RESUMO
Elevated oestradiol concentrations during the follicular phase stimulate a surge in gonadotropin-releasing hormone (GnRH) and luteinising hormone (LH) concentrations, which leads to ovulation. Progesterone can block the oestradiol-induced GnRH/LH surge, but the mechanism that is involved is unclear. We examined the effect of progesterone on oestradiol-induced activation of cells within the ovine hypothalamus/preoptic area (POA) to determine: (i) in which regions progesterone acts to block the GnRH/LH surge and (ii) whether progesterone directly or indirectly prevents activation of oestradiol-responsive cells. Cellular activation was assessed by measuring the number of cells that expressed Fos (an immediate early gene). Exposure to increased oestradiol concentrations in the absence of progesterone (which normally stimulates a LH surge) did not cause any region-specific changes in hypothalamic Fos expression during the activation stage of the LH surge-induction process (Experiment 1). The same treatment significantly increased cellular activation within the POA, lateral septum (LS), and arcuate nucleus at the time of surge onset (Experiment 2). Concurrent exposure to increased oestradiol and progesterone concentrations during the activation stage of the surge-induction process (which normally blocks the LH surge) was associated with significantly reduced cellular activation within the ventromedial hypothalamus and anterior hypothalamic area, relative to the positive controls (oestradiol increment alone) and arcuate nucleus relative to the negative controls (no increment in oestradiol) during the activation stage (Experiment 1). At the time of surge onset (Experiment 2), exposure to progesterone during the activation period prevented the oestradiol-induced increase in cellular activation that occurred in the POA, LS and arcuate nucleus of the positive controls. These results demonstrated that oestradiol and progesterone induced differential region- and time-specific effects on cellular activation within the regions of the ovine brain that generate the preovulatory GnRH/LH surge. Moreover, the lack of cellular activation within the POA, LS and arcuate nucleus at the time of surge onset in animals exposed to progesterone during the activation stage is consistent with the hypothesis that progesterone can block the preovulatory surge by direct inhibition of oestradiol-induced cellular activation in these areas.
Assuntos
Estradiol/metabolismo , Ciclo Estral/sangue , Hormônio Liberador de Gonadotropina/sangue , Hipotálamo/metabolismo , Hormônio Luteinizante/sangue , Progesterona/fisiologia , Animais , Núcleo Arqueado do Hipotálamo/citologia , Núcleo Arqueado do Hipotálamo/metabolismo , Regulação para Baixo , Feminino , Hipotálamo/citologia , Sistemas Neurossecretores/fisiologia , Área Pré-Óptica/citologia , Área Pré-Óptica/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Septo do Cérebro/citologia , Septo do Cérebro/metabolismo , OvinosRESUMO
Puberty is triggered by an increase in pulsatile release of luteinizing hormone-releasing hormone (LHRH) from the hypothalamus. Although the LHRH neurosecretory system is mature well before the onset of puberty, a central inhibitory mechanism restrains LHRH release in juvenile primates. Recent studies suggest that this central inhibition is primarily because of GABAergic neurotransmission. A reduction of GABAergic restraint appears to be essential for the initiation of puberty, but the mechanism that underlies the disinhibition process remains to be elucidated. Future research into the regulation of central inhibition should provide more effective treatments for the prevention of disease associated with abnormal pubertal development.
Assuntos
Encéfalo/fisiologia , Hormônio Liberador de Gonadotropina/metabolismo , Macaca mulatta/fisiologia , Maturidade Sexual/fisiologia , Envelhecimento , Animais , Encéfalo/crescimento & desenvolvimento , Feminino , Ácido Glutâmico/fisiologia , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/fisiologia , Masculino , Ácido gama-Aminobutírico/fisiologiaRESUMO
How progesterone blocks the E2-induced GnRH surge in females is not known. In this study we assessed whether the endogenous opioid peptides (EOPs) that mediate progesterone negative feedback on pulsatile GnRH secretion also mediate the blockade of the GnRH surge. We treated ovariectomized ewes with physiological levels of E2 and progesterone to stimulate and block the GnRH surge, respectively, using LH secretion as an index of GnRH release. A pilot study confirmed that blocking opioidergic neurotransmission with the opioid receptor antagonist, naloxone (NAL; 1 mg/kg.h, i.v.), could prevent the suppression of pulsatile LH secretion by progesterone in our model. By contrast, antagonizing EOP receptors with NAL did not restore LH surges in ewes in which the E2-induced GnRH surge was blocked by progesterone treatment during the E2-dependent activation stage (Exp 1) of the GnRH surge induction process. However, in ewes treated with progesterone during the E2-independent transmission stage (Exp 2), NAL partially restored blocked LH surges, as indicated by increased fluctuations in LH that, in some cases, resembled LH surges. We conclude, therefore, that the EOPs that mediate progesterone negative feedback on pulsatile GnRH secretion are not involved in blockade of activation of the E2-induced GnRH surge by progesterone, but do appear to be part of the mechanism by which progesterone disrupts the transmission stage.
Assuntos
Endorfinas/fisiologia , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Hormônio Liberador de Gonadotropina/metabolismo , Progesterona/fisiologia , Animais , Estradiol/farmacologia , Retroalimentação , Feminino , Hormônio Luteinizante/metabolismo , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Ovariectomia , Progesterona/farmacologia , OvinosRESUMO
Progesterone can either augment or inhibit the surge of gonadotropin-releasing hormone (GnRH) that drives the preovulatory luteinizing hormone (LH) surge. This study investigated the central mechanisms through which progesterone might achieve these divergent effects by examining the effects of exogenous steroids on the activation of GnRH neurons and non-GnRH-immunopositive cells in the preoptic area/anterior hypothalamus of steroid-treated ovariectomized ewes. Fos expression (an index of cellular activation) was examined during the estradiol-induced GnRH surge in ewes treated with progesterone using regimes that have been reported to either augment (progesterone pretreatment) or inhibit (progesterone treatment at the time of the surge-inducing estradiol increment) the GnRH surge. Control groups received either no progesterone pretreatment or no surge-inducing estradiol increment. Induction of an LH surge was associated with a significant (p < 0.0001) increase in the proportion of activated GnRH neurons, irrespective of whether ewes received progesterone pretreatment. However, the number of non-GnRH-immunopositive cells activated during the surge was significantly (p < 0.0001) increased in ewes that received the progesterone pretreatment. By contrast, the proportion of GnRH neurons and non-GnRH-immunopositive cells that expressed Fos was significantly (p < 0.0001) reduced in ewes in which the surge was inhibited by progesterone compared to ewes in which a surge was stimulated. These data indicate that (1) progesterone pretreatment increases the activation of non-GnRH cells during the estradiol-induced surge, but does not affect the proportion of GnRH neurons activated and (2) when administered concurrently with a surge-inducing estradiol increment, progesterone prevents the activation of GnRH neurons and non-GnRH cells that is normally associated with the estradiol-induced surge. Therefore, progesterone does not appear to augment the GnRH surge by increasing the proportion of GnRH neurons that are activated by estradiol, whereas inhibition of the GnRH surge involves prevention of the activation of GnRH neurons. Thus, the augmentation and inhibition of the GnRH surge by progesterone appear to be regulated via different effects on the GnRH neurosecretory system.
Assuntos
Estradiol/farmacologia , Hormônio Liberador de Gonadotropina/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Progesterona/farmacologia , Animais , Feminino , Hormônio Liberador de Gonadotropina/análise , Hipotálamo Anterior/química , Imuno-Histoquímica , Hormônio Luteinizante/metabolismo , Neurônios/química , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Ovariectomia , Área Pré-Óptica/química , Área Pré-Óptica/efeitos dos fármacos , Área Pré-Óptica/fisiologia , Proteínas Proto-Oncogênicas c-fos/análise , OvinosRESUMO
The preovulatory surges of GnRH and LH are activated by increased concentrations of circulating estradiol, but ovulation is blocked when progesterone concentrations are elevated. Although it is has been shown that this action of progesterone is due to a central inhibition of the GnRH surge, the mechanisms that underlie the blockade of the GnRH surge are poorly understood. In this study we investigated whether progesterone can block the estradiol-dependent activation stage of the GnRH surge induction process, and thus prevent expression of the LH surge. The results demonstrated that exposure to progesterone for half or the full duration of the activation stage can prevent the stimulation of LH surges by estradiol (experiment 1), whereas exposure to progesterone midway though a period of estradiol exposure, which in itself is sufficient to activate the surge, did not block the LH surge (experiment 2). These results suggest that progesterone 1) disrupts activation of the surge induction system in response to a stimulatory estradiol signal and 2) does not compromise the ability of animals to respond to a stimulatory estradiol signal applied immediately after progesterone exposure. Because the disruptive effects of activated progesterone in response to estradiol are rapid but transient, it may be that progesterone directly interferes with the activation of estradiol-responsive neural systems to block the GnRH/LH surge.
Assuntos
Estradiol/farmacologia , Antagonistas de Estrogênios , Hormônio Luteinizante/metabolismo , Progesterona/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Feminino , Hormônio Liberador de Gonadotropina/metabolismo , Cinética , Projetos Piloto , Gravidez , OvinosRESUMO
Sour taste is elicited by acids. How taste cells transduce sour taste is controversial because acids (specifically protons) have diverse effects on cell membranes. Consequently, it is difficult to differentiate between events related to sour taste transduction per se and unrelated effects of protons. We have studied acid taste transduction in mouse taste buds using a lingual slice preparation where it is possible to measure changes in pH and [Ca2+]i simultaneously in taste cells. Focal application of citric acid or HCl to the apical tips of taste buds produced widespread acidification of the entire taste bud. Citric acid was effective at a pH of approximately 4, but HCl only at a pH of approximately 1.5. Despite acidification of the whole taste bud, only a select few taste cells exhibited Ca2+ responses. Acid-evoked Ca2+ responses were dose dependent in a range consistent with them being sour-taste responses. Cells exhibiting acid-evoked Ca2+ responses also responded to KCl depolarization. Acid-evoked Ca2+ responses were blocked by Ba2+ (2 mM) and Cd2+ (500 microM), suggesting that acid responses are generated by Ca2+ influx through depolarization-gated Ca2+ channels. Removing extracellular Ca2+ reduced acid-evoked Ca2+ responses, but depleting intracellular Ca2+ stores with thapsigargin had no effect, suggesting that acid taste responses are generated by an influx of extracellular Ca2+. Neither Cs+ (500 microM) nor amiloride (100 microM) affected acid-evoked Ca2+ responses, suggesting that neither hyperpolarization-activated cyclic nucleotide-gated cation (pacemaker) channels nor epithelial Na+ channels, respectively, transduce sour taste. Collectively, the results indicate that acids, especially weak acids, acidify the taste bud and evoke depolarization-induced Ca2+ entry into a select subset of taste cells. The primary transducer protein(s) for sour taste remain undiscovered.
Assuntos
Cálcio/metabolismo , Ácido Cítrico/farmacologia , Ácido Clorídrico/farmacologia , Prótons , Papilas Gustativas/metabolismo , Paladar/fisiologia , Animais , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos DBA , Microscopia Confocal , Papilas Gustativas/citologia , Papilas Gustativas/efeitos dos fármacos , Língua/citologia , Língua/efeitos dos fármacos , Língua/metabolismoRESUMO
Periodic release of luteinizing hormone-releasing hormone (LHRH) from the hypothalamus is essential for normal reproductive function. Pulsatile LHRH release appears to result from the synchronous activity of LHRH neurons. However, how the activity of these neurons is synchronized to release LHRH peptide in a pulsatile manner is unclear. Because there is little evidence of physical coupling among LHRH neurons in the hypothalamus, we hypothesized that the activity of LHRH neurons might be coordinated by indirect intercellular communication via intermediary (nonneural) cells rather than direct interneural coupling. In this study, we used an in vitro preparation of LHRH neurons derived from the olfactory placode of monkey embryos to assess whether nonneuronal cells, play a role in coordinating LHRH neuronal activity. We found that cultured LHRH neurons and nonneuronal cells both exhibit spontaneous oscillations in the concentration of intracellular Ca(2+) ([Ca(2+)](i)) at similar frequencies. Moreover, [Ca(2+)](i) oscillations in both types of cell were periodically synchronized. Synchronized [Ca(2+)](i) oscillations spread as intercellular Ca(2+) waves across fields of cells that included LHRH neurons and nonneuronal cells, although waves spread at a higher velocity among LHRH neurons. These results suggest that LHRH neurons and nonneuronal cells are functionally integrated and that nonneuronal cells could be involved in synchronizing the activity of the LHRH neurosecretory network.
Assuntos
Cálcio/metabolismo , Hormônio Liberador de Gonadotropina/metabolismo , Neurônios/metabolismo , Animais , Células/metabolismo , Técnicas de Cultura , Ectoderma/citologia , Ectoderma/metabolismo , Membranas Intracelulares/metabolismo , Macaca mulatta/embriologia , Condutos Olfatórios/citologia , Condutos Olfatórios/embriologia , Oscilometria , Concentração Osmolar , Fatores de TempoRESUMO
Oestradiol provides the drive to reproductive cyclicity in female mammals through its ability to stimulate the GnRH surge. In contrast, progesterone can be seen as the 'clutch and brakes' within reproductive cycles, as it can modify the response of the GnRH neurosecretory system to oestradiol. In this regard, progesterone has multiple and sometimes opposing effects on the GnRH neurosecretory system. For example, dependent upon the timing of exposure, progesterone enhances the amplitude of the oestradiol-induced LH (rats) and GnRH surge (within cerebrospinal fluid in sheep, mRNA concentrations in rats), but can also inhibit pulsatile GnRH secretion, and delay or even block expression of the surge (monkeys, rats and sheep). Investigations of the mechanisms of action of progesterone are complicated further by the fact that some of the observed effects of progesterone, such as the ability to block the oestradiol-induced surge, appear to be mediated via several different routes. Consequently, a variety of approaches are needed to advance our understanding of this fundamental reproductive neuroendocrine system. In this context, large animal neuroendocrine models have provided important information about the mechanisms of progesterone action and provide many exciting opportunities for future research.
Assuntos
Estradiol/fisiologia , Ciclo Estral/fisiologia , Hormônio Liberador de Gonadotropina/metabolismo , Hormônio Luteinizante/metabolismo , Progesterona/fisiologia , Animais , Feminino , Hormônio Liberador de Gonadotropina/líquido cefalorraquidiano , Haplorrinos , Modelos Animais , Ratos , Taxa Secretória , OvinosRESUMO
The springbok is an arid-adapted antelope inhabiting the desert and semidesert regions of southern Africa. Because it thrives in these sparsely vegetated areas, the springbok is of potential agricultural importance and the prospect of domestication has been speculated for many years. However, apart from observational studies on its breeding in the wild, suggesting it is an aseasonal breeder, little is known about the underlying reproductive endocrinology of this species. In this study, biweekly peripheral blood samples were collected from eight captive springbok ewes from October 1995 until September 1998 and analyzed for progesterone. At the start of the study, six ewes were prepubertal and cycling commenced spontaneously between November 1995 and June 1996. Cycling had already commenced in two ewes. At the end of November 1996, estrous cycles ceased abruptly in all ewes and restarted in April 1997. Cycling ceased again between December 1997 and February 1998 and restarted in June 1998 in six ewes; there was no cessation of estrous cycles in two ewes. Thus, although some individuals cycle continuously, there is a clear endocrine anestrus of between 4 and 5 mo in springbok, the timing and duration of which is synchronized between some individuals but the time of onset and cessation is variable from year to year. To ensure that the fluctuations we observed in progesterone levels were reliable indicators of changes in the estrous cycle, blood samples were collected every 6 h for 16 days in August 1998. A surge in LH secretion was observed in all ewes 55 +/- 5 h after the fall in progesterone. Progesterone levels increased again 45 +/- 8 h after the surge. A final study showed that the pattern of melatonin release in springbok exhibits a normal day/night profile, and thus photoperiodic information is transformed into an endocrine code to springbok but does not appear to affect reproduction. Rather, our data raise the possibility that the prevailing ambient temperature may influence the onset of ovarian activity in this species.
Assuntos
Antílopes/fisiologia , Ovulação , Periodicidade , Reprodução , Estações do Ano , Animais , Estro , Feminino , Hormônio Luteinizante/metabolismo , Progesterona/sangueRESUMO
The day- and night-time levels of plasma melatonin were measured in adult male and female highveld mole-rats, Cryptomys hottentotus pretoriae. This study aimed to assess whether melatonin secretion in this nocturnal, strictly subterranean but seasonally breeding rodent has a day-night rhythm and whether that rhythm is circadian and can be modified by photoperiod. In experiment 1, a day-night rhythm of plasma melatonin was found in all animals housed on a 12L:12D schedule, with significantly higher concentrations in the dark (D) compared with the light (L) phase. The increment of plasma melatonin concentration at night was the same on days 1 and 2 for animals in the control group and animals transferred to constant dark. The animals transferred to constant light substantially reduced the amplitude of the melatonin rhythm on day 2. This suggests that the endogenous melatonin rhythm in C. h. pretoriae has a circadian pattern, which can be synchronized by photoperiod and inhibited by exposure to light at night. In experiment 2, the concentration of plasma melatonin in animals kept under 14L:10D (long day, LD) conditions differed significantly from animals on 10L:14D (short day, SD). This finding supports the notion that C. h. pretoriae is sensitive to changes in day length.