino , Eminência Mediana/química , Terminações Nervosas/química , Ovariectomia , Hipófise/química , Prolactina/imunologia , Ratos , Ratos Sprague-DawleyRESUMO
Three populations of hypothalamic neuroendocrine dopaminergic (NEDA) neurons, arising from the arcuate and periventricular nuclei of the hypothalamus release dopamine (DA) that acts at the pituitary gland to regulate the secretion of PRL. It is generally accepted that NEDA neurons lack functional DA transporters (DATs), which are responsible for uptake of DA from the synaptic cleft into the presynaptic axon terminal. This study localized DATs to the hypothalamo-pituitary axis and evaluated the effect of DAT blockade on the hypothalamo-pituitary regulation of PRL. After 7 days of treatment with cocaine (a nonspecific amine transporter blocker) or mazindol (a specific DAT blocker), the relative abundance of PRL messenger RNA (mRNA) in the anterior lobe (AL) of OVX rats was significantly decreased, whereas the relative abundance of tyrosine hydroxylase mRNA in the hypothalamus was significantly increased. The effect of cocaine or mazindol administration on DA turnover and serum PRL concentration was examined in estradiol (E2)-treated OVX rats. E2 administration (i.v.) resulted in a significant increase in serum PRL within 4 h; however, cocaine or mazindol administration abolished the E2-induced increase of PRL. Cocaine or mazindol significantly increased the concentration of DA at the site of the axon terminals within the median eminence (ME), intermediate lobe (IL) and neural lobe (NL), indicating blockade of uptake. Because formation of DOPAC requires uptake of DA, concentrations of DOPAC in the ME, IL and NL decreased following treatment with either cocaine or mazindol. These data, together with the presence of immunopositive DAT in the ME, pituitary stalk, IL, and NL, suggest that a functional DAT system is present within all three populations of NEDA neurons. Moreover, similarity between the effects of cocaine and mazindol treatment indicate that blockade of the DAT, but not other amine transporters, is responsible for suppression of PRL gene expression and secretion. Blockade of DATs prevent uptake of DA into NEDA neurons and consequently increases the amount of DA that diffuses into the portal vasculature and reaches the AL. These data provide evidence that DATs play a physiological role in the regulation of DA release from and TH expression in NEDA neurons and consequently PRL secretion and PRL gene expression and further support our previous observation that the regulation of PRL secretion involves all three populations of NEDA neurons.
Assuntos
Proteínas de Transporte/fisiologia , Dopamina/fisiologia , Glicoproteínas de Membrana , Proteínas de Membrana Transportadoras , Proteínas do Tecido Nervoso/fisiologia , Prolactina/metabolismo , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Cocaína/farmacologia , Dopamina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina , Inibidores da Captação de Dopamina/farmacologia , Estradiol/farmacologia , Feminino , Sistema Hipotálamo-Hipofisário/fisiologia , Imuno-Histoquímica , Mazindol/farmacologia , Eminência Mediana/metabolismo , Sistemas Neurossecretores/citologia , Sistemas Neurossecretores/metabolismo , Prolactina/biossíntese , Ratos , Ratos Sprague-Dawley , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Ablation of a-calcitonin gene-related polypeptide (CGRP) containing neurons with the afferent neurotoxin capsaicin improves postoperative foregut transit in a rodent model. Similarly, administration of a selective alpha-CGRP antibody or hCGRP((8-37)), a CGRP receptor antagonist, improves postoperative gastric emptying. Unlike the stomach, which contains only alpha-CGRP, the small bowel additionally contains beta-CGRP. The role of the latter in postoperative small bowel transit is unknown. The purpose of this study was to evaluate the effect of an alpha-CGRP antibody and hCGRP((8-37)) on postoperative small bowel transit. Male Sprague-Dawley rats underwent placement of duodenal catheters and were randomly assigned to 1 of 11 groups. Four groups were pretreated with 1% capsaicin. One week later, all animals underwent standardized laparotomy following administration of a control antibody or the alpha-CGRP mono-clonal antibody, or during infusion of hCGRP((8-37)) at varying doses. Small bowel transit was measured 25 minutes postoperatively. The alpha-CGRP antibody sped postoperative transit when given alone or in combination with capsaicin. In contrast, animals treated with hCGRP((8-37)) showed no significant improvement in postoperative transit, and the beneficial effect of capsaicin was blocked. Unlike their similar effects on postoperative gastric emptying, we found that hCGRP((8-37)) and the alpha-CGRP antibody had differing effects on postoperative small bowel transit. The reason for this is unknown but may be related to their differing specificities for alpha- and beta-CGRP.